The isolation and characterization of bacteriophage T7 messenger RNA fragments containing an RNase III cleavage site. We have isolated overlapping RNA fragments which contain the region surrounding the ribonuclease III cleavage site between bacteriophage T7 genes 0.3 and 0.7. Although all of these fragments contain the site of cleavage, only certain fragments are correctly recognized and cleaved by RNase III. Analysis of the cleavage products of the fragments indicates that the enzyme produces a single endonucleolytic break at this site in the T7 early RNA precursor molecule. In addition, the 3'-terminal adenylic acid residues observed previously on the in vivo T7 early RNA species were not found in these fragments and, therefore, must represent a post-transcriptional, post-processing modification of the RNA.


Preparation of 2'-thio-2'-deoxycytidine 2':3'-phosphorothioate. The synthesis of a novel ribonucleotide analog 2'-thio-2' deoxycytidine 2':3'-O,S-phosphorothioate is described. In the first step, 2,2'-anhydro 1-beta-D-arabinosylcytosine was thiophosphorylated by the action of dithiophosphate, a process which gave predominantly the 3'-O-phosphorothioate isomer. An intramolecular displacement reaction led to the formation of the title compound. Structure and reactivity of this thioanalog differ substantially from 2':3'-CMP.


Conformations of satellite DNAs.

Conformations of satellite DNAs. X-ray fiber diffraction studies of satellite DNAs from Gecarcinus lateralis, Drosophila virilis and Mus musculus, all of which have highly repetitious base sequences but with different degrees of sequence complexity, reveal only classical polynucleotide duplex structures in contrast to some highly repetitious synthetic DNAs.


Models of triple-stranded polynucleotides with optimised stereochemistry. Detailed models are presented for the triple-stranded polynucleotide helices of poly (U)-poly (A)-poly (U) (two forms), poly (U)-poly d (A) -poly (U), poly d(C)-poly d(I)-poly d(C), poly d(T)-polyd(A)-poly d(T) and poly (I)-poly (A)-poly (I). The models were genrated using a computerized, linked-atom procedure which preserves standard bond lengths, bond anglesand sugar ring conformations, constrains the helices to have the pitches and symmetries observed in X-ray diffraction experiments, and optimises the non-bonded interatomic contacts including hydrogen bonds. The possible biological sigificance of such complexes is discussed.


Sequence organization of porcine DNA.

Sequence organization of porcine DNA. The sequence organization of porcine DNA isolated from thyroid has been analyzed by hydroxylapatite (HAP) chromatography. The reassociation of 0.4 kilobase (Kb) DNA fragments shows, besides the presence of 5% inverted repeat sequences (foldback DNA), that 45% of the genome is represented by high (10%) and intermediate (35%) repetitive components, whereas the remaining 50% is unique sequences. 30% of the unique sequences consists of 1,000 nucleotide fragments interspersed with repetitive elements 400 nucleotides in length. The remaining 20% is longer unique sequences (10,000 nucleotides) apparently not linked to repetitive elements.


Circular dichroism as a probe of DNA structure inside reconstituted nucleohistones. Reconstituted nucleohistones were obtained by mixing in given conditions acid extracted histones and eukaryotic DNA. The histone/DNA ratio (w/w) was in the range 0.35 - 0.95. With the four histones (H2A2B) we have been able to obtain subunits (nucleosomes or upsilon-bodies). The variation of cirsular dichroism signal with temperature at 280 nm was measured to follow structural changes of the DNA inside the complex. The true change of ellipticity (see article) of histone-bound DNA regions, is similar for reconstituted nucleohistone and H1-depleted chromatin, and is therefore a physical probe of the presence of nucleosomes.


The effect of (2'-5') and (3'-5') phosphodiester linkages on conformational and stacking properties of cytidylyl-cytidine in aqueous solution. Conformational properties of (2'-5') and (3'-5') CpC have been determined by proton magnetic resonance spectroscopy at 220 MHz. The ribose ring structures are predominantly 3E with the exception of the ring from the 2'-phosphate fragment of C(2'-5')pC which exhibits an 2E pucker. Bases are oriented anti with respect to the ribose and the conformations about C4'-C5', C5'-O5', C3'-O3' (C2'-O2') are gg, g'g', and g+ in equilibrium g-, respectively. The dimers exist as mixtures of stacked (g+g+ and g-g- about the P-O(C) bonds) and unstacked species at 20 degrees C. Stacking is estimated to be 35% in both dimers.


Preparative fractionation of DNA restriction fragments by high pressure column chromatography on RPC-5. High pressure liquid chromatography on the RPC-5 reversed-phase ion exchange system has been shown to have several potential applications as an initial high capacity step in the isolation of specific DNA restriction fragments. The fractionation of the Hinc II digest of lambda DNA, which contains 35 fragments with "flush ends" ranging in size from 3 x 10(6) to 7 x 10(4) daltons, has been used as a model system. Under certain conditions there are some restriction fragments whose elution relative to other fragments is different on RPC-5 chromatography than it is on gel electrophoresis. In some special circumstances it is possible to obtain satisfactory yields (60-70%) of a pure restriction fragment after a single passage through an RPC-5 column.


Cleavage of adeno-associated virus DNA with Sali,Psti and Haeii restriction endonucleases. Duplex AAV-2 DNA was digested with SalI, PstI or HaeII restriction endonucleases and the cleavage sites were mapped. SalI cleaves AAV DNA at 0.310 map units, PstI at 0.106, 0.422 and 0.914 and the five HaeII sites were mapped at 0.110. 0.156, 0.181, 0.536 and 0.600 map units. These cleavage products will be useful for the isolation of specific regions from the AAV DNA, located outside of the stably transcribed region of the genome, and will also help to map more complex restriction enzyme cleavages.


Nuclease digestion in between and within nucleosomes. In the course of digestions of rat liver nuclei with micrococcal nuclease the size of the nucleosomal DNA is shortened by 50-60 nucleotide pairs from starting lengths of about 200, 400, 600, 800, etc. nucleotide pairs in the monomeric and oligomeric nucleosomes, respectively. Acid soluble DNA material is created relatively slowly as compared to the rate of formation of subnucleosomal material. More DNA with lengths in between the 200, 400, etc. nucleotide pairs of nucleosomal DNA is formed when digestions with micrococcal nuclease are carried out at 0 to 10 degrees C compared to 40 degrees C. With DNAase II, on the other hand, formation of a 200 nucleotide pair pattern is favoured at the low temperatures. Apparently, the accessibility of potential cleavage sites in between and within nucleosomes depends strongly on the conditions of digestion. Possible reasons for this dependence are discussed.


