Immunization with oligosaccharide conjugate Haemophilus influenzae type b (HbOC) vaccine on a large health maintenance organization population: extended follow-up and impact on Haemophilus influenzae disease epidemiology. The Kaiser Permanente Pediatric Vaccine Study Group. Between February, 1988, and June, 1990, the safety, immunogenicity and efficacy of the HbOC (oligosaccharide conjugate Haemophilus influenza type b) vaccine was evaluated in a prelicensure trial performed in a study population of 61,080 children within the Northern California Kaiser Permanente Medical Care Program. In this evaluation the HbOC vaccine was found to be safe, immunogenic and efficacious in infancy. Since licensure an estimated 162,000 additional doses of HbOC vaccine have been given to 75,000 additional children. In addition to reporting on extended follow-up of this population, this publication reports on the impact of immunizing a high proportion of the Northern California Kaiser Permanente Medical Care Program population in infancy and early childhood on the epidemiology of invasive disease caused by H. influenzae type b (Hib) and invasive disease caused by non-type b H. influenzae. As of January 31, 1992, six cases of Hib invasive disease have been identified in vaccinated children. Of these five occurred in children who had received only one dose of vaccine in infancy. One case of Hib meningitis occurred in a 3 1/2-year-old child who had received doses of HbOC at 2, 4 and 6 months of age but no further doses of any Hib vaccines. During 1991 a total of three cases of invasive disease caused by Hib were observed in children younger than 18 months of age within the Northern California Kaiser Permanente Medical Care Program. This represents a 94% reduction in disease incidence in this age group from that observed in the years 1984 to 1987.(ABSTRACT TRUNCATED AT 250 WORDS)


Respiratory syncytial virus: a comparison of diagnostic modalities. This study compared prospectively viral culture for respiratory syncytial virus (RSV) with three rapid RSV antigen detection tests: RSV EIA and TestPack RSV (TP), and Directigen RSV (DIR). Additionally two methods of specimen collection were compared: nasopharyngeal rub (RUB) and nasopharyngeal wash (WASH). True positives were defined as positive RSV viral culture or at least two positive antigen tests. One hundred ninety-eight WASH specimens obtained from children 3 years of age or younger during the 1991 RSV winter epidemic were tested for RSV antigen. Sensitivity and specificity of WASH specimens were 59.5 and 100% (culture), 86.2 and 98.2% (RSV EIA), 91 and 96.3% (TP) and 83.5 and 94.3% (DIR). Concurrently obtained RUB samples from 124 children were tested by TP and/or DIR. Sensitivity of RUB specimens was significantly lower than that of WASH specimens; 64.9% (TP) and 43.6 (DIR). These easy to perform, rapid RSV antigen tests for WASH specimens provide timely diagnosis thereby facilitating management decisions and isolation efforts.


Successful intervention in a group A meningococcal outbreak in Auckland, New Zealand. During two consecutive winter seasons (1985 and 1986) Auckland, New Zealand, experienced epidemic rates of Group A meningococcal disease, a pattern not previously recognized in New Zealand. The overall rate was 8.3/100,000/year. The highest annual rate (64.7) occurred in children 0 to 23 months of age. A city-wide vaccine campaign commencing in May, 1987, was conducted over 6 weeks among children 3 months to 13 years of age with special emphasis on reaching populations at highest risk (Maori and Pacific Island Polynesian children in certain geographic regions of Auckland). Children from 2 to 13 years of age received a single dose of monovalent Group A meningococcal vaccine. Children ages 3 to 23 months received two doses at least 1 month apart. Overall approximately 130,000 doses were delivered; coverage was approximately 90% in the single dose target group. Among the younger children approximately 89% received the primary dose. Only approximately 26% received the recommended "booster" dose. After 2 1/2 years of active surveillance (1987 to 1989) there were no cases of invasive Group A meningococcal disease in children appropriately vaccinated for age. In contrast to this 100% efficacy the efficacy of a single dose of monovalent Group A meningococcal vaccine to prevent illness in the youngest children during the 1987 epidemic period was 52% (95% confidence interval (-330%, 95%)) falling to 16% (95% confidence interval, (-538%, 90%)) after 1 year. Four cases that occurred in infants 3 to 7 weeks before the scheduled "booster" campaign supports limited true efficacy. However, the prescribed 1 to 3-month interval between the two doses in infants may be too long.(ABSTRACT TRUNCATED AT 250 WORDS)


Virulence properties of Enterobacteriaceae isolated from the small intestine of children with diarrhea. Enterobacteriaceae isolated from the duodena of Peruvian children with persistent diarrhea (PD) have been examined for virulence factors and compared with Enterobacteriaceae isolated from children with acute diarrhea, those convalescent from PD and diarrhea-free controls. Escherichia coli were isolated from 42 of 186 (23%) of the aspirates. All 11 children with PD in whom multiple E. coli colonies were examined were colonized by a single serotype. DNA probes identified enterotoxigenic E. coli in 2 of 89 (2.2%) PD aspirates and 2 of 38 (5.3%) acute diarrhea aspirates and enteroaggregative E. coli in one PD and one control aspirate. Strains positive with the enteropathogenic E. coli adherence factor probe were identified from 2 of 89 (2.2%) patients with PD and 1 of 34 (2.9%) controls. A subset of 12 E. coli strains failed to show adhesion to human duodenal enterocytes although 5 of 9 showed sparse but polar attachment to ileal cells from a child with short bowel syndrome and PD. Three of 10 Enterobacteriaceae (two E. coli, one Klebsiella species) caused diarrhea in the reversible ileal tie adult rabbit model. Colonization with virulent Enterobactericeae did not explain the majority of episodes of PD. Examination of these duodenal bacteria in the rabbit model revealed some that caused diarrhea but were not recognized pathogens.


Natural history of human immunodeficiency virus type 1 infection in children: prognostic value of laboratory tests on the bimodal progression of the disease. This study analyzes the correlation of basic laboratory test results with clinical outcome in 94 children with perinatally acquired human immunodeficiency virus 1 infection who did not receive zidovudine during the study period of 1983 to 1988. Two life-threatening conditions highly correlated with survival, opportunistic infection and severe encephalopathy, were the end points of the study. At a median age of 25 months 30 (32%) of the 94 children had developed such conditions (Group I), and their survival at 3 years of age was 48% (95% confidence interval, 24 to 72%), contrasting with the 97% survival rate (95% confidence interval, 94 to 100%) of the remaining 64 (68%) children who had not developed such conditions by age 25 months. (Group II). Compared with children in Group II, children with life-threatening complications were more likely at the onset of symptoms to be younger and have a lower CD4 count, an impaired in vitro lymphocytic proliferation and a lack of p18 or p25 antibodies in the Western blot profile and, during the progression of the disease, a negative slope of the subsequent CD4 counts. These results highlight the need for an early diagnosis of the human immunodeficiency virus 1 infection in children and demonstrate that it is possible to determine the prognosis of their disease as early as in the first year of life.


Diagnosis of human immunodeficiency virus infection by enzyme-linked immunospot assays in a prospectively followed cohort of infants of human immunodeficiency virus-seropositive women. The enzyme-linked immunospot (ELISPOT), a method for quantifying specific and total antibody-secreting cells, was used for the diagnosis of human immunodeficiency virus (HIV) infection in a prospectively followed cohort of infants born to HIV-infected women. From July 1, 1987, to June 1, 1990, 127 infants with known HIV infection status were studied. Seventeen of 22 HIV-infected infants had specific HIV-specific antibody-secreting cells (ASC). Among the infected infants rates of ASC positivity increased during the first year of life, from 25% in the first 5 days of life to 78% after 6 months. Two of the five ASC-negative infected infants were further characterized as hypo- or dysgammaglobulinemic by an adjunct ELISPOT assay for total immunoglobulin-secreting cells. Excluding hypo- or dysgammaglobulinemic infants from the analysis, the rate of ASC positivity among infected infants was 85% (17 of 20) after the age of 6 months. None of the 95 uninfected infants had a positive ELISPOT assay, including 55 who were tested in the first 3 months of life. Thus in this series the specificity was 100%. ELISPOT methodology can be a useful technique for the diagnosis of HIV infection in infants of HIV-seropositive mothers.


Lymphocyte subsets in children younger than 2 years old: normal values in a population at risk for human immunodeficiency virus infection and diagnostic and prognostic application to infected children. Data were collected prospectively from 116 children younger than 2 years old who were seen at the Duke Pediatric AIDS Clinical Trials Unit for known human immunodeficiency virus seropositivity. Forty-six (40%) of these children were human immunodeficiency virus-infected and 70 were not infected. Using 3-month blocks, 10th, 50th and 90th percentiles were calculated for the CD4+ and CD8+ cell counts, percentage of lymphocytes positive for CD4 and CD8 and T4:T8 ratios. Results from the infected and uninfected children were compared. By 3 to 6 months of age the infected patients had significantly lower CD4+ counts, percentage CD4+ cells and T4:T8 ratios, whereas the percentage of CD8+ lymphocytes was significantly higher. Absolute CD8+ counts were approximately the same in infected and uninfected children through age 2 years. Most infected children had one or more abnormal lymphocyte subset results (less than the 10th percentile for uninfected patients) by age 2: 83% had an abnormal CD4+ percentage; 78% had an abnormal T4:T8 ratio; and 67% had an abnormal CD4+ count. All 13 children who had an opportunistic infection (at any age) had an abnormal CD4+ percentage before age 2 years, and 12 of 13 had a low absolute CD4+ count or T4:T8 ratio. Among patients who died 10 of 11 had 1 or more low CD4+ count, 9 of 11 had an abnormal CD4+ percentage and 8 of 11 an abnormal T4/T8 ratio.


Ceftibuten and trimethoprim-sulfamethoxazole for treatment of Shigella and enteroinvasive Escherichia coli disease. In a prospective randomized study at two clinical sites, ceftibuten was compared with trimethoprim-sulfamethoxazole (TMP-SMX), both given orally for a period of 5 days, for the treatment of dysentery. Twenty-two children were found to have bacillary dysentery caused by Shigella and/or enteroinvasive Escherichia coli. All organisms isolated were susceptible to ceftibuten; 6 of 20 Shigella strains and 4 of 5 enteroinvasive E. coli were resistant to TMP-SMX. The diarrhea persisted for a mean (+/- SD) period of 2.4 +/- 1.4 days in the ceftibuten-treated patients vs. 3.4 +/- 1.7 days in the TMP-SMX-treated patients. The duration of fever was similar for both treatment groups. Patients treated with ceftibuten or TMP-SMX had equivalent clinical responses unless the pathogen was found to be TMP-SMX-resistant. Those who were randomized to receive TMP-SMX but who were eventually found to have TMP-SMX-resistant organisms had significantly more stools at days 3, 4 and 5 (P less than 0.02 to less than 0.00006) with more watery consistency for these days (P less than 0.02 to less than 0.005) compared to patients treated with ceftibuten. No clinical relapses were reported and no drug-related side effects were observed. We conclude that ceftibuten is at least as effective as TMP-SMX in the treatment of diarrhea caused by Shigella and enteroinvasive E. coli in children.