Isolation of native DNA fragments containing structural genes at the beginning, in the middle or at the end of the coding strand. A method for the isolation of the DNA fragments containing structural genes at the beginning, in the middle and at the end of the coding strand is described. It involves mechanical or enzymatic shearing, exonuclease treatment, hybridization with mRNA and subsequent retention of DNA-mRNA complex on poly(U)-Sepharose. Hybridization of the DNA fragments with mRNA takes place with high specificity and isolated material is enriched in structural genes. Applications of this method for gene isolation are discussed.


A possible structure for calf satellite DNA I. Calf satellite DNA I (p = 1.715) has been hydrolysed by a number or restriction endonucleases. It consists of a repeating unit of 1460 nucleotide pairs within which the sites of Eco R II Mbo I, Sac I, Alu I, Ava II and Hha I were localised in comparison with those of Eco R I and Hind II. The distribution of the Hpa II, Sac I, Hha I, Hinf I and Mbo II sites within calf satellite DNA I, as well as that of several restriction endonuclease sites within calf satellite DNA III (p = 1.705) allowed me to define subsatellite fractions. Furthermore, some of the sites of the CpG containing restriction enzymes Hpa II and Hha I are lacking. The possible implications of these results are discussed.


Topographical analysis of yeast ribosomal DNA by cleavage with restriction endonucleases. II. The physical map of EcoRI fragments. Yeast ribosomal DNA (rDNA) was digested with the restriction endonuclease EcoRI. Eight distinct fragments were obtained with a molecular weight of 4.35 (1), 1.75 (2), 1.45 (3), 1.07 (4), 0.42 (5), 0.37 (6), 0.26 (7) and 0.22 x 10(6) (8) daltons, respectively. Except for fragment 1 with a molecular weight of 4.35 x 10(6) daltons, all fragments are derived from the multiple ribosomal transcription units. The 'spacer' sequences, on the other hand, gave rise to digestion products which are very heterogeneous in size. By analysis of the partial digestion products which are very heterogeneous in size. By analysis of the partial digestion products, together with the data obtained by digestion with a combination of two restriction enzymes (EcoRI and Hind II or Hind III) and redigestion of the Hind II-and Hind III-fragments with EcoRI, the physical map of the EcoRI cleavage sites in the ribosomal transcription unit could be established.


Synthesis of 3'-phosphates of diribonucleoside monophosphates via phosphotriester intermediates. Phosphorylation of the easily accessible 3',5'-diesters 1a-d with diphenyl phosphorochloridate, followed by selective 5'-deacylation, affords the phosphotriester derivatives 2a-d in good yields. Alkaline treatment of 2a-d results in the formation of the 2',3'-cyclic phosphates (3a-d). The usefulness of the phosphotriester derivatives 2a-d is also demonstrated in the synthesis of the nucleotidyl-(3'-5')nucleoside 3'-phosphates U-Up (10a), U-Ap (11a), U-Cp (12a) and A-Gp (13a). The fully protected dinucleoside diphosphates 5c-8c, prepared by the phosphotriester method, are deprotected in two ways: (a) by a purely chemical method, affording the dinucleoside diphosphates in a circa one to one mixture of 2'- and 3'- isomers, 10b-13b and 10a-13a, respectively, and (b) by a mixed chemical-enzymatical approach which gives the pure 3'-phosphates (10a-13a).


Interspersion of different repeated sequences in the wheat genome revealed by interspecies DNA/DNA hybridisation. The repeated sequences in oats DNA have been used to study chromosomal repeated sequence organisation in wheat. Approximately 75% of the wheat genome consists of repeated sequences but only approximately 20% will form heteroduplexes with repeated sequences from oats DNA at 60 degrees C in 0.18 M Na+. The proportion of wheat DNA that forms heteroduplexes with oats DNA is shown to be independent of the wheat DNA fragment length. However, the proportion of wheat DNA that is retained with the heteroduplexes when fractionated on hydroxyapatite is very dependent upon the wheat fragment length up to 3500 nucleotides. This is because more non-renatured wheat DNA is attached to the heteroduplexes with longer fragments. The results indicate that the repeated sequences in the wheat genome homologous to repeated sequences in oats are not clustered in the chromosomes but distributed amongst other repeated and possible non-repeated sequences.


The nucleotide sequence of a DNA fragment, 71 base pairs in length, near the origin of DNA replication of bacteriophage 0X174. Part of the nucleotide sequence of a restriction fragment covering the origin of phiX174 DNA replication 1 has been determined. The fragment A7c was obtained by digestion of phiX174 RF DNA by the restriction enzyme from Arthrobacter luteus, Alu 1. It was further cleaved into two fragments, one large and one small, by the action of the restriction enzyme from Haemophilus aegyptius, Hae 111. The nucleotide sequence of the small fragment has been determined by analysis of the transcription products obtained by the action of Escherichia coli DNA-dependent RNA polymerase on denaturated template under conditions of low salt. Transcripts longer than the template were found. The whole sequence of 71 nucleotide pairs could be derived from complementary oligonucleotides, obtained after digestion of the transcripts with T1 or pancreatic RNAase. The sequence suggests that at least 4 of the 5 amber mutants 2 that have been mapped on this fragment are identical. On account of this and other evidence a reading frame is proposed.


Incorporation of 5-substituted uracil derivatives into nucleic acids. Part IV. The synthesis of 5-ethynyluracil. 5-Acetyluracil (I) has been treated with POCI3 to give 5-(1-chlorovinyl)-2,4-dichloropyrimidine (II). Treatment of II with KOEt gave a mixture of 2-ethoxy-5-ethynyl-4 (3H)-pyrimidinone (IIIA) and 4-ethoxy-5-ethynyl-2 (1H)-pyrimidinone (IIIB). IIIA and IIIB were isolated and characterised. The mixture of IIIA and IIIB upon treatment with HCI gave 5(1-chlorovinyl)uracil (IV). Reaction of IV with KOEt gave 5-ethynyluracil (V). 5-Ethynyluracil was more easily obtained by the treatment of II with KOH in aqueous dioxan.


Induction of interferon by polyribonucleotides containing thiopyrimidines. The influence of thioketo substitution in pyrimidine bases of double-stranded polynucleotides on interferon induction was investigated. The stabilizing effect of 2-thioketo substitution was reflected in the increased interferon inducing activity of poly(A-s2U) over that of poly(A-U). Poly(A-s2U) and poly(I)-poly(s2C) were as effective as poly(I)-Poly(C) in rabbit cells. Poly(I)-poly(C) and poly(I)-poly(s2C) were compared in several animal species. No differences in biological effects were observed in rabbits and dogs. In rodents, poly(I)-poly(s2C) was less effective and less toxic.Poly(I)-poly(s2C) was highly resistant against degradation by human serum. Further investigations seem to be justified to elucidate whether this property offers any advantages for the potential clinical utilization of poly(I)-poly(s2C).