Colonizing and invasive strains of Streptococcus pneumoniae in Uruguayan children: type distribution and patterns of antibiotic resistance. For the first time in Uruguay the frequency of moderate to heavy colonization of the upper respiratory tract by Streptococcus pneumoniae was investigated in children with acute respiratory infections (n = 717) and in healthy controls (n = 564). Serotypes of S. pneumoniae were identified as colonizing and invasive strains and their susceptibility to antibiotics was determined. Semiquantitative cultures of nasopharyngeal aspirates yielded 42.1% of positives in ambulatory patients and 15.2% in controls. Throat swabs from hospitalized children and matched controls revealed, respectively, 18.4 and 11.5% colonization. Different sampling and culture procedures were evaluated. Seasonal variations in colonization were also detected. Geographic variations in serotype frequency and distribution were assessed. Serotype 14 was predominant among invasive and colonizing strains in patients. Ten serotypes, included in the 23-valent S. pneumoniae vaccine, were not detected. Intermediate resistance to penicillin was seldom observed, but 37.2% of the invasive strains were resistant to cotrimoxazole, indicating the need for a permanent surveillance of isolates to antibiotic susceptibility.


Epidemiology of pertussis in a developed country with low vaccination coverage: the Italian experience. In Italy pertussis vaccination is optional. Fewer than 40% of children younger than 5 years of age are vaccinated, and pertussis remains a common childhood disease. We use data from a variety of sources to examine trends and characteristics of Italian children with pertussis and reasons behind the low vaccine coverage. Approximately 25% of Italian children have experienced clinical pertussis by their fifth birthday. The disease is most severe in those less than 1 year of age; in this group an estimated 1 in 14 cases are hospitalized and 1 in 850 die. The incidence appears to be increasing in the 1- to 4-year age group despite increased vaccination coverage. The low vaccine coverage appears to be caused by the ambivalence of the Italian pediatric community about the vaccine rather than parental concerns about vaccine safety. Legislation is being considered to make pertussis vaccination mandatory.


The assembly and structure of the fibrin clot. This review summarizes our current understanding of the structure of the fibrinogen molecule, its conversion to fibrin, and the main features of fibrin clot assembly and crosslinking. Several major aspects of fibrin matrix assembly and crosslinking are newly discovered, and differ significantly from more traditional views of fibrin structure. The new information that I have summarized will be of importance for correctly understanding the role that fibrin plays in the hemostatic process, and for learning how the structure of the fibrin clot network contributes to its strength, elasticity, interactions with cells, tissue matrix, and its susceptibility to fibrinolysis.


Further evidence that heparin-dependent thrombocytopenia may result from Fc receptor-mediated interactions. Heparin-dependent thrombocytopenia (HDT) and associated thrombotic complications, are thought to be linked to the appearance of anti-platelet antibodies. Attempts were made to characterize the antibodies in the sera from 10 such patients. Western blotting against platelet antigens was inconclusive, often revealing multiple bands but with considerable variability from patient to patient. An often seen band of approximately 83 kDa was also given by some nonimmune sera and the antigen appeared to be predominantly intracellular in origin. Antibodies to major membrane glycoproteins were not readily apparent. This was confirmed by the MAIPA ("Monoclonal Antibody Immobilization of Platelet Antigens") test, performed to detect antibodies to GP Ib-IX and GP IIb-IIIa. Only one weak activity to GP Ib-IX and one weak activity to GP IIb-IIIa were detected. In contrast, an antibody to the PlA1 alloantigen was readily detected in the serum of an additional patient. The ability of HDT serum to induce the aggregation of control platelets was studied in detail. Aggregation was inhibited by EDU-3, a monoclonal antibody to GP IIb-IIIa complexes, and by the synthetic peptide RGDS, suggesting an involvement of the same pathway as used by physiologic agonists. However, in agreement with Chong et al (Thromb Res 55:291, 1989), we observed that aggregation was inhibited by rabbit IgG, suggesting that it was Fc-receptor mediated. Interestingly, 2E1, a monoclonal antibody to the Fc gamma RII receptor, induced platelet aggregation with a lag phase, a characteristic of HD antibodies. Nonetheless, for different donors, there was no correlation between the length of the lag phase induced by 2E1 and HD antibodies. Fc-receptor blockade should be considered as a means for diminishing the clinical complications of HDT.


Differential effects of extra- and intracellular calcium chelation on human platelet function and glycoprotein IIb-IIIa complex stability. The presence of extracellular ionized calcium (Ca2+) is important to support the aggregation of human blood platelets and for the stability of the platelet fibrinogen receptor, the glycoprotein (GP) IIb-IIIa complex. This study was performed in order to determine the effects of intracellular calcium chelation on human platelet functions, on the expression of the fibrinogen receptor and on the stability of the GP IIb-IIIa complex. Intracellular Ca2+ of intact human platelets was extensively chelated by incorporation of high amounts (14-50 mmol/L) of the specific Ca2+ chelator quin2 after incubation of platelets with its lipophilic acetomethoxylester form (quin2-AM). We have investigated the effects of intracellular Ca2+ chelation with quin2 on platelet aggregation and fibrinogen binding induced by ADP and human alpha-thrombin and on the stability of the GP IIb-IIIa complex studied by crossed immunoelectrophoresis of Triton X-100 platelet lysates. The results were compared with control experiments performed with intact platelets treated with the impermeant Ca2+ chelator quin2 and with EDTA or EGTA. This study shows that high intracellular concentration of quin2 inhibits human platelet aggregation and fibrinogen binding but does not induce the dissociation of the GP IIb-IIIa complex. This study adds evidence for the role of external Ca2+ to maintain the integrity of a non-dissociated GP IIb-IIIa complex and suggests that intracellular Ca2+ is not involved in the stabilization of the GP IIb-IIIa complex.


Characterization of a thrombopathy (type delta storage pool disease) affecting a pig colony. An autosomal recessive thrombopathy in pigs is described and characterized functionally, morphologically and biochemically. The affected pigs have a severe bleeding diathesis and a markedly prolonged bleeding time but normal plasma and platelet von Willebrand factor (vWF) levels. Electron micrographs and fluorescence microscopy with mepacrine reveal reduced numbers of dense granules in platelets as compared to normals. This thrombopathy is a pure delta storage pool disease (SPD), as evidenced: a) biochemically by platelet serotonin content and metabolism and by comparative ATP/ADP content and secretion; b) functionally by reduced aggregability to low concentrations of convulxin and collagen but normal aggregability to other agents and normal synthesis of thromboxane B2. The affection was first discovered in a colony of von Willebrand's disease (vWD) pigs, but is biologically and genetically distinct. It is possible to completely separate the SPD from the vWD, although originally animals could be affected by both vWD and SPD. Normal plasma and platelet alpha granule content of vWF are found in diseased animals. An intermediate disorder is also detected in animals not severely affected, which may represent the heterozygous state.


Inexpensive manual synthesis of antithrombotic peptides. We describe a simple, improved method of manual synthesis of biologically active, antithrombotic peptides. The prototype peptide KYGRGDS is an effective inhibitor of platelet aggregation in vivo. This Fmoc-based method can be implemented by the research lab that would otherwise not have access to this technology because of safety or financial concerns. This method facilitates the synthesis of large numbers of peptides in quantities of 25 to 50 mg. Fmoc manual synthesis produces high quality peptides in fewer working days than current automated approaches.


Protein C and hypertension.

Protein C and hypertension. Protein C antigen and amidolytic activity were assayed in 38 patients with essential hypertension at the first stage of disease. As compared to 20 normal healthy blood donors no abnormality was found.


Plaque fissure: the link between atherosclerosis and thrombosis. The immediate cause of arterial, predominantly coronary thrombosis is almost always cracking or fissuring of the cap of an atheromatous plaque. This exposes collagen and lipids to the flowing blood and thereby initiates thrombotic platelet aggregation, almost immediately followed by coagulation. The thrombi tend to extend into the arterial lumen, causing obstruction to blood flow and clinical symptoms and signs. Evidence for this sequence of events comes, inter alia, from angiograms of patients with unstable angina and developing myocardial infarction. Direct angioscopy in life is also visualising mural thrombi over fissured plaques in atheromatous coronary arteries. We are investigating the initial development of atheromatous plaques liable to fissuring. The cap over such plaques covers a "lipid pool". We have discovered that one factor promoting the uptake of lipid, in the form of plasma low-density lipoprotein, is the concentration of circulating catecholamines (Cardona-Sanclemente LE, Gorog P, Born GVR (1992) J Physiol London, in press). We are also investigating the immediate cause(s) of plaque fissure. We have evidence for a complex interaction of different determinants, including the concentration of macrophages, presumably as foam cells, in the plaque caps (Lendon CL, Davies MJ, Born BVR, Richardson PD (1991) Atherosclerosis 87: 87).


Neutralization of lupus anticoagulant activity by human immunoglobulin "in vitro". We studied the in vitro effect of human intravenous immunoglobulin (IVIg) on the lupus anticoagulant (LA) activity present in sera of 11 patients. LA potency was determined in all the cases and a fixed dilution of each serum was chosen to perform the dose-dependent neutralization experiments. For each patient, the dilute serum was incubated for 3 h at 37 degrees C with phosphate buffer saline (PBS) alone or containing IVIg at final concentrations of 0 to 50 mg/ml. Aliquots of the incubation mixtures were added to equal volumes of normal plasma and APTTs were performed. IVIg partially neutralized the LA activity present in 10 out of 11 patients sera. These neutralizations showed an IVIg dose-dependent behaviour. Statistically significant neutralizations were observed at least at one molar ratio (MR = [IVIg]/patient's [IgG] or [IgM]). In every case, a particular MR was found in which the neutralization was maximal (N%max). The N%max ranged from 33.6% to 79.5%. Eight patients showed maximal LA neutralization at MR ranging from 8.9 to 56.8. In one patient with drug-induced LA and another exhibiting LA cofactor effect, MRs were more elevated. We found poor negative correlation between N%max and LA potency (r = -0.46) or N%max and MR of N%max (r = 0.47), although no statistic significance was reached. However, there was good agreement between LA potency and MR of N%max (r = 0.98, p less than 0.001). We have shown that IVIg may neutralize LA activity in vitro. In view of these results, we believe that IVIg should be considered as an alternative therapy in patients with LA-related clinical complications.


Influence of aging on the activity of the hemostatic system: prothrombin fragment 1+2, thrombin-antithrombin III complexes and D-dimers in 80 healthy subjects with age ranging from 20 to 94 years. To determine the influence of aging on the activity of the hemostatic system, we measured the plasma concentration of prothrombin fragments 1+2 (F 1+2), thrombin-antithrombin III complexes (TAT) and fibrin degradation fragments D-dimers (D-D) in 80 healthy subjects with age ranging between 20 and 94 years. All subjects were free of acute or chronic diseases. The three markers (semi-log scale) were positively correlated with age (r greater than 0.7, p less than 0.01). Mean plasma levels of F 1+2, TAT and D-D were two- to five-fold higher in subjects with age greater than or equal to 60 as compared to those less than 60 years (n = 40 in both groups). In these two groups, normal values for F 1+2, TAT and D-D ranged from 0.3-1.2 vs 0.7-2.4 nmol/l, 1.4-2.6 vs 1.9-6.4 micrograms/l and 33-433 vs 312-1180 micrograms/l, respectively. Sex did not influence the results. We conclude that the activity of the hemostatic system is markedly age-dependent, and that elderly subjects display a biological picture of "prethrombotic" state. In addition, for a right clinical use of these three markers, age should be taken into account when normal range is to be established.


Impaired fibrinolytic response to DDAVP in patients with von Willebrand's disease. The fibrinolytic response after infusion of 1-deamino-8-d-arginine vasopressin (DDAVP) was examined in 4 patients with the severe form and 17 patients with the moderate form of Von Willebrand's disease (VWd) and compared to that of 9 normal subjects. Tissue plasminogen activator (t-PA) antigen (Ag) and activity and tissue plasminogen activator inhibitor (PAI) were measured before and after DDAVP. t-PA Ag was found to be significantly higher before DDAVP in patients than in controls. No release of t-PA Ag was observed in the 4 patients with the severe form of VWd but increased release was observed in patients with moderate forms. However, t-PA released into the circulation in these 17 patients had a lower functional activity as compared to that of normal subjects. PAI was significantly lower in patients before DDAVP than in normal subjects and the decrease in PAI after DDAVP was significantly less in patients than in controls. It is concluded from this study that patients with VWd have an abnormal fibrinolytic response after stimulation regardless of the severity of the disease. Furthermore, the results suggest either that patients with VWd have a double defect in VWF and tissue plasminogen activator or that the primary deficiency of VWF influences the synthesis and/or release of t-PA by endothelial cells.


Polymorphism of platelet glycoprotein Ib associated with variability of the 85-kDa macroglycopeptide region. Polymorphism of platelet membrane glycoprotein Ib (GP Ib) has been described in both normal and functionally abnormal platelets. In this report we have investigated the polymorphism in one Australian Caucasian family with normal platelet function in which the following phenotypes were found: father, BC; mother, CD; son, BC; and daughter, BD; thus establishing a genetic basis for this phenomenon. The daughter's phenotype BD is highly distinctive due to the double-band pattern obtained by gel analysis. Platelets from the daughter were digested with either trypsin or elastase and the GP Ib cleavage products were examined by immunoprecipitation with anti-GP Ib-IX complex monoclonal antibodies against each of the major tryptic domains. On the basis of these studies, we have determined that the GPIb polymorphism in this family resides in the 85-kDa, macroglycopeptide region of the alpha-chain of GP Ib and not in either the 45-kDa, N-terminal region of GP Ib alpha or the membrane-associated region of the complex.


A directed search for mutations in hemophilia A using restriction enzyme analysis and denaturing gradient gel electrophoresis. A study of seven exons in the factor VIII gene of 170 cases. Genomic DNA from 170 unrelated hemophilia A patients was examined for gene defects in the coding region of the Factor VIII gene. Exons 18, 22-24 and 26 contain a CGA codon for arginine within the recognition sequence for the restriction enzyme Taq I. These five sites were amplified by the polymerase chain reaction and tested for abnormal Taq I restriction patterns. In five cases, the enzyme Taq I failed to digest the amplified fragments. Direct sequencing of the amplified products demonstrated a C to T transition in the coding strand of exons 18, 22 and 24 in three severe hemophilia A patients resulting in TGA termination codons. Two patients showed G to A transition in exons 24 and 26 reflecting a C to T transition in the non-coding strand substituting a glutamine for an arginine. Three deletions involving exon 26 and one exons 23-26 were found in severe hemophiliac patients. In contrast, exons 23 and 24 failed to amplify in one patient with a moderate form of the disease suggesting an in-frame splicing of exons 22 and 25. Exon 8 and the 3' end of exon 14 were analyzed by denaturing gradient gel electrophoresis (DGGE). Two patients with a moderate form of the disease demonstrated an abnormal electrophoretic pattern in exon 8 and sequencing demonstrated missense mutations at codon 372 for arginine within a thrombin activation site. One missense mutation was a C to T transition substituting cysteine for arginine and the other was an infrequent G to C transversion at an adjacent nucleotide changing the same arginine to proline.(ABSTRACT TRUNCATED AT 250 WORDS)


Immunological study of platelet glycoproteins and alloantigens in families of patients with Glanzmann's thrombasthenia. To explore the genetic transmission of Glanzmann's thrombasthenia (GT) we conducted a study of platelet glycoproteins and antigens using monoclonal and polyclonal antibodies and cytofluorometry. Twenty-one members of 4 different families in which at least one member was affected with GT were explored. The reactivity with AP2 (anti-GP IIb-IIIa complex, CD41a), AP3 (anti-GP IIIa) and antibodies against HPA-1a (PLA1) and HPA-3a (Leka) was lower than 5% in the platelets of 5 patients with GT type I. Nine obligatory carriers of the GT trait were tested by the same technique. The mean values of reactivity expressed as per cent of normal with AP2 (anti-CD41a) and AP3 (anti-GPIIIa) were 58.3 +/- 3.6% and 58.6 +/- 3.6% respectively, while the frequencies of HPA-1a (PLA1) and HPA-3a (Leka) negativity were 0/9 and 4/9 respectively. When both antigens were expressed they were detected at reduced levels. These results indicate that cytofluorometry is a simple technique for GT detection, but this method would nevertheless not appear to be sufficiently accurate to identify obligatory carriers in a general population.


Platelet plasma membrane is equally distributed between surface and osmotically-evaginable surface-connecting membrane, independent of size, subpopulation or species. Osmotic swelling can double the external plasma membrane surface area of normal human platelets. The surface-connected open canalicular system (SCCS) has been proposed as the major source for this additional membrane. As bovine (B) platelets have been reported to lack SCCS, we compared osmotic swelling for B and human (H) cells. Addition of water to platelet-rich-plasma (10-90% v/v) caused sequential shape change and osmotic spherocyte (OS) formation, analyzed for size and surface area changes from time-dependent phase-contrast videomicroscopic images. Selected samples were fixed and stained with tannic acid prior to osmic acid fixation for visualization of open SCCS by transmission electron microscopy. Bovine platelets required 3-4x less water dilution of PRP than human platelets, with significant OS forming at 20% water addition. Continued water dilution converted 50% of platelets to OS, with maximally stable swelling and no significant lysis for bovine OS up to 60% dilution. Electron micrographs of unactivated discocytes (D) and of optimally-swollen OS showed open SCCS in human D not detectable in any of the swollen platelets, though granules, mitochondria and a small number of vesicles and vacuoles persisted. No evidence for any open SCCS was found for bovine D or OS, though the OS otherwise appeared similar to human OS. Geometric measurements of D and nonlysed OS showed a stable, maximal 2.1 +/- 0.1 fold increase in external plasma membrane surface area with osmotic swelling, identical for different-sized normal human platelets (mean volume V = 2.8-6.8 fl), for abnormally-large platelets (V = 8.7-11.6 fl), or for bovine or rabbit platelets (V = 3-4 fl). As osmotic swelling appears to primarily externalize SCCS in human platelets, the identical relative amounts of internal membrane externalized for bovine platelets is hypothesized to arise from an osmotically more labile, "closed", and structurally simpler SCCS or from a distinct membrane source than in human platelets. It appears that the surface to invaginated ("SCCS") plasma membrane is kept constant for human, bovine and rabbit platelets, independently of platelet production, size-dependent subpopulations, or of platelet ageing.


A path model of type A and type B responses to coping and stress in employed black women. The purpose of this study was to test a conceptual model of the effects of age, weight, type, cholesterol, systolic blood pressure, and coping on the stress response of southern black women. Data were collected on 280 employed black women. Audiotape analyses of the structured interview resulted in 136 Type A's and 144 Type B's. Each individual was screened for hypercholesteremia using a portable reflectance photometer. Blood pressure measurements were taken every minute before, during, and after a stress-producing Type A/B interview. Type A's had higher resting systolic blood pressures, higher cholesterol levels, higher levels of stress during the interview, and lower coping scores than Type B's. The relationships between age, weight, and resting systolic blood pressures all were in the direction reported in the literature.


Model for predicting work performance among persons with multiple sclerosis. A causal model was tested on a sample of 201 MS subjects for the purpose of examining the role of selected demographic factors, work impediments, and work enhancers as predictors of activities essential for performing outside employment, home making, and for personal care. The results showed a relatively good fit of the model to the data through a number of indicators. Explained variance included: 9.0% in work impediments from the direct effect of age; 32.3% in work enhancers--A (job and environmental adjustment, social support, healthful practices) through direct effects from MS duration and work impediments; 9.8% in work enhancers--B (personal attributes) through direct effects from MS duration and work impediments; 61.1% in work performance through direct effects from the antecedent variables (age, education), stress-appraisal variable (work impediments), and coping variables (work enhancers-A, work enhancers-B).


Increasing condom-use intentions among sexually active black adolescent women. Whether a social cognitive theory AIDS prevention intervention would increase intentions to use condoms among 109 sexually active inner-city black female adolescents was tested. Analyses revealed that the women scored higher in intentions to use condoms, AIDS knowledge, outcome expectancies regarding condom use, and self-efficacy to use condoms after the intervention than before the intervention. Although increased self-efficacy and more favorable outcome expectancies regarding the effects of condoms on sexual enjoyment and sexual partner's support for condom use were significantly related to increased condom-use intentions, increases in general AIDS knowledge and specific prevention-related beliefs were not.


Physiological and psychological variables related to functional status in chronic obstructive pulmonary disease. The relationship of selected physiological and psychological variables to functional status in patients with chronic obstructive pulmonary disease (COPD) was investigated in this study. There has been limited exploration of the relative contribution of these variables to the performance of activities of daily living in this population. A convenience sample of 104 outpatients with COPD participated in the study. The independent variables were causal attributions, depressed mood, self-esteem, pulmonary function, and exercise capacity, and the dependent variable was functional status. There was a significant difference in functional status between those who did and those who did not ask the question, "Why me?" (p = .03). In addition, the remaining psychological variables and both physiological variables were significantly correlated with functional status (p less than or equal to .01). To identify the combination of study variables most predictive of functional status a multiple regression analysis was performed. The combined variables of exercise capacity and depression best predicted functional status (p less than .0001). It is concluded that both physiological and psychological factors are important in understanding functional status in this population.