Choice of surgical method in treatment of chronic gastric and duodenal ulcer. In the light of an analysis of late results of surgical treatment obtained in a group of 2450 patients with chronic gastric or duodenal ulcer subjected to various types of operations the author stresses the importance of choice of a proper surgical method. It is concluded that in treatment of chronic gastric ulcer the most appropriate method is partial gastrectomy including the whole pyloric part of the stomach. On the other hand, in chronic duodenal ulcer gastrectomy must be more extensive including over 70% of the gastric surface, it may be less extensive if associated with vagotomy. Truncal vagotomy with pyloroplasty should be applied only in high-risk patients in view of a high proportion of recurrences of peptic ulcer.


Hiatus hernias.

Hiatus hernias. In the years 1950--1974 53 cases of hiatus hernia were treated surgically at the Department I of Surgery, Institute of Surgery, Medical Academy in Poznan. These included 32 sliding hernias, 16 paraesophageal, 2 mixed, 1 with acquired short esophagus and 2 with congenital short esophagus. In 17 cases the approach was through the chest, in 35 through the abdominal cavity and in 1 through the chest and abdominal cavity. Late results were favorable.


Congenital chest deformities.

Congenital chest deformities. The authors discuss 334 patients operated upon for congenital deformities of the chest. The clinical manifestations, radiological and electrocardiographic investigations and spirometric measurements as well as postoperative complications are described. The results of follow-up examinations in a part of this material are reported.


Psychopathology of posttraumatic epilepsy in the light of personal clinical observation. The authors present the result of psychopathological analysis of a group of 91 patients who suffered from epileptic seizures of definitely or probably established posttraumatic etiology. Mental disturbances in these patients were differentiated into sporadic psychopathological phenomena and chronic psychiatric syndromes. Sporadic episodes, which the authors identify with "psychical seizures", concerned 58 patients. They were manifested by psychomotor or psychosensory seizures, dismnesia and disthymia. Chronic psycho-organic syndromes were diagnosed in 85 patients. They were: dementive, characteropathic, dementive-characteropathic, oligophrenic and temporal lobe syndrome (according to Landolt). Psychopathological picture of posstraumatic epilepsy does not appear to be specific for the etiological factor. The relatively high number of patients with partial seizures of mixed symptomatology may indicate that head traumas frequently damage the limbic structures of the brain. Confirmed is the observation of other authors that there is a correlation between the temporal localization of epileptic focus and character disturbances.


Clinical evaluation of Lecyvit (POLFA).

Clinical evaluation of Lecyvit (POLFA). Lecyvit POLFA was applied in 114 persons as a roborant drug. Satisfactory results of treatment were observed in 106 subjects (93%). Clinical improvement consisted in increased physical and mental activities, better appetite and gain of body weight.


Renal glomerular and tubular lesions in rabbits treated with uranyl nitrate. Renal injury and regeneration was followed light--and electron microscopically in rabbits treated with uranyl nitrate. The presence and importance of glomerular lesions is noteworthy.


Sensitivity of regenerating rat renal tubular epithelium to mercury bichloride. The purpose of present study was to evaluate the sensitivity of regenerating epithelium of renal proximal convolutions to mercury bichloride and to describe light microscopic, electron microscopic and histochemical abnormalities occurring under the effect of the latter. The ligation of rat kidney with silk thread resulted in ischemic necrosis of tubules of compressed area followed by vivid regeneration. In that period mercury bichloride was administered intraperitoneally. The first detectable lesions appeared in mitochondrial swelling, condensation of endoplasmic reticulum, a decreased number of ribosomes and the presence of numerous vocuoles. Later on, the regenerating epithelium lost its brush broder and single cells became necrotic. During 42 days of observation the sequelae of mercuric intoxication were only focal and particular cells showed injury of variable degree.


Immunological and patho-clinical observations of a patient with rectal carcinoma after long term administration of thymic extract (TFX). TFX (Thymus Factor X) extract from calf thymuses, was given for the first time in Poland to the patient suffering from advanced rectal cancer with perforation to the peritoneal cavity, diffuse fecal peritonitis and multiple metastases to the liver. The immunological, clinical and histopathological observations of the patient for the period of 12 months of treatment are presented. The patient was given the biologically active thymus extract in the total amount of 14.2 mg in 34 subcutaneous injections initially in single dose of 0.1 mg and later on in amount of 0.5 mg. The remarkable restoration of the immunological reactivity of the patient from a total energy to the state of hyperreactivity has to be emphasized. It has been confirmed by the delayed hypersensitivity skin tests (Rt-23, PHA), the increase of the number of RFC (TEa and TEt rosette test with the SRBC), the diminishing of the autolymphocytotoxicity of the peripheral lymphocytes (evaluated by DTL) and also by the high level of the absolute number of circulating lymphocytes in the peripheral blood. The improvement of the clinical condition of the patient consisted in total regression of the inflammatory and functional changes in the respiratory and cardiovascular systems and also in the renal function. The regular rectal endoscopic examinations of the patient revealed the diminished size of the tumor and its unusual appearance described as an induration and sharp delimitation from the surrounding tissues. The histological examinations performed 5 times during the observation period of 11 months revaled the essential changes in the structure of the growth: the marked increase of the stroma with the signs of active proliferation, foci calcification and ossification. These changes were interpreted as extremely rare in the rectal cancer stroma and are considered to represent dystrophic calcification at the neoplastic foci followed by ossification. It should be also pointed out that twelve months survival of the patient with advanced rectal cancer and multiple metastases to the liver represents an unusually long rate in comparison to that generally accepted in the literature for this grade of rectal cancer advancement.


Initial observations on the administration of thymus extract (TFX) in patients with alimentary tract neoplasms. The administration of thymus extract (TFX) has been performed in 35 patients suffering from malignancies of the alimentary tract. TFX, being a calf thymuses extract prepared by Jelenia Góra Drug Producers "Polfa" was given in the dose of 0.5 mg in 3 or 7-day intervals during several months. The immunological condition of patients was checked by many tests: E-rosettes, DLT (direct lymphocytotoxicity test) determination of absolute number of peripheral blood lymphocytes and skin tests (RT-23, PHA and TFX). Our observations revealed that TFX is biologically active also in humans, increases cell-mediated immunity and may restore the immune capability of patients operated due to malignancies of the alimentary tract.