Standardization of a device to measure unsupported arm exercise endurance in chronic obstructive pulmonary disease. The purpose of this study was to compare respiratory responses with moderate and slow rates of unsupported arm exercise (UAE) with a newly developed electromechanical device. Twenty-one patients with chronic obstructive pulmonary disease (COPD) were studied. Exercise endurance limits, metabolic, ventilatory and sensation outcomes were determined at rest prior to exercise and at end-exercise endurance limits. Increases from baseline rest for both exercise rates were observed in: oxygen uptake, carbon dioxide production, inspiratory flow, minute ventilation, respiratory rate, dyspnea, respiratory effort, and arm fatigue. Endurance limits were similar for both rates of UAE. These data provide standards against which UAE in COPD can be evaluated.


Psychiatric rehospitalization of the severely mentally ill: patient and staff perspectives. Research findings about psychiatric rehospitalization of severely mentally ill persons have provided few insights into its meaning for patients and staff. An attributional analysis of the causes schizophrenic patients and their staff gave for rehospitalization was used to test the limits of attributions theory in predicting how the event would be viewed. Results indicated that patients tended to give internal attributions for their readmission and believed that the cause was not under their control. Staff also attributed the problem to something internal to the patient, however, they were likely to see the cause as patient effort, which is under patient control.


Temperature and temperature measurement after induced hypothermia. This study was designed to assess factors associated with afterdrop, the fall in core temperature following completion of cardiac surgery, and determine the validity of noninvasive measures of temperature to predict core temperature in the severely hypothermic patient. Twenty-five postcardiac surgery patients served as subjects. Core temperature was measured using the pulmonary artery, bladder, and esophageal sites. The less invasive measures included a tympanic membrane thermometer, oral and axillary electronic thermometers, and a forehead surface temperature indicator. Temperatures were recorded every 10 minutes for 2 hours. End-of-surgery temperatures ranged from 30.3-38.3 degrees C (86.5-100.9 degrees F) with a mean of 36.02 degrees C (96.84 degrees F). Temperature change over the next hour ranged from a rise of 2.5 degrees C (4.5 degrees F) to a fall of 4.1 degrees C (7.2 degrees F). Factors associated with afterdrop included age, end-of-surgery temperature (both positively) and body mass (negatively). No noninvasive measure appeared to be a valid indicator of core temperature in these hypothermic patients.


[Markers of HBV infection in drug addicts]. Prevalence of markers of HBV infection was tested in two cohorts of drug addicts: in 140 addicts whose sera were drawn in the years 1985-1986 and in 100 addicts whose sera were drawn in the years 1988-1989. HBsAg was found in 10 (7%) patients in the earlier cohort and in 12 (12%) addicts in the latter, while at least one marker of HBV infection was present in 39% and 69%, respectively No correlation was found between the presence of markers of HBV infection and age, gender, or HIV status, however, duration of drug abuse increased the risk of HBV infection. There was no difference in mean titres of anti-CMV and anti-HSV-1 between subjects with and without HBV markers what suggests that the tested markers were specific.


Factors influencing psychiatric nurses' use of section 5(4). A review of the literature suggests that little is known about the factors influencing the use of nurses' holding power under Section 5(4) of the 1983 Mental Health Act. This study aimed to redress this gap. Data were collected over one year by analysing the statistical returns of all wards within one mental health unit that used Section 5(4). It was found that it was used in the management of 3.8% of admissions. All but 4% of those detained were converted to an extended section. A number of variables were explored, the results of which suggest that psychiatric nurses' decision to invoke this section of the Act is not simply a case of patients being a danger to themselves or others but rather reflects an interplay of a number of factors. An awareness of such factors may reduce the necessity to implement this emergency nursing intervention.


[Effects of alpha and beta receptor blockaders on urinary excretion of sodium and potassium in healthy rats and in those with chronic renal failure]. The aim the study was to investigate the effects of both alpha and beta adrenergic blockade on sodium and potassium excretion in rats with chronic renal failure. The chronic renal failure was induced by surgical ablation of 75% renal mass. Controls were shame operated rats. All experiments were performed under sodium pentobarbitone anesthesia. Blood pressure was measured intraarterially and glomerular filtration rate was estimated as inulin clearance. Phentolamine (4 micrograms/kg/min)--Group I; or propranolol (4 micrograms/kg/min)--Group II were infused i.v. to controls and chronic failure rats. alpha-receptor blockade with phentolamine lowered GFR and blood pressure to the same extent in controls and rats with chronic renal failure. alpha-blockade caused increased fractional sodium excretion only in controls while urinary excretion of sodium and potassium fell in rats with chronic renal failure. The beta-blockade with propranolol (4 micrograms/kg/min)--Group II did not affect both blood pressure and GFR either in controls nor in rats with chronic renal failure. However propranolol infusion increased sodium and potassium excretion in controls but not in rats with chronic renal failure. Our study showed the divergence in the role of adrenergic receptors in regulation of urinary electrolytes excretion between controls and rats with chronic renal failure.


[Effects of alternating as well as long-term intensive polychemotherapy in the remission-maintaining treatment of patients with acute myelocytic leukemia]. The paper presents the results of 2 year maintenance treatment of patients with acute myeloid leukemias using cyclic, rotating and intensive polychemotherapy. Complete remission (CR) was achieved in 22 out of 33 patients with doxorubicin (60 mg/m2; 1-3 days) and cytosine arabinoside (100 mg/m2; 1-7 days). All patients in CR entered into the authors' programme of intensification therapy consisting of two consecutive polychemotherapy cycles, using various combinations of amsacrine, doxorubicin, cytosine arabinoside, etoposide and 6-thioguanine repeated every 3-4 months. The mean survival so far is 18 months (range, 1-52 months). The projected 3-years survival rate in complete remission concerns 33% of the patients treated. Relapses occurred in 50% of the patients, within 10 months (range 1-35 mos.) after CR. Our results indicate that intensive cyclic polychemotherapy markedly prolongs the survival of AML patients. The high frequency of relapses during the first year after CR indicates the necessity to enhance the degree of aggressiveness of treatment shortly after a complete remission has been obtained.


[Recurrent benign intrahepatic cholestasis syndrome]. Up to now about 100 cases of benign recurrent intrahepatic cholestasis have been reported. In this case we give a description of familial form of this disease, in which cholestasis was associated with inflammatory liver response. A brief review of this rare condition and differential diagnosis have been presented.


[Effect of salbutamol on heart rate in patients with atrial fibrillation]. The influence of salbutamol aerosol on heart rate in patients with chronic atrial fibrillation was examined. The study comprised 15 patients (9 women and 6 man) in age 47-79 (mean 60) treated because of chronic obstructive pulmonary disease (COPD) with concomitant atrial fibrillation. The influence of salbutamol on heart rate was investigated during six hours ecg Holter monitoring (3 hours before and 3 hours after using salbutamol). The analysis showed that salbutamol aerosol in two doses increased heart rate calculated as mean heart rate per hour from 3-hours intervals from 4061 congruent to 1022.5 to 4185 congruent to 976 (p greater than 0.05). Mean value of blood pressure in one hour intervals did not change significantly. In patients with concomitant chronic atrial fibrillation and COPD salbutamol increased heart rate but difference between mean values before and after administration was not significant.


[Clinical value of indirect automatic blood pressure determination in patients with atrial fibrillation]. The value of indirect blood pressure in patients with atrial fibrillation was studied. Three methods were compared; single measurement, mean of three measurements and mean value of automatic measurement during 2-hours and whole day. The study comprised 13 patients; 7 men and 6 women aged 52-77 (mean 62.8 congruent to 6.6) hospitalized for cardiac diseases. The measurement were done using the mercury sphygmomanometer and auscultatory Korotkoff technique and automatic blood pressure monitoring (Acutracker--Medilog Oxford). The investigations showed that systolic and diastolic blood pressure during 24-hours monitoring were lower than single or repeated three measurements. The highest correlation coefficient to 24-hours recording was found for the mean value of 2-hours recording. It seems that blood pressure measurement in patients with atrial fibrillation (3-times in few minutes intervals) should be verified by using prolonged automatic recording.


[Comparison of the effects of sodium fluoride and calcidiol on the remodeling of the endosteal surface of the rib cortex in dogs after administration of prednisone]. The effects were compared of sodium fluoride and calcidiol on the remodeling of the rib cortical-endosteal surface of dogs treated with prednisone for long time periods. The study used a histomorphometric and tetracycline labeling methods. It was found that administration of sodium fluoride with calcidiol and calcium carbonate limited in a higher degree than the treatment with calcidiol and calcium carbonate the development of the osteoporotic changes induced by glucocorticoids. This included reduced enhancement of the bone resorption surface, increased bone formation surface and osteoid thickness connected with acceleration of mineralization rate. The changes induced by sodium fluoride had a favorable effect on the ratio of the resorption to the formation process at the sites of bone remodeling.


[Manometric evaluation of the effects of intravenous administration of glucagon, buscopan and papaverine on the contractile activity of the sphincter of Oddi]. Endoscopic manometric technique was used to investigate the effects of spasmolytic drugs on the sphincter of Oddi (s.O.) motility. 41 patients were randomly divided into 4 groups. In every patient the characteristics of the s.O. was monitored before and during 5 min. period after i.v. administration of: 20 mg buscopan, 1 mg glucagon, 40 mg papaverine or 2 ml of 0.9% NaCl solution. After buscopan administration the amplitude and frequency of phasic contractions of the s.O. were decreased as well as a baseline pressure in the s.O. Glucagon reduced frequency and amplitude of phasic contractions of the s.O. without influencing the baseline pressure. Papaverine reduced only frequency of phasic contractions. Physiological saline caused no change in pressure characteristics of the s.O.


[Effect of bezafibrate on serum lipid, lipoprotein and apoprotein levels in patients with hyperlipoproteinemia type II and IV]. The effect of six months' treatment with bezafibrate (400-600 mg daily) on serum lipids, lipoproteins and apolipoproteins concentrations was investigated in 32 patients with primary hyperlipoproteinemia (HLP) type IIa, IIb and IV. In all types of HLP the reduction of serum cholesterol (CH), triglycerides (TG), very low density lipoprotein (VLDL) fraction and increase in high density lipoprotein (HDL) cholesterol, HDL3-CH and apolipoprotein AI concentrations was observed. In type IIa and IIb HLP low density lipoprotein (LDL) cholesterol and apolipoprotein B levels decreased while in type IV increased. The greatest fall of serum CH concentrations was observed in type IIb while of serum TG level--in type IV. In 15 (46.9%) patients the side effects were noted. They were: abdominal pain, skin rash, myalgia, moderate increase in CPK and aminotransferase activities. Most of there side effects disappeared spontaneously without treatment cessation.


[Bactericidal activity of plasma and leukocytes in patients after kidney transplantation]. In renal transplant patients the plasma and leukocytes bactericidal activity was estimated towards the vivid bacterial cells of the standard strain of Staphylococcus aureus Oxford 209P. Considering the kind of the immunosuppression used, the patients were divided into two subgroups. The first subgroup comprised the patients treated with cyclosporin A and prednisone (12 persons) and the second one--the patients treated with azathioprine and with prednisone (19 persons). The control group consisted of 20 healthy persons. In the renal transplant patients significantly more bacterial cells survived the incubation with plasma and peripheral blood leukocytes and the reduction of bactericidal activity of the leukocytes alone was also found. The reduction of bactericidal activity of the peripheral blood leukocytes observed in the patients of the group tested may have resulted from the inefficient microbicidal systems of those cells. The kind of the immunosuppressive treatment did not seem to affect the bactericidal activity of the leukocytes and plasma examined in the renal transplant patients peripheral blood. It may be supposed that the increased susceptibility of those patients to bacterial, viral and mycotic infections is due in part to the reduced bactericidal capability of the peripheral blood leukocytes.


[Successful nitrogranulogen treatment of severe nephrotic syndrome in glomerulonephritis--case report]. A patient with mesangio-proliferative glomerulopathy with advanced nephrotic syndrome (proteinuria 20 g/day) received bolus doses of corticosteroids (5 x 1.0 g iv) but renal function continued to deteriorate. A 11-day course of nitrogranulogen (0.01 mg/kg/day) with prednisone (0.5 mg/kg) caused complete disappearance of proteinuria with improvement in renal function. This observation supports our earlier findings indicating that treatment with low dose nitrogranulogen may be efficacious in some forms of glomerulopathies.


[Drug consumption by the patients with ischemic heart disease (IHD) studied by the Pol-Monica Warsaw and the effect of selected factors on the treatment of IHD]. As part of a multicentre WHO MONICA-Project the present drug consumption by Warsaw population was assessed (evaluated by DDD). In Warsaw population 2646 inhabitants aged 35-64 years were examined. Ischaemic heart disease was diagnosed in 30% screened persons. Only 39% of subjects were involved in taking cardiovascular drugs. In ischaemic heart disease the drugs used most frequently were pentaerythritol tetranitrate and dipyridamole. The influence of several psychosocial factors on the fact of undertaking of pharmacological therapy was analysed using multivariant logistic function method. Significant relationships were found for age, health self-care self-estimation of health status and fact of medical consultation. Persons with high values of MLF (fifth quintile) underwent drug treatment six time more often, then persons with low values of MLF (first quintile).


[Blood lipids in patients after kidney transplantation]. In 50 kidney transplant patients [KTP] [37 of them were treated by cyclosporine A and prednisone--CyA group while 13 of them by azathioprine + prednisone--Aza group] serum levels of total cholesterol, LDL-cholesterol, HDL--cholesterol and triglyceride et different time intervals during the first three years after transplantation. The control group comprised 20 healthy subjects. KTP were characterized by significantly elevated plasma levels of total cholesterol, LDL-cholesterol and triglycerides. In KTP treated by CyA+prednisone even an increase of plasma total cholesterol and LDL-cholesterol and a transitory decline of HDL--cholesterol were found as compared with respective basal values. From results obtain in this study it follows, that kidney transplantation does not normalize the lipid profile in KTP, and that KTP treated by CyA are characterized by more marked lipid abnormalities than those on azathioprine therapy. Time after successful kidney transplantation does influence significantly the lipid index of KTP.


[24-hour ECG monitoring by the Holter system in early diagnosis of coronary disease]. The aim of this study was to investigate the value of continuous ECG monitoring in early diagnosis of coronary heart disease. 312 patients (59 women, 253 men, aged 20-60 years) with chest pain since 3-6 months were studied. All patients underwent clinical examination, serum lipid determination, exercise ECG test, ambulatory ECG monitoring and coronary angiography. 194 patients had normal or nonsignificant irregularities of coronary arteries, in 118 patients significant atherosclerotic stenoses were found. Sensitivity and specificity of ECG monitoring were: 46 and 54% respectively. Significantly higher frequency of changes and longer duration of depression or elevation of ST segment in patients with significant coronary artery stenosis was found. ST segment changes without angina were observed in 23,7% of patients.


[Exertion tolerance in the early period after myocardial infarction, the results of echocardiographic examination and the clinical course of infarction]. The relation between exercise test, echocardiography and clinical course of acute myocardial infarction was investigated. 17-34 days after an acute myocardial infarction, before hospital discharge, 58 patients underwent exercise test and 17-28 days- echocardiography. Low exercise capacity was significant related to angina before infarction, maximal CKNAC and complications during clinical course. There was inverted correlation of asynergy index calculated from echocardiography with maximal workload achieved during exercise test.


[Effect of metabolically uncompensated diabetes mellitus on mononuclear cell populations in peripheral blood]. The study has been aimed at comparison of the number of peripheral blood mononuclear white cells in the E. SRBC, EA, EAC, SIg markers in patients with insulin dependent (type 1) and non-insulin dependent (type 2) diabetes and healthy donors. There was no marked difference in the numbers of surface markers E SRBC and SIg among the 3 groups. Significant increase in the number of cells with markers: FcIg and C3 fragment of complement was noted in patients with diabetes type 1 and type 2. No relation between the number of mononuclear cells in peripheral blood and the blood glucose level was observed.


[Does hormonal activity of pheochromocytoma affect long-term prognosis in surgically treated patients?]. 89 patients were operated upon for pheochromocytoma. 61 patients (37 women and 24 men) were available for follow-up. Mean age at operation was 39 +/- 12.3 yrs. Before operation paroxysmal and sustained hypertension were observed in 34 (55.7%) and 27 (44.3%) patients, respectively. Postoperatively permanent normalization of blood pressure was achieved in 38 cases (62.3%). All patients were divided into four groups. Group I with increased urine excretion of noradrenaline (NA) and adrenaline (A) consisted of 19 (31.1%) patients. Group II (increased NA excretion) included 27 (44.3%) patients. Group III comprised 3 (4.9%) cases with elevated A excretion. 12 (19.7%) patients with normal urinary excretion of catecholamines and increased excretion of methoxycatecholamines were alloted to group IV. Permanent normalization of blood pressure predominantly was observed in group IV--91.6%, whilst in group I, II and III this hypotensive effect was noted in 57.9%, 51.8% and 2/3, respectively. Permanent normalization of blood pressure can be mainly expected in patients with pheochromocytoma in whom catecholamines are rapidly inactivated within the tumor.


A vasotocin-like peptide in Aplysia kurodai ganglia: HPLC and RIA evidence for its identity with Lys-conopressin G. The presence of a vasopressin (VP)- or vasotocin (VT)-like peptide in the central nervous system of the gastropod mollusc Aplysia has been indicated previously. In the case of Aplysia californica, HPLC and RIA evidence suggested the peptide was VT-like but not identical with the nonmammalian vertebrate peptide [Arg8]VT (AVT). In the present study, anterior ganglia extracts from the related species Aplysia kurodai were analyzed by HPLC followed by RIA. Further analysis of the major AVT-IR peak showed it to be indistinguishable, in three distinct solvent systems, from the sea snail venom peptide Lys-conopressin G, but to be different from the vertebrate peptides [Arg8]VP (AVP), [Lys8]VP (LVP), AVT, oxytocin (OT), mesotocin, isotocin, aspargtocin, glumitocin, and valitocin, from the sea snail venom peptide Arg-conopressin S, and from the peptides [Lys8]VT and [Gln8]OT. In addition, the carboxymethylated (CM) A. kurodai peptide had the same HPLC retention time as CM-Lys-conopressin G. The HPLC/RIA results suggest that (i) based on the properties of the solvent systems used, the A. kurodai peptide has two basic amino acids (like the conopressins but unlike the vertebrate peptides), and (ii) there is a high probability that the A. kurodai peptide is identical with Lys-conopressin G.


Effect of a single dose of des-glycinamide-[Arg8]vasopressin or oxytocin on cognitive processes in young healthy subjects. A single dose of des-glycinamide-[Arg8]vasopressin (DGAVP, 2 mg intranasal) or oxytocin (OXT, 20 IU intranasal) was given to female and male volunteers, respectively, in a placebo-controlled double-blind trial. Memory, vigilance, attention, and mood were tested starting 10 minutes after treatment. The DGAVP dose improved delayed recognition of abstract words when measured 1 week after treatment and reduced the intercept of a memory comparison task (Sternberg paradigm). A trend was present for DGAVP and OXT to affect learning, i.e., storage processes of verbal memory in an opposite way; DGAVP improved, while OXT attenuated initial storage and the rate of storage. No treatment effects on visual memory and vigilance were found. Of the mood measures, vigor was reduced immediately after treatment with OXT.


Isolation of FMRFa-like peptides from the DB-containing connective tissue of Helix aspersa. The endocrine dorsal bodies (DB) of Helix aspersa are innervated by axons from the central nervous system, which establish synapse-like structures (SLS) with the DB cells. Previous immunocytochemical studies suggested the presence of FMRFa-like substances in nerves of the DB area and in SLS. This paper reports on biochemical attempts undertaken in order to investigate the nature of these substances: the use of HPLC and RIA confirms the presence of three FMRFa-like peptides in the DB-containing connective tissue among which one is probably the FMRFa itself.


Linear and cyclic beta-casomorphin analogues with high analgesic activity. We investigated the antinociceptive efficacy of casomorphin (CM) derivatives using the vocalization test. Male Wistar rats received chronic microcannulae into the right lateral ventricle. One week later we examined the analgesic effect of CM derivatives 10, 30, 60, and 90 min after intraventricular injection (5 microliters). The analgesic effect was calculated as the individual percent increase in the pain threshold and was compared to controls (saline treatment). The substitution of D-lysine and D-ornithine in position 2 in connection with a cyclization through ring closure of the 2 position side chain amino group to the C-terminal glycine-COOH group resulted in high analgesic potency. The substitution of D-Pro4 was without any effect in the ineffective linear derivatives and decreased the effectiveness in the highly effective cyclic derivatives. The cyclic [D-Orn2]CM-5 and the cyclic [D-Lys2]CM-5 are the CM derivatives with the highest antinociceptive activity. The cyclic [D-Orn2]CM-5 is greater than 1000 times more effective than morphine. We conclude, on the basis of studies of receptor binding and in vitro investigations, that mu receptor activity alone is not responsible for the analgesic activity. The delta receptor and possibly also the kappa receptor could modulate the nociceptive effectiveness.