Relationship between the blood glucagon, growth hormone and glucose levels in myocardial infarction. The blood levels of glucagon, growth hormone and glucose were investigated in early (1st day) and late stages (3rd week) of myocardial infarction in 11 cases of severe and 6 cases of mild myocardial infarction. The results were related to those in 9 healthy controls. In severe myocardial infarction associated with complications the glucagon level was statistically significantly higher than in controls in the early as well as in the late stage of the disease. In mild myocardial infarction this was not observed. The growth hormone level was significantly higher than normal in patients with severe myocardial infarction in only a few cases and only in the early stage. The differences between both stages of infarction and between patients and controls were not significant. Blood glucose level in the early stage of severe myocardial infarction was higher than normal and statistically significantly higher than in the late stage. In view of the differences in blood glucose and glucogon levels during severe myocardial infarction it seems that hyperglycemia in early stages of the disease depends on the high catecholamines level in the blood and on the insulin: glucagon index and not on the absolute concentration of glucagon in the blood.


Analysis of familial, somatic and psychic aspects in children with simple obesity. The analyzed material included 60 children aged 5--16 years with the diagnosis of simple obesity. It was found that 30% of them, nearly all girls, had obese parents. This may suggest an increased tendency for familial occurence of obesity in girls. The birth weight of obese children was usually higher than that of children with normal body weight, and 36,6% of children began to show overweight before the age of 3 years. It was found that the index of slender body builds is much lower in obese children than in controls. The analysis demonstrated also that 28.9% of obese children were more or less retarded mentally which may be also a pathogenetic factor in obesity.


Surgical treatment of obesity.

Surgical treatment of obesity. Thirty-two patients, 28 women and 4 men aged from 17 to 57 years were treated surgically for obesity. Their body weight ranged from 90 to 184 kg, with an excess weight from 42% to 180%. In 6 cases the Salmon operation and in 26 the Payne-de Wind operation were performed. Two patients died. In the remaining cases the postoperative course was uneventful and relatively mild. The longest follow-up period was 22 months. Weight loss rate was highest in the first period after the operation and the monthly rate of the weight loss in the first year after the operation was 3.5 to 6 kg. Diarrhea disappeared usually after 4 months. In some patients a transient fall in serum potassium and calcium level was observed. Other favorable results included a significant fall in the serum levels of cholesterol and beta-lipoproteins and improved value of BSP retention test. The condition of the patients is good and they have returned to work.


"Adenomatoid tumor" of female genital tract. Three cases of adenomatoid tumor of the uterus in women aged 30, 40 and 49 years, and three cases of the tumor situated in the oviducts in women aged 39, 43 and 50 years were analyzed micro- and macroscopically. A tubulo-canalicular, a microcystic and a mixed form were distinguished. Inflammatory changes in the oviducts, myomas and myomatosis of the uterus seem to be conducive to growth of adenomatoid tumor. The authors emphasize the diversity of microscopic structure, creating difficulties of differential diagnosis. Some histochemical reactions may be useful in diagnosis.


Blastic transformation of blood lymphocytes in patients with laryngeal carcinoma. In 40 patients with carcinoma of the larynx transformation of lymphocytes under the influence of PHA was studied. In 20 patients, in addition, venous blood flowing from the tumor, collected during the operation was also examined. In 33 patients the index of blastic transformation was lowered as compared with that in the control group. In 15 of 20 patients, the lymphocytes of venous blood from the vicinity of the tumor showed significantly higher indices of blastic transformation in comparision with those from peripheral blood. It was assumed that difference was due to an increased participation of T-lymphocytes in venous blood flowing from the tumor area. T-lymphocytes might move into the blood from the lymphoid infiltrates which accompany the neoplastic tissue.


Histological and electrophysiological changes of the lower motor neurone with aging. Maximal motor conduction velocity of ulnar nerve was measured in 7 patients 80-93 years old, who suffered from cerebro-vascular stroke. In 2 cases motor conduction velocity was significantly decreased. Specimens for histopathological study were taken from the spinal cord at the C8 and Th1 segments and from the ulnar nerve at the elbow level on the side opposite to the hemiparesis. In 100 sections of each senile case and in 3 control cases of younger individuals the motoneurons of the retro-postero-lateral nucleus with visible nucleoli were counted. In the ulnar nerve qualitative and quantitative examinations were performed. The study showed a significant loss of motoneurons in the nucleus and a loss of myelinated fibers especially the thickest ones in the ulnar nerve in elderly cases. In teased preparations axonal degeneration and segmental demyelination of some fibers were seen. Morphological changes in some cases, resulted in a slowed down motor conduction velocity.


Fine structure of human fetal nerve.

Fine structure of human fetal nerve. Sciatic nerve of 16, 18, 20 and 24 weeks old fetuses was examined electron microscope. Morphologic specificiity and difference between the structure of fetal and mature nerves were demonstrated. The myelination of human sciatic nerve was found to begin after the 16th week of fetal life. From that time the number of myelinated fibers increases progressively and the number of fetal fibers decreases. The time of the end of myelination in man is unknown. The studies of the later period of development and maturation of fetal nerve may give necessary information about the period of the end of myelination. In view of increasing interest in pathogensis of comparable data studies on embryogenesis of human peripheral nerve are indispensable for evaluation of some changes observed in congenital diseases. The ultrastructural observations of human fetal nerve are scarce (3, 6, 7, 10) and they concern only single nerves. The morphological specificity of fetal nerve and characteristic difference between fetal and mature nerves require basic data on different forms of fetal nerve and time of transition of one form into another.


Dimensions and geometry of the temporomandibular joint and masseter muscles. The bio-engineering team presents its suggestion of a method for the measurement of the temporomandibular joint and masseter muscles in order to determine the parameters necessary for exact sciences and indispensable for unified and objective cognitive studies. Ten formalin-fixed human cadavers served for the studies. The preparations were prepared by the modified method of anatomical procedure. Linear and angular measurements of temporomandibular joint and masseter muscles were carried out with the use of the three-dimensional Cartesian system of OXYZ coordinates in relation to frontal, sagittal and horizontal planes. The physiological cross-sections of the masseter, temporal, lateral and medial pterygoid muscles were also determined. The collected data make it possible to develop a mathematical three-dimensioned model of the osseo-articulo-muscular system of the mastication organ.


Latent thyroid carcinoma in the material of the Institute of Oncology in Gliwice (Poland). The authors examined microscopically 110 randomly taken thyroid in serial sections. In 10 cases latent papillary thyroid carcinomas were found. The age and sex were without any influence on the incidence. Latent thyroid carcinomas incidence in this material was similar to that reported from the U.S.A., and Canada, but lower than in Japan and among Japanese population of the Hawaian Islands.


The parameters of cellular proliferation in acute (immature-cell) leukemias. In 20 patients with acute leukemia the 3H-thymidine labelling index and the mitotic index were determined in the population of leukemic blast cells and the degree of bone marrow infiltration was evaluated. Determinations were carried out in acute stage of the disease and in 4 cases additionally during improvement or remission and then during subsequent exacerbations. In the acute phase of the disease both indices were low in comparison with the indices obtained in normal granulocytopoiesis. In acute leukemia in 15 cases the bone marrow contained high or moderate number of cells, while in 5 remaining cases it was low. The 3H-thymidine labelling index and the mitotic index were lower in acute period of the disease in patients with bone marrow infiltration as compared with patients with hypoplastic bone marrow. During improvement both proliferation indices increased.