Structural characterization of neuropeptide Y from the brain of the dogfish, Scyliorhinus canicula. A peptide of the pancreatic polypeptide (PP) family was isolated in pure form from the brain of an elasmobranch fish, Scyliorhinus canicula (European common dogfish). The primary structure of the peptide was established as: Tyr-Pro-Ser-Lys-Pro-Asp-Asn-Pro-Gly-Glu10-Gly-Ala-Pro-Ala-Glu-Asp- Leu-Ala-Lys- Tyr20-Tyr-Ser-Ala-Leu-Arg-His-Tyr-Ile-Asn-Leu30-Ile-Thr-Arg- Gln-Arg-Tyr-NH2. This sequence contains only two amino acid substitutions compared with pig neuropeptide Y (NPY) (Gly for Asp11 and Lys for Arg19), and two substitutions (Gly for Asp11 and Leu for Met17) compared with frog NPY. The amino acid sequence of NPY from dogfish brain is appreciably different from the neuropeptide Y-related peptide previously isolated from dogfish pancreas (five amino acid substitutions). The data indicate that evolutionary pressure to conserve the complete primary structure of neuropeptide Y has been very strong. It is suggested that the NPY-related peptide present in the pancreas of elasmobranch and teleost fish represents the piscine equivalent of mammalian peptide tyrosine tyrosine (PYY).


Biological effects of human gastrin I and II chemically modified at the C-terminal tetrapeptide amide. Binding to gastrin receptors and gastric acid secretion experiments were performed with gastrin derivatives modified at the C-terminal tetrapeptide amide from HG-13 sequence. 1. When the ultimate phenylalanine amide was replaced by a phenethylester or a phenetylamide moiety, the resulting compound bound to gastrin receptors (Kd approximately 10 nM) and exhibited antagonist activity on gastrin-induced acid secretion in the anesthetized rat. 2. Changing the peptide bond between Trp and Leu residues to a -omega(CH2-NH)- bond resulted in a compound which also bound to gastrin receptors (Kd approximately 10 nM) but presented agonist activity on acid secretion in the rat. In contrast, when the peptide bond between Leu and Asp residues was replaced by a -omega(CH2-NH)- bond, the resulting compound was devoid of any affinity for gastrin receptor (Kd greater than 10(-6) M) and of any biological activity. 3. The HG-13 derivatives were synthesized in sulfated and unsulfated forms: O-sulfation of the HG-13 tyrosine residue did not change its intrinsic in vivo activity but enhanced its affinity for gastrin receptors (Kd approximately 0.3 nM). On the contrary, O-sulfation of the various chemically modified HG-13 had no significant effect in either in vitro or in vivo experiments. 4. Finally, no significant difference between binding on parietal (F3) and nonparietal (F1) cells was observed, in agreement with the presence of a gastrin-type receptor in these two cell populations.


Balance of glutamate and dopamine in the nucleus accumbens modulates self-stimulation behavior after injection of cholecystokinin and neurotensin in the rat brain. Subpopulations of dopamine (DA) neurons in the ventral mesencephalon have been reported to contain cholecystokinin (CCK) and neurotensin (NT), giving rise to DA, DA/NT, NT/CCK and DA/CCK/NT projections. More precisely, colocalized DA/CCK neurons project mainly to the caudal part of the medial nucleus accumbens, whereas its rostral portion receives CCK and DA nerve terminal networks that are structurally independent. We investigated the respective effects of both CCK and NT on the intracranial self-stimulation behavior (ICSS) from the posterolateral hypothalamus after their direct administration into the lateral ventricle (ICV), into both portions of the nucleus accumbens, into the ventral tegmental area (VTA), and into the subiculum of the hippocampal formation (SUB). The ICV injection of 150 pmol CCK8 induced a decrease in the rate of ICSS. By contrast, the direct administration of 150 pmol CCK8 into the mediocaudal part of the nucleus accumbens induced an enhanced rate of ICSS while a similar injection into its rostral portion gave rise to a slight transient decrease of ICSS. When injected into the SUB, both CCK8 and glutamate produced decreased rates of ICSS at femtomolar doses one thousand-fold under the picomolar concentrations used for ICV injections. Neurotensin induced similar behavioral profiles to that observed after the ICV injection of CCK8 or into both portions of the nucleus accumbens. Neurotensin and CCK8 displayed opposite effects on ICSS when administered into the SUB or into the VTA, suggesting they may regulate ICSS most probably through different synaptic mechanisms and through different anatomical pathways.


Hypothalamic neuropeptide Y disturbances in the obese (cp/cp) JCR:LA corpulent rat. Regional hypothalamic neuropeptide Y (NPY) concentrations were compared between cp/cp JCR:LA corpulent rats, which were grossly obese, hyperphagic, and hyperinsulinemic, and lean (+/+) controls. In freely fed cp/cp rats, NPY levels in the arcuate nucleus (ARC) were 31% higher than in lean rats (p less than 0.001). In lean rats, chronic food restriction significantly raised NPY levels by 22% in the ARC (p less than 0.05) and by 44% in the dorsomedial nucleus (DMH; p less than 0.05). By contrast, food-restricted cp/cp rats showed no change in the ARC, but NPY levels rose in the DMH (by 36%; p less than 0.05) and ventromedial nucleus (31%; p less than 0.05). Increased NPY levels in the ARC, the major site of hypothalamic NPY synthesis, suggests increased NPYergic activity in cp/cp rats; given the central actions of NPY, this could contribute to hyperphagia, obesity, and hyperinsulinemia in this syndrome. Abnormal NPY responses to food deprivation further suggest dysregulation of NPY in cp/cp rats.


Uptake and metabolism of [3H]-Leu-enkephalin following either its intraperitoneal or subcutaneous administration to mice. The uptake and metabolism of 30 micrograms/kg [3H]-Leu-enkephalin ([3H]-LE) following either intraperitoneal (IP) or subcutaneous (SC) administration to Swiss Webster mice was examined. Uptake of [3H] was rapid, with peak levels of radioactivity in plasma observed at 5 or 10 min following IP or SC peptide injection, respectively. The majority (80-99% +/- 0.8) of plasma radioactivity at all postinjection plasma collection time points was in the form of tyrosine-containing enkephalin metabolites, indicating a substantial and rapid in vivo hydrolysis rate for exogenously administered LE. Leu-enkephalin is metabolized in vivo faster than previously reported in vitro in mouse plasma. However, despite this extensive hydrolysis, levels of intact LE remaining in plasma following its systemic administration are within or above endogenous LE plasma levels.


Regulation of cholecystokinin secretion from a rat medullary thyroid carcinoma cell line: role of calcium, cyclic nucleotides, glucocorticoids, neurotensin, and calcitonin gene-related peptide. CCK-secreting WE rat medullary thyroid carcinoma cell line resembles other calcitonin-producing (C-cell) lines in that calcium, cAMP, or agents which raise cAMP, dexamethasone, and beta-adrenergic agents all stimulate peptide secretion. Unlike other C-cell lines, the WE cells respond similarly to IBMX (3-isobutyl-1-methyl-xanthine, a phosphodiesterase inhibitor) in the presence and absence of forskolin, implying that these cells secrete substances that raise cAMP levels, whose effect is accentuated by IBMX. Both CGRP and neurotensin, peptides that may be secreted by these cells, caused a small, but significant, increase in CCK secretion. It is possible that these or other secreted substances that activate adenylate cyclase are responsible for the cell's high rate of CCK secretion. Their high rate of CCK synthesis and their regulated secretion suggest that these cells will be a good model for studies of CCK expression, biosynthesis, and processing.


Effects of high temperature on growth and efficiency of male and female broilers from lines selected for high weight gain, favorable feed conversion, and high or low fat content. Male and female broiler chicks from five different broiler crosses (WI, LF, and HF = Israeli chicks selected for high body weight gain, and low and high abdominal fat, respectively; FC and WN = Dutch chicks selected for favorable feed conversion and high body weight gain, respectively) were raised at a high ambient temperature (32 to 33 C). Weight gain, protein and fat content in the carcass and feed, and protein efficiency were determined at 4, 6, and 8 wk of age. The effect of the high temperature was evaluated by comparing these data with those of similar chicks raised at a normal temperature (20 to 33 C) up to 6 wk of age. The reductions in body weight, protein gain, and feed and protein efficiency due to the high temperature increased with age and were much larger in males than in females. This trend was more pronounced in WI and WN chicks than in LF, HF, and FC chicks. Females of WI and WN crosses were as heavy as males at 6 wk and heavier at 8 wk. In LF, HF, and FC crosses, both sexes had similar weights at 8 wk. Growth reduction due to the high temperature was largest in the groups with the highest growth rate at the normal temperature (WI and WN males). Chicks with a lower growth rate and a higher capacity for energy storage in fat depots (all females, HF chicks), or a higher capacity for heat loss (FC chicks), were less affected by the high temperature. The results suggest that females should be preferred over males for broiler production in hot facilities or locations. Broiler genotypes selected for feed efficiency at the expense of fast growth may allow for a more profitable broiler production in high-temperature regions.


Differential effects of amylin on memory processing using peripheral and central routes of administration. Amylin is a peptide hormone secreted from the beta cells of the pancreatic islets. Amylin was administered peripherally or centrally following weak or strong training on footshock avoidance conditioning in a T-maze. Under conditions of weak training, amylin improved memory retention in a dose-dependent manner. Under conditions of strong training, it impaired retention over the same dose range. Central administration of amylin in mice given strong training impaired retention but had no effect on the retention of mice given weak training. These findings suggest that the mechanisms of action by which amylin altered memory processing are different for peripheral and central administration. Peripherally secreted amylin may play a role in the amnesia seen in diabetes and the memory enhancement following glucose administration.


Identification of four chicken gastrins, obtained by processing at post-Phe bonds. Chicken antrum was found to contain 7 nmol/g of carboxyamidated gastrin/CCK-like peptides. The predominant chicken gastrin (so named due to the antral origin) contained 53 amino acid residues: DWPEPPSQEQ QQRFISRFLP HVFAELSDRK GFVQGNGAVE ALHDHFYPDW MDF-NH2. Three smaller (less abundant) forms corresponded to the 30-, 21-, and 7-residue carboxyamidated C-terminal fragments. The major part was sulfated at the tyrosine residue in position seven from the C-terminus. A lower isoelectric point and abrupt termination of the sequencing suggest that some of the peptides had an isoAsp-Gly bond instead of an Asn-Gly bond. The three shorter forms were all derived from the precursor by post-Phe cleavages. This cleavage pattern suggests a processing enzyme specific for bonds between Phe and moderately hydrophobic residues.