Effect of aggregated human gamma-globin on leukocyte migration-inhibitory test in patients with rheumatoid arthritis. In 69 patients with rheumathoid arthritis (RA): 49 sero-positive and 20 sero-negative for rheumatoid factor and in a group of 20 healthy blood donors the leukocyte migration-inhibitory test was carried out. Heat-aggregated gamma-globulin was used in the culture medium as antigen. Both groups of patients exhibited a significant inhibition of migration, as compared with the group of healthy subjects. However, a significant difference in the mean value of the migration indices between the sero-positive and sero-negative individuals was observed.


Role of disturbed metabolism of fat tissue cells in the pathogenesis of the diabetogenic effect of obesity in humans. It is supposed that the excess of fatty tissue exerts a diabetogenic effect. In obese subjects changes in the reactivity of the enlarged adipocytes to insulin might play a significant role in the pathogenesis of diabetes. Using the method of Rodbell in our own modification the responses of isolated adipocytes obtained from 10 lean and 15 obese subjects to a) insulin, b) theophylline and c) insulin and theophylline jointly (metabolism of glucose and glycerol) were determined. The dose-effect relationship curve was plotted against the effect of glucose utilization caused by increasing concentrations of insulin in the suspensions of adipocytes of lean and obese subjects. In adipocyte donors insulin sensitivity was also determined by Himsworth's test. It was found that adipocytes of 15 subjects with hyperthrophic obesity showed a significantly decreased effect of insulin regulating glucose utilization and glycerol release. They maintained a normal pattern of response to theophylline. The curve of relationship between insulin concentration in the incubation medium and its effect on glucose utilization by the adipocytes was changed. The adipocytes of these donors were relatively refractory to insulin in vivo as determined by the test of Himsworth. The cause of these disturbances may lie in the changed function of insulin receptors in the enlarged adipocytes. The observed disturbances may play a significant role in the pathophysiology of the diabetogenic effect of obesity.


Clinical and hemodynamic assessment of cardiomyopathy. A group of 43 patients with cardiomyopathy was assessed clinically and hemodynamically. Disturbances in the electric myocardial activity and hemodynamic changes concerning the duration of the different phases of left ventricular systole were observed. Despite the presence of these changes, in the early atypical forms no impairment of the myocardial contractility was found. In the advanced forms (hypertrophic cardiomyopathy and congestive cardiomyopathy) the myocardial contractility was impaired, this contributing to the development of heart failure manifestations.


Analgesic nephropathy--a continuing problem. Analgesic-induced renal disease occurs more commonly than is recognized; unless specific inquiry is made, it is seldom diagnosed. Patients with chronic pain, especially those with hypertension, urinary tract infection, or renal insufficiency, should be suspect. The condition is preventable and even in the late stages is manageable if recognized. Analgesic abuse may result in end-stage renal disease; it accounts for a significant percentage of patients entering chronic hemodialysis or renal transplant programs.


The treatment of connective tissue diseases with antilymphocyte globulin. Immunosupprissive therapy was carried out using steroids, azathiprine and antilymphocyte globulin in one patient with systemic lupus erythematosus (SLE)  and four with dermatomyositis. Response to treatment was based upon clinical evaluation. The patients with SLE and two of the four with dermatomyositis responded favourably to the regime. It is suggested that this preliminary study is sufficiently promising to warrant the creation of a rigidly controlled investigation.


The effect of intensive immunosuppression on the in vitro activity of lymphocytes from multiple sclerosis patients. During a small clinical trial of intensive immunosuppression in multipel sclerosis (MS, 14 patients) the changes of in vitro lymphocyte responses to mitogens were followed. A variable depression of the normal responses to phytohaemagglutinin (PHA), Concanavalin A (Con A) and pokeweed mitogen (PWM) was seen in the patinets during the inital week of treatment with prednisone (150 mg/day tapered to 20 mg/day by day 7), and azathiprine (3 mg/kg daily). They were further depressed during antilymphocyte globulin therapy (ALG 500 mg/day on weekdays, weeks 2-5 of treatment). These responses returned rapidly to the lower normal range after the three weeks of ALG despite the continued prednisone and azathipprine therapy. A complex effect of immunosuppression on lymphocyte subpopulations was suggested by three findings. Firstly, in contrast with the reduction in response to plant mitogens no lowering of the response to allogeneic lymphoid cell line cells (LCL) was seen during the first week of treatment. Secondly, some patinets, particularly those expressing the HLA-7 antigen, had a low pre-treatment response to LCL which showed an improvement during treatment and which was maintained through the first week of ALG treatment. Thirdly, single inviduals sometimes showed different degrees of suppression of different responses during and after ALG treatment. Occasionally, responses showed some recovery even during the ALG treatment, suggesting that higher ALG doses were required in these patients. The results suggest that the action of immunosuppressive drugs on lymphocyte activity differs between individuals due to the heterogeneity of the lymphocyte subpopulations present.


The treatment of autoimmune blood diseases with antilymphocyte globulin. Twelve patients with immunologically mediated diseases have been treated with ALG, alone or in combination. Tolerance was good. Indvidual case reports are discussed. Emphasis is placed on the duration of ALG-induced remissions, as well as on its favourable effect in humorally mediated autoimmune diseases. The absence of notable side effects, especially with the more recent and refined preparations, is stressed.


Antilymphocyte antiserum effect on incidence of spontaneous malignancy. I. Long term treated mice. Long term potent immunosuppression was administered to normal 7-week-old BALB/c mice without exogenous neoplastic induction. Antilymphocyte antiserum was given biweekly for 8 weeks and azathioprine daily for 347 days. Significant immunodepression was demonstrated after 348 days of theraphy. After 2 years, all animals were sacrificed and examined for neoplasia. There was no evidence that such immunodepression increased the incidence of spontaneous malignancy. The study suggests that immunosuppressive therapy is not inately oncogenic.


The validity and significance of hormone levels in gynaecology. The development of radioimmunoassay of steroid hormones in peripheral venous blood has led to a more direct index of gonadal production and concurrent monitoring of glandular function. While serial analyses of total urinary oestrogens can be correlated with luteinizing hormone (LH) excretion in treating amenorrhoea, radioimmunoassay of plasma oestrogens measures acute changes in ovarian responsiveness. For studies of androgens radioimmunoassay of plasma testosterone is the method of choice, but fractionation of urinary 17-oxosteroids may be useful and can be extended to include the pregnane series and corticoids. Urinary pregnanediol, however, may not reflect the endogenous secretion of progesterone and should be reserved for assessing induction of ovulation. Luteal function can be evaluated rapidly and specifically by radioimmunoassay of plasma progesterone. Although radioimmunoassay offers a new approach to ovarian physiology, its systematic application to disease is still in the future.


Steroids after birth-puberty.

Steroids after birth-puberty. The production of gonadal steroids in the perinatal period may have an important influence on the later development of the hypothalamo-pituitary-gonadal axis. In the prepubertal period there is an increased output of gonadal steroids and of androgens from the adrenal cortex, with a further increase as the secondary sex characters develop. The precise role of individual steroids in the maturation of the reproductive system and of the neuroendocrine regulating mechanisms will probably remain unclear unless conclusions are based on longitudinal studies in which hormone production is related precisely to different parameters of sexual development and somatic growth.


Anti-inflammatory action of corticosteroids. The anti-inflammatory action of corticosteroids is complex. At a cellular level, they cause redistribution of granulocytes, resulting in increased circulating granulocytes and reduced tissue pools. They also cause lymphopenia. The significance of these phenomena in relation to the anti-inflammatory activity of steroids is unknown. The most obvious pharmacological effects of corticosteroids are seen on blood vessels. They cause adrenergically mediated vasoconstriction and non-competitive antagonism of vasodilation due to prostaglanin E and bradykinin. Prostaglandin formation is inhibited by corticosteroids but whether this is due to an effect on enzymic synthesis or release is uncertain. Corticosteroids stabilize the lysosomal membrane preventing release of lysosomal enzymes in vitro but the significance of this in vivo is debatable.


The possibility of dietary protective factors in duodenal ulcer. II. An investigation into the effect of pre-feeding with different diets and of instillation of foodstuffs into the stomach on the incidence of ulcers in pylorus-ligated rats. Rats pre-fed on a South Indian diet show a much higher incidence of ulceration of the stomach after pyloric ligation than those on a North Indian diet. Removing vegetables and pulses from the North Indian diet deprives it of its protective effect. Staple foodstuffs with a high buffer content (unmilled rice, unrefined wheat and a millet [ragi]) placed in the stomach after pyloric ligation are also protective, but those with a low buffer content (milled rice, tapioca, sorghum and maize) are not protective. Refined wheat, however, gives protection despite its low buffer content. The significance of these findings in relationship to the distribution of duodenal ulcer in India and Africa is discussed.


Current problems in the measurement of gastrin release. A reproducible measure of physiological gastrin release. The interpretation of gastrin release is confused because of variation in the technique of the radioimmuoassay of gastrin, the lack of a standard stimulus for the release of gastrin and diversity in the method used to express the results. These problems have been analysed (a) by examining the cross-reactivity of three gastrin antisera and using each of the antisera to measure basal gastrin levels in fasting normal subjects, duodenal ulcer and post-vagotomy patients; (b) by determining a satisfactory stimulus for gastrin release in normal subjects; (c) by examining the results to determine the best method of presenting the data. The different a ntisera used were found to give different levels of plasma gastrin in the same sample of plasma. This was not related to the cross reactivity of the antisera. An English breakfast was found to be the most satisfactory stimulus for the release of gastrin. The expression of the results of such a stimulus of gastrin release was affected least by assay variation when the incremental integrated gastrin response was used. It is concluded that the incremental integrated gastrin response to an English breakfast is a satisfactory method for exploring variations in gastrin release.


The interrelationship between hypocomplementaemia, partial lipodystrophy and mesangiocapillary glomerulonephritis. A further case of mesangiocapillary glomth reduced serum C3 levels is reported. The father of the patient was also found to have C3 deficiency. This lends support to the hypothesis that C3 deficiency may be the primary disorder relating these conditions and there is evidence in this case that the deficiency may be genetically determined. It is suggested that the complement state of close relatives, particularly younger siblings, of patients with these disorders should be investigated and, if abnormalities are found, they should be followed-up in order to elucidate the role of hypocomplementaemia in these uncommon disorders.


Phaeochromocytoma with myocarditis managed with alpha-methyl-p-tyrosine. A case of bilateral phaeochromocytoma with catecholamine-induced myocarditis is described. The two operations needed allowed comparison of the use of alpha-methyl-p-tyrosine alone and in conjunction with adrenergic blocks in the management of the patient. The combination of both drugs was particularly successful in the relief of symptoms and reduction of catecholamine metabolism as monitored by 4-hydroxy-3-methoxymandelic acid (HMMA) excretion. As myocarditis is a potentially fatal complication, further investigation of the combined use of alpha-methyl-p-tyrosine and adrenergic blocking drugs is suggested in the pre-operative management of patients with phaeochromocytoma.


Caroli's disease with intrahepatic gall-stones and salmonella infection. At operation for small bowel intussusception, a 26-year-old man was found to have an enlarged liver and spleen. Subsequent investigations suggested bile passage infection associated with numerous intrahepatic gall-stones but symptomatic cholangitis did not present until 5 months later. Retrograde cholangiography showed cavernous ectasia of the bile ducts which contained gall-stones.


Systemic mastocytosis--central nervous system features and lymphographic demonstration of lymph node involvement. Systemic mastocytosis is uncommon. A patient is reported with many of the characteristic lesions of this disorder but with two features not before described--the occurrence of mixed pyramidal and neuromyopathic neurological signs and the demonstration of lymph nodes with abnormal architecture by lymphography.


Metastatic endobronchial hypernephroma nine-and-a-half years after nephrectomy. A patient with an endobronchial nodular lesion, proved to be clear cell carcinoma (hypernephroma) by fibrebronchoscopic biopsy, is described in which the primary tumour had been resected 9 1/2 years before the onset of the present illness. At post-mortem the remaining kidney was found to be normal.


Nephrotic syndrome, sickle-cell disease and pregnancy. A young West Indian with sickle-cell disease and the nephrotic syndrome in the last trimester of pregnancy was treated after admission to hospital with folic acid and blood transfusion. Labour was induced by intravenous prostaglandin E2 but the fetus died. The nephrotic syndrome resolved but the proteinuria, the cause of which it is suggested is a true sickle-cell nephropathy, persisted.


Evaluation of heterogeneity in hemoglobins of Gallus domesticus and coturnix quail using disc-gel electrophoresis. The heterogeneity in the hemoglobins of Gallus domesticus and Coturnix quail (Japonicum) was studied. Comparisons between a mutant hemoglobin type, the normal hemoglobin type and a heterozygous type were made in both species using disc-gel electrophoresis. The same heterogeneity of the primary hemoglobins as had previously been observed in these populations of chickens and quail by cellulose acetate electrophoresis was also observed using disc-gel electrophoresis. The same major hemoglobin component was observed in all the hemoglobin types, but the electrophoretic mibility of the minor component was different. In addition to the major and minor components which accounted for 93% of the total hemoglobin, four trace components were observed. One of these trace components (T) had previously been shown by cellulose acetate electrophoresis to be associated with the mutant minor hemoglobin. The other three trace components were in close proximity, but migrated anodally to the major, normal minor and mutant minor components. Their presence was not consistent and their concentration was usually at the lower limit of resolution. No differences could be detected between the electrophoretic mobility of the mutant hemoglobin of chickens and quails, suggesting that parallel mutations have occurred in the two species.