Hemodynamic effects of calcitonin in the normal rat. Calcitonin (CT) was administered acutely (IV 4-8 U/kg) and chronically (SC 2 U/kg/day x 150 day) to normal male rats. Measurements included heart rate (HR), mean blood pressure (MBP), cardiac index (CI), peripheral vascular resistance (PVR), and stroke volume index (SVI). The MBP was higher in CT rats examined under pentobarbital anesthesia. Upon awakening from anesthesia, rats chronically on CT exhibited impaired recovery of CI and SVI. Hemodynamic effects were not seen in rats acutely treated with CT. Heart weight was unchanged in chronic treatment with CT. Therefore, CT had minimal hemodynamic effects in the normal male rat.


Deoxyribonucleic acid fingerprint bands linked to loci coding for quantitative traits in chickens. Efficacy of DNA fingerprint (DFP) bands in marker-assisted selection programs for quantitative traits in chickens was evaluated. A cross between two populations of White Plymouth Rock chickens that had been selected for 31 generations for high (HW) or low (LW) 8-wk body weight served as the base population for the experiment. Full- and half-sib families were produced over four generations and 400 offspring were measured for body weight at 8 wk (BW8) and shank length at 12 wk of age (SL12). Distributions were constructed for each quantitative trait in offspring of one F1 sire. The DFP produced from mixed blood of the individuals within each tail of the distribution were compared. From a total of 13 DFP bands that were disparate in intensity between the tails, four bands were chosen for analysis. Matings were made between males and females based on the presence or absence of these bands, but were limited to individuals that were within .5 standard deviation of the mean for the distribution of a particular trait. Quantitative traits of the resulting progeny were analyzed to determine whether parental type (presence or absence of the DFP band) influenced expression of the trait in the offspring. One band out of the four tested was associated with SL12, was an effective predictor of phenotype for both SL12 and BW8, and appeared to be inherited in a dominant fashion.


Synthesis and in vitro evaluation of [Leu13]porcine motilin fragments. Several peptide fragments representing N-terminal, C-terminal, and internal sequences of [Leu13]porcine motilin ([Leu13]pMOT) were synthesized using Fmoc solid phase methodology. Peptides were assayed for motilin receptor binding activity in a rabbit antrum smooth muscle preparation and for stimulation of contractile activity in segments of rabbit duodenum. In vitro activity was directly correlated with motilin receptor binding affinity for all [Leu13]pMOT fragments examined. N-Terminal fragments of just over half the length of the native peptide are nearly equipotent as full-length motilin. These results suggest that the N-terminal segment, together with residues from the mid-portion of the molecule, constitutes the bioactive portion of pMOT. The C-terminal segment, in contrast, contributes little to receptor binding affinity or in vitro activity.


A feather-sexed strain of laying hens was more responsive to dietary supplements of choline and methionine than a vent-sexed strain. A response surface design was used to study Cho and Met interactions with corn and soybean diets, using two strains of hens. The strains were a feather-sexed line (FS strain), and a vent-sexed line (SS strain). The diets contained 3% meat and bone meal and, on chemical analysis, 15.1% crude protein, .29% Met, .225% Cys, and 1,041 ppm of Cho. Nine diets were fed from 20 to 68 wk of age, using added Met levels ranging from 0 to 500 ppm and added Cho levels ranging from 0 to 1,500 ppm, to fix the design points. The FS strain consumed significantly more feed per day (117 versus 108 g) than the SS strain, but there were no significant differences for the 24 to 68 wk period in egg production, egg weight, or feed per dozen eggs. Three and five combinations of Met and Cho were significant in improving egg production (P less than .05) out of the eight combinations for the SS and FS strains, respectively. The best egg production for the FS strain for the period 24 to 68 wk was observed at 250 ppm Met and 1,500 ppm Cho, or 427 ppm Met and 220 ppm added Cho. The SS strain showed no significant (P greater than .05) dietary responses in egg production between 250 ppm Met and no Cho, or 427 ppm Met and either 220 or 1,280 ppm Cho. The SS strain showed no significant (P greater than .05) dietary response in egg weight to either Cho or Met.(ABSTRACT TRUNCATED AT 250 WORDS)


The effect of light source and intensity on turkey egg production. The effect of light source and intensity on egg production, fertility, hatchability, egg weight, and shell quality in turkey hens was investigated. Turkey hens were exposed to either incandescent, high-pressure sodium vapor (HPSV), or daylight fluorescent (FL) lights at an intensity of 21.6 1x during the growth phase (after 8 wk). During the breeder phase, hens were assigned to the same light sources at light intensities of either 53.8 or 166.4 1x in a 3 x 3 x 2 factorial arrangement of treatments. Light source during the growth phase had no significant effect on subsequent egg production, egg weight, fertility, or hatchability. Egg production, shell quality, fertility, and hatchability were also unaffected by light source or intensity during the breeder phase. Egg weight was significantly increased in hens exposed to HPSV compared with incandescent lights during the breeder phase. The HPSV and FL lights can be used effectively for turkey breeder hens both during the prebreeder and breeder periods. Furthermore, light intensities of 53.8 and 166.4 1x appear to be equally effective in stimulating production.


Sex differences in the effects of neuropeptide FF and IgG from neuropeptide FF on morphine- and stress-induced analgesia. There is evidence suggesting that the endogenous mammalian octapeptide FLFQPQRFamide (F8Fa or neuropeptide FF, NPFF) has modulatory effects on opioid-mediated analgesia in rodents. There is also substantial evidence for sex differences in opioid analgesia, whereby male rats and mice display greater levels of opioid-mediated analgesia than females. In the present study, determinations were made of the effects of NPFF and IgG from antiserum against NPFF on morphine- and restraint stress-induced opioid analgesia in male and female deer mice. Intracerebroventricular (ICV) administrations of NPFF (0.10-10 micrograms) reduced in a dose-dependent manner morphine- and stress-induced analgesia in both male and female mice, with NPFF having markedly greater antagonistic effects in the male than female mice. Additionally, ICV administrations of NPFF-IgG increased the levels of morphine- and stress-induced analgesia and significantly reduced basal nociceptive sensitivity in male mice, whereas, in female mice, NPFF-IgG had no significant effects on either opioid-mediated analgesia or nociceptive sensitivity. These results indicate that there are sex differences in the modulatory effects of NPFF on opioid-mediated analgesia.


Effects of corticotropin releasing hormone on appetitive behaviors. Corticotropin releasing hormone (CRH) is a 41-residue hypothalamic neuropeptide that has been shown to have potent behavioral effects in animals and has been implicated in clinical disorders in man. This review focuses on those aspects of the behavioral effects of CRH related to food-associated behaviors. The effects of CRH on food intake are compared with its effects on performances maintained by food presentation, and contrasted with the effects of CRH on performances maintained by other events. The effects of CRH antagonists and drugs that interact with the behavioral effects of CRH are also reviewed, particularly with respect to their direct effects on food intake. Lastly, data assessing the effects of CRH administration on central neurotransmitter levels are presented and compared with levels seen in clinical populations. The effect of CRH on food intake seen in animals is consistent with a putative role for CRH in clinical syndromes where appetite suppression is apparent. Since some of the effects of CRH on food intake are subject to pharmacological intervention, strategies directed at peptidergic mechanisms of psychiatric disorders should be explored.


Effect of number of hens per nipple waterer on the performance of several strains of layers in cages. Two experiments were designed to study the effects of housing in cages with several hens per nipple waterer (HPN) ratios on performance of several strains of White Leghorn pullets. In Experiment 1, only body weight gain and water consumption were significantly affected by the HPN; hens at the 2:1 HPN gained more weight and consumed more water per day than those at the 4:1 HPN. The lack of a significant strain by HPN interaction indicated that the four strains responded similarly to the different HPN ratios. In Experiment 2, hens in cages with 3.5:1 and 7:1 HPN consumed significantly more water and feed than those in cages with 10:1 and 14:1 HPN. In both experiments, the HPN had no significant effect on age at sexual maturity, egg production, mortality, and egg weight, but efficiency of feed usage for egg production decreased with the 3.5:1 and 7:1 HPN.


Effect of early feed restriction on growth, feed conversion, and mortality in broiler chickens. Two floor pen and two battery experiments were conducted to determine the effects of early feed restriction on the performance of commercial broilers. Feed restriction was induced in all experiments by providing chicks with 40 kcal of ME per bird per day, commencing at 4 days of age. Male chicks were feed-restricted for 7 (Experiments 1 and 2) or 6 days (Experiments 3 and 4), whereas broiler females were restricted for 5 days (Experiment 1). Ad libitum feeding was resumed after the restriction periods, and continued through the conclusion of the experiments at 49 (Experiments 1 and 2) or 28 (Experiments 3 and 4) days of age. Broilers provided ad libitum access to feed for the entire experimental period served as the controls in each study. Broilers subjected to an early feed restriction had significantly (p less than or equal to .05) lower mean body weights than controls for all ages measured in the four experiments. However, feed conversion ratios for restricted broilers were significantly lower at 28 (Experiments 1 through 4) and 49 (Experiments 1 and 2) days of age than for birds consuming feed ad libitum. Through regression analyses, it was estimated that male broilers in Experiments 1 and 2 would require approximately 2 additional days to obtain body weights similar to those observed in control broilers and would still maintain a lower feed conversion ratio at this older age. In Experiments 1 and 2, weekly body weight gains for restricted broilers were significantly lower than for controls from 0 through 28 days of age. However, restricted broilers (7 days) in Experiment 2 had significantly higher rates of gain from 29 to 49 days of age than unrestricted controls. Total pen body weights for restricted and ad libitum groups were similar at 49 days of age in Experiments 1 and 2, which reflected the significant difference in mortality observed between the two groups.


Issues facing researchers.

Issues facing researchers. Agriculture research spans the spectrum from fundamental to applied research. Researchers in agricultural fields must compete successfully with scientists in their field of research for funding during a time of diminishing resources. In addition, there are pressures on agricultural researchers within their institutions and at the state and national levels that often differ from pressures on scientists in fields other than agriculture. In order to be successful, the poultry scientist must compete for funds, keep current in a rapidly advancing technological environment, and attempt to meet the needs of the poultry industry.


Legislative view of research.

Legislative view of research. Congress has its own perception of research; whether it equates to reality is up to the research community. Many in Congress perceive that the research community is free-floating and is not focused on addressing the problems that face the United States today. Recent government reports have reviewed the purposes, accountability, and challenges facing the traditional research and extension system as it enters the 1990s. These reports send a strong message that the status quo will not work sufficiently in the 1990s. How does this affect agricultural research dollars? Congress, in response to its own perceptions, has strived to establish priorities by allocating resources to key projects. Whether this is in the best interest of the nation is debatable. The agricultural research community needs to establish priorities and to communicate them effectively; otherwise someone less informed in the Congress will do it.