Grwoth response of Muscovy ducklings to thyroxine (iodinated casein). Iodinated casein at 220 mg. kg. of feed was fed to Muscovy ducklings to determine its effect on sexually differentiated growth. Nine male and eight female treated ducklings averaged 2,135 and 1,520 g. at eight weeks as compared to 2,315 and 1,588 g. for the untreated nine males and eight females. Feed conversion was 2.99 for treated and 3.60 for untreated ducklings.


The effect of feeding various levels of dietary glycine in a pre-experimental diet to one-day old chicks on their subsequent glycine plus serine requirement. Two experiments were conducted to determine the effect of feeding chicks 1=9 days of age various levels of clycine in corn-casein and corn-soybean-corn gluten meal diets upon the subsequent glycine plus serine requirement of chicks during the remaining 10-21 day feeding period. Chicks in Experiment 1 fed corn-casein basal diets with 1.3% additional dietary glycine (2.15% total glycine plus serine) gained significantly (P less than or equal to 0.01) more weight during an initial 9-day feeding period than chicks fed basal diets with 0.3% and 0.9% supplemental glycine. Chicks fed the 2.15% glycine plus serine diets during the first 9 days posthatching and then fed basal diets containing 0.3% supplemental glycine (1.15% total glycine plus serine) for a 10-21 day feeding period gained weight equivalent to chicks fed 2.15% glycine plus serine diets for the entire 21-day period. The corn-casein basal diet contained 21.0% protein and 0.85% glycine plus serine. Chicks in Experiment 2 fed corn-soybean-corn gluten meal diets containing 1.8% glycine plus serine did not respond to glycine supplementation druing the initial 9-day feeding period or the 10-23 day feeding period. The results suggest feeding optimum levels of glycine to chicks during the first nine days after hatching decreases the requirement of glycine and serine during subsequent feeding periods.


Dietary restrictions during adolescence and subsequent reproductive performance of turkey breeder males. A study was conducted to investigate the effect of early dietary restriction of protein and/or energy on the subsequent reproductive ability of the tom. One hundred 8-week old Large White turkey toms (April hatch) were placed at random into four floor pens of 25 each and fed the following diets: group 1, 20.6% protein with 2,4 Mcal. M.E./kg. diet 8-12 weeks, 15.3% protein with 2.68 Mcal. 12-16 weeks, 10.4% protein with 2.85 Mcal. 16-24 weeks; group 2, 20.3% protein with 2.9 Mcal. 8-12 weeks, 14.8% protein with 2.98 Mcal. 12-16 weeks, 9.8% protein with 3.27 Mcal. 16-24 weeks; group 3, 22% protein with 2.4 Mcal. 8-12 weeks, 18.9% protein with 2.57 Mcal. 12-16 weeks, 16.3% protein with 2.81 Mcal. 16-24 weeks; group 4 (control), 22% protein with 2.91 Mcal. 8-12 weeks, 19% protein with 3.03 Mcal. 12-16 weeks, 15.9% protein with 3.17 Mcal. 16-24 weeks. Beginning with the 25th weeks all toms received a 14.3% protein diet with 2.8 Mcal. The restriction of energy significantly reduced the percentage of males producing semen, and the restriction of energy significantly reduced the percentage of males producing semen, and the restriction of energy and both protein and energy significantly depressed semen volume per male in comparison with the average produced by the control males. Other reproductive traits were not affected by early dietary controls.


Leucine, isoleucine and valine interactions in turkey poults. In four experiments, the interactions of leucine, isoleucine and valine in turkey poults were studied. The additon of 1.50% excess leucine to a 22% protein starter diet, marginal in isoleucine and valine, depressed growth. This growth depression was corrected by the addition of valine and isoleucine. The addition of the excess leucine caused a decrease in plasma valine and isileucine concentrations in experiments 3 and 4, and plasma valine concentration in experiment 1. The addition of valine caused a marked linear increase in plasma valine with little or no effect on plasma isoleucine. The addition of isoleucine to the diet caused an increase in plasma isoleucine. Plasma valine, however, was decreased by the addition of isoleucine to a high-leucine diet. It is concluded that interactions exist in turkey poults between leucine-valine, leucine-isoleucine and isoleucine-valine and that the growth reduction caused by added leucine can be partly alleviated by addition of valine or by valine plus isoleucine, but not be isoleucine alone.


Lysozyme in hen blood serum.

Lysozyme in hen blood serum. Hen blood serum (White Leghorn) possessed the lytic action against Micrococcus lysodeikticus which was less than one-thousandth of egg white obtained from hens of the same species, suggesting that lysozyme was present. The filter-sterilized hen blood serum also inhibited the growth of M. lysodeikticus in broth culture. The isolated lysozyme, purified by column chromatography and gel filtration, proved to be a basic protein with a low molecular weight (about 15,000), active against M. lysodeikticus, and more stable at acidic pH values than at alkaline pH values when heated. In amino acid composition, the isolated serum lysozyme had slightly higher proline and lower aspartic acid content than hen egg white lysozyme. The blood serum lysozyme was less heat stable at various pH values (4.5 to 8.4) than egg white lysozyme.


Effect of heat stress on the potassium requirement of the hen. Four diets were formulated with potassium levels ranging from 0.4 to 1.0% and each diet was fed to laying hens at temperatures of 26.7%, 32.2, and 37.8 degrees C. Analyses for potassium in the plasma, egg, and excreta and potassium retained were used to determine the effects of heat stress on the potassium balance of laying hens. Urine potassium data from a surgically-altered hen provided insight into urinary excretion of potassium under conditions of heat stress. Regression analyses of the data obtained indicated that temperature significantly influenced plasma potassium concentrations, while egg and excreta potassium concentrations and the amount of potassium retained were not significantly influenced by temperature. The data suggest that 0.4% potassium in the diet is adequate for meeting potassium requirements under conditions of heat stress. However, 0.6% dietary potassium may be a more suitable level for assurance against a potassium imbalance at elevated temperatures. One percent dietary potassium appears to be an excessive level.