Oocysts of Eimeria in the litter of broilers reared to eight weeks of age before and after withdrawal of lasalocid or salinomycin. Forty-six broiler houses were examined for the presence of oocysts of Eimeria in the litter before and after withdrawal of lasalocid or salinomycin from the feed. A decrease in number of small oocysts (Eimeria acervulina or Eimeria mitis) was observed following withdrawal of medication. Numbers of medium-sized oocysts (probably Eimeria tenella) remained the same, but an increase in large oocysts (Eimeria maxima) was recorded. No lesions attributable to Eimeria tenella were found in the ceca of birds after withdrawal of the drug. More small- and medium-sized oocysts were found at sites with new litter than at sites where the litter had been employed for previous flocks. No difference in the number of oocysts was found, whether birds were reared on oak shavings or a mixture of pine shavings and rice hulls. The number of oocysts was positively correlated with the moisture content of the litter, but there was no correlation between oocysts present and the final BW or feed conversion of the birds. Moisture levels were highest (after withdrawal of drug) for new litter or oak shavings. There was no correlation between moisture content of the litter and BW or feed conversion.


Effect of dietary calcium level on medullary bone calcium reserves and shell weight of Leghorn hens. Two experiments were conducted to determine the effect of different levels of dietary Ca and subsequent feeding of a very low level of Ca on the medullary bone Ca reserves of laying hens. In Experiment 1, a total of 30 40-wk-old Single Comb White Leghorn hens were offered a diet with 2.5, 3.5, or 4.5% Ca for a period of 21 days with 10 birds per dietary treatment. On Day 21, five hens from each dietary treatment were euthanatized. The remaining birds were offered a .46% Ca diet for 5 days. In Experiment 2, 25 Single Comb White Leghorn hens were fed a 3.5% Ca diet for 21 days. Hens were then offered a .46% Ca diet and five hens euthanatized on Days 0, 2, 3, 5, and 7 of feeding the low-Ca diet. In Experiment 1, dietary Ca level had a significant (P less than .05) effect on total medullary Ca reserves of laying hens. Previous dietary Ca level had no significant (P greater than .05) effect upon medullary bone Ca reserves after subsequently feeding the low-Ca diet for 5 days. There was a (P less than .05) significant reduction in medullary bone Ca reserves of hens, regardless of previous level of calcium fed. In Experiment 2, feeding a low level of dietary Ca resulted in a significant (P less than .01) reduction in medullary bone Ca reserves of all bones measured, except the humerus. Although there was a significant (P less than .05) reduction in medullary bone Ca during the depletion period, hens appeared to make some attempt to conserve medullary bone Ca reserves.


Dietary poultry fat and gastrointestinal transit time of feed and fat utilization in broiler chickens. An experiment was conducted with broiler chicks to determine the effect of 0, 5, 10, and 20% supplemental poultry fat and age on gastrointestinal transit time (GTT) and the effect of supplemental fat on fat utilization and growth. Mean GTT, measured with chromic oxide or ferric oxide, was not affected by supplemental fat. There was a curvilinear relationship (rising ogive) between mean GTT and age. It increased from an estimated lower plateau of 170 min to an upper plateau of 211 min with the inflexion point at 3.23 wk. At 6 wk of age, birds receiving supplemental fat consumed more energy and were heavier and more efficient. Total lipid digestibility increased with supplemental poultry fat but digestibility of poultry fat was not altered. The AME of poultry fat ranged from 8.1 to 8.4 kcal/g at 5 to 20% inclusion in the diet.


Effects of protein and amino acid level fed zero to forty-two days on response of broilers to protein and amino acid levels fed forty-two to forty-nine days of age. Three studies were conducted to evaluate the effect of protein and amino acid (AA) levels fed 0 to 42 days posthatch on response of broiler chickens to levels of protein and AA fed during the finisher period. Birds were grown to 42 days on diets formulated to contain a minimum of 105 or 120% of recommended AA levels, and then randomly assigned to diets formulated to contain a minimum of 70, 80, 90, 100, 110, or 120% of recommended AA levels for 42 to 49 days. No minimum protein level was imposed on the diets. Body weight, feed intake, feed utilization, dressing percentage, abdominal fat content, and parts yield were determined. The protein and AA level fed 0 to 42 days had significant effects on 42- and 49-day body weight and 42- to 49-day feed intake of male but not female broilers. However, there was no interaction of protein and AA levels fed 0 to 42 days on response to protein and AA levels fed 42 to 49 days for these criteria. Weight gain, feed intake, and feed utilization of both males and females 42 to 49 days were significantly influenced by protein and AA levels fed during this time; protein and AA requirements of females during this time appeared to be equal to, or greater than, those of the male broiler. Although there were significant interactions of protein and AA levels 0 to 42 and 42 to 49 days on various carcass parameters for both male and female broilers, these followed no consistent pattern. Protein and amino acid levels fed 0 to 42 days had a significant influence on both quantity and percentage of abdominal fat in both male and female broilers processed at 49 days but not on any other carcass parameters. The protein and AA levels fed 42 to 49 days had a significant effect on both quantity and percentage of abdominal fat in male but not female broilers. The protein and AA levels fed 42 to 49 days significantly influenced quantity but not percentage of breast meat in male but not female broilers. The results of the current study indicate that there was minimal carryover effect of protein and AA levels fed 0 to 42 days on response to AA levels fed 42 to 49 days.


Relationship of hen age and egg sequence position with fertility, hatchability, viability, and preincubation embryonic development in broiler breeders. Indian River broiler breeder hens (n = 29) were caged individually to investigate whether hen age and egg sequence position were related significantly to the dependent variables fertility, hatchability, viability (hatch of fertile eggs), and preincubation embryonic development. Hens were artificially inseminated once per week. All eggs laid during the period of 31 to 54 wk of age were stored at 16 to 17 C for .5 to 7 days. Time of oviposition records were used to assign eggs to sequence position ("first" or "subsequent"). Eggs laid on odd-numbered weeks were broken open, fertility determined, and embryonic development staged. Eggs laid on even-numbered weeks were sent to a commercial hatchery to assess hatchability. Unhatched eggs were opened to determine fertility and embryonic mortality. In addition, hen weight, number of days since insemination, time of oviposition, and egg weight were recorded to determine their relationship to the dependent variables. Fertility (n = 3,240 eggs) and hatchability (n = 1,653 eggs) were not significantly related to egg sequence position, but were related to hen age (P = .0001 and P = .0002, respectively). Older hens demonstrated lower fertility and hatchability. In contrast, embryo viability (n = 1,487 eggs) and preincubation embryonic development (n = 1,200 eggs) were not significantly related to hen age, but were related to egg sequence position (P = .0026 and P = .0001, respectively). First-of-sequence eggs had lower viability, and embryos of these eggs were more developed than embryos of subsequent eggs. These data indicate that the reduction in chick production observed as the hen ages may be due to the increased incidence of first-of-sequence eggs.


The effectiveness of ligating or detaching ceca as an alternative to cecectomy. Cecectomy (the surgical removal of ceca) is a technique that has been used to explore the nutritional and osmoregulatory role of the ceca in domestic fowl and other birds. The operation imposes a high level of physiological stress on a bird. The current study describes the effectiveness of ligating or detaching the ceca in young turkeys as an alternative to complete removal of the organ. Birds with ligated ceca showed less postsurgical depression and initiated feeding and other normal activities sooner than birds with detached ceca. Birds with detached ceca also had a significantly slower rate of weight gain 1 wk after surgery. Both surgically treated groups were more depressed than were sham-operated turkeys. Post-mortem examinations revealed that cecal detachment was more successful in actually separating the ceca from the rectum and in preventing flow of digesta in and out of the ceca. Evidence of growth of cecal stumps that remained attached to the rectum was found following both ligation and detachment. Post-mortem examinations revealed that in four birds both ligated or detached ceca left in the abdominal cavity became enlarged and filled with a dark, pasty, odorous substance. This increase in cecal size and volume of contents with time apparently indicates that some microbial metabolic activity continued in the cecal lumen. If so, this might have negative implications for studies that assume that such ceca are no longer functional.


A rapid method for the determination of temperature abuse of fresh broiler chicken. Ready-to-cook broiler chicken carcasses were obtained immediately postchill from a commercial processing plant. A control group was held at 4 C for 10 days. Test groups were held at 4 C for 2 days, temperature abused at 10, 15, or 20 C for 0, 1, 2, 4, 6, 8, 12, or 24 h, and then held at 4 C for an additional 4 days. Both control and test birds were sampled using whole carcass rinse techniques and bacteria enumerated using impedance detection times (DT) and total plate count (TPC) techniques at incubation temperatures of 18 and 42 C. The DT and TPC showed a significant increase (P less than .05) in the number of organisms recovered at 18 C but did not show a significant increase (P less than .05) in organisms recovered at 42 C when carcasses were held at 4 C continuously for up to 10 days. Carcasses that were temperature abused at 10 C for up to 24 h were not significantly different (P less than .05) from the unabused controls, using either DT or TPC, following incubation at 18 and 42 C. At an incubation temperature of 18 C, carcasses abused at 15 or 20 C had significantly more growth of bacteria than controls at 1, 6, 12, and 24 h and 4, 8, and 12 h as determined by DT and at 6, 12, and 24 h and 1, 2, 4, 8, and 12 h, respectively, as determined by TPC.(ABSTRACT TRUNCATED AT 250 WORDS)


Research note: open-field behavior of Japanese quail chicks genetically selected for low or high plasma corticosterone response to immobilization stress. Open-field behavior was examined in Japanese quail chicks genetically selected for either reduced (LS, low stress) or exaggerated (HS, high stress) plasma corticosterone response to immobilization stress. Chicks of the LS line showed less freezing and ambulated sooner than did their HS counterparts. These findings suggest that exposure to an open-field or novel environment elicited less fear in LS than in HS chicks. They also support the suggestion that fearfulness and adrenocortical activation are positively associated and indicate that selection for differential adrenocortical responsiveness exerted concomitant effects on fear-related behavior.


Research note: maintenance of duodenum weight during a molt induced by dietary zinc in a low-calcium diet. The duodenal loop was excised from hens that had been induced to molt by a low-calcium diet containing 2,800 ppm zinc in the form of zinc sulfate heptahydrate. This was compared to that of hens pair-fed a layer diet. In Experiment 1, hens exhibited an increased duodenum weight after receiving the zinc diet for 14 days but no differences remained 14 days later. In Experiment 2, increased duodenum weight was noted after 4 days on the zinc diet and this persisted through 10 days. This greater tissue weight was observed whether expressed on a wet or dry weight basis. Thus, there was a persistency of tissue mass. The duodenum does not regress during a zinc-induced molt as it does during a fast-induced molt.