Partial purification of the ovarian oviposition-inducing factor and estimation of its chemical nature. The ovarian ruptured follicles larger than 0.2 g. of wet weight were collected from laying hens and an ovarian oviposition-inducing factor (OOIF) was extracted from the follicles. The extract was chromatographed on a column of Sephadex G-75. Material from the active region for inducing premature oviposition was subjected to thioglycollate or trypsin treatment. It was found that oxytocic activity of this material was completely destroyed by thioglycollate buy not by trypsin, when tested on the contractility of the isolated hen's uterus. The OOIF was found to be diffusible through a cellophane bag. It is suggested that the OOIF may be a relatively small molecule have a disulfied bridge and it seems to be a hormone or hormone-like substance resembling the posterior pituitary hormones in the chemical nature.


A comparison of various routes of Newcastle Disease vaccination at one day of age. Groups of 100 one-day-old chicks with moderate levels of maternal antibody were vaccinated with the B1 strain of Newcastle disease virus subcutaneously, intratracheally, intraocularly or by aerosol or were left unvaccinated. Certain of these groups were revaccinated at 9 or 9 and 28 days or at 14 or 14 and 28 days with the LaSota strain of Newcastle disease virus. Vaccination by all routes at one day of age resulted in an antibody response and increased resistance to challenge. The most effective route was by aerosol followed by the intraocular method. Intratracheal administration or subcutaneous inoculation led to a marginal response. Aerosol administration at 9 or 9 and 28 days or at 14 or 14 and 28 days gave adequate antibody responses and resistance to challenge.


Studies on avian erythrocyte metabolism. 5. Relationship between the major phosphorylated metabolic intermediates and while blood oxygen affinity in embryols and poults of the domestic turkey (Meleagris gallopavo). The changes in organic phosphates of turkey erythrocytes (RBC) have been determined in relation to the changes in oxygen affinity of whole blood during growth of the embryo and poult. On a molar basis, 2,3 diphosphoglyceric acid (2,3-DPG) is the predominant organic phosphate of erythrocytes from turkey embryos during the last week of incubation. However, on the basis of relative % phosphate, 2,3-DPG is the major organic phosphate of the erythrocytes from turkey embryos on day 23 and 25 of incubation only. With the exception of day 23 and 25 incubation, adenosine triphosphate (ATP) represents the major organic phosphate of the erythrocytes of the turkey embryo and poult during the last week of embryonic development and through the first 29 days after hatching. The whole blood P50 during the last week of incubation and the first 8 days after hatching correlates best with the amount of ATP in the erythrocytes. The effects of inositol pentaphosphate on P50 of the whole blood is much more gradual and appears to become of major influence after 2-3 weeks post-hatching.


A study of Salmonella contamination in broiler flocks. The study was designed to follow the Salmonella infection rate in a poultry flock during the growth period until processing at 8-9 weeks. The infection rate was as high as 37.5% initially and dropped significantly by the 3rd to the 5th weeks. There was no difference between various types of litter and the species of Salmonella isolated. Little correlation was found between Salmonella species in feed ingredients and those identified from the flocks.


Influence of arginine vasotocin on a genetically determined excessive appetite for water in chickens. Females from two inbred strains of chickens, one normal and one having hereditary diabetes insipidus, were treated with five levels (2.5, 1.0, 0.5, 0.25, and 0.1 units) of Schwartz-Mann (Grade A, 100 units/mg.) arginine vasotocin via ic. injections at 2-hr. intervals. Paired sibs, one normal and one having diabetes insipidus (di), from an F2 population were used in one experiment to compare responses at the level of 2.5 units. Only di females were used for the remaining four levels of treatment. Water intake was determined every two hours and calculated as a percentage of water (ml.)/body wt. Results show that hereditary diabetes insipidus in chickens is arginine vasotocin-sensitive at the 0.5 unit and higher levels. The genetic defect appears to affect the quantity of antidiuretic hormone produced.


Long term studies comparing the efficacy of cell-free versus cell-associated HVT vaccines against Marek's disease. In long-term field trials lasting approximately 12 months attempts were made to determine whether the cell-free HVT vaccine was as effective in reducing MD losses as the cell-associated HVT vaccine. Four trials were conducted in which flocks were vaccinated with either the cell-free or the cell-associated HVT vaccine. The cell-free HVT vaccine was as safe and effective in preventing MD as the cell-associated HVT vaccine. Although there were some losses due to MD in all flocks, there were also losses due to lymphoid leukosis, multiple hemangioma, liposarcoma or coccidiosis. Egg production was essentially the same in flocks vaccinated with the cell-free HVT vaccine as in those vaccinated with the cell-associated HVT vaccine.


Influence of Fusarium-infected corn and F-2 on laying hens. Diets containing either Fusarium-infected corn supplying 25 and 100 p.p.m. of F-2 (zearalenone) or purified F-2 at these levels did not adversely influence the reproductive performance of laying hens. In trial 1, no deleterious effects were observed for 20- and 36-week body weights, age at first egg, egg weight, albumen height, shell deformation, fertility or hatchability when Fusarium-infected corn was fed to 20-week-old pullets for 28 days. Percent hen-day egg production of birds fed Fusarium-infected corn supplying 25 and 100 p.p.m. of F-2 was superior (P less than or equal to 0.5) to egg production of nontreated controls. In trial 2, three replications of ten adult Leghorn hens were evaluated under five dietary treatments: (1) 16.7% protein basal; (2) basal plus Fusarium-infected corn (25 p.p.m. of F-2); (3) basal plus 25 p.p.m. of (purified) F-2; (4) basal plus Fusarium-infected corn (100 p.p.m. of F-2); (5) basal plus 100 p.p.m. of (purified) F-2. Difference between dietary treatments for 14-day pre-treatment, treatment and post-treatment periods were nonsignificant for 42- and 44-week body weights, egg production, egg weights, fertility and hatchability. Body weights of chicks from hens fed F-2 diets were not significantly different from those of chicks from hens fed the basal diet.


Histochemical observations on the seasonal fluctuations in the follicular atresia and interstitial gland tissue in the house sparrow ovary. A histochemical study has been made of seasonal fluctuations in the follicular atresia and interstitial gland tissue with the ovarian cycles of the house sparrow (Passer domesticus). The atresia of primordial oocytes forms the predominant feature of the quiescent-winter ovary. The building up of interstitial gland tissue of thecal origin. which precedes the breeding activity, is closely related to the atresia of previtellogenic follicles of variable sizes. Interstitial gland cells which accumulate lipid droplets consisting of cholesterol, and/or its esters, triglycerides and some phospholipids, are the steroid secretors. The possible functional significance of different-sized follicular atresia in relation to follicular growth and breeding activity is discussed.


Cytogenetic and phenotypic effects of a chromosomal rearrangement involving the Z-chromosome and micro-chromosome in the chicken. Measurements demonstrated that the Z-chromosome was truly metacentric. Forty-six percent of one arm of a female's Z-chromosome had been translocated to a microchromosome (Z-micro) by irradiation of semen. The bread was 23 crossover units distal to the late feathering (K) locus. The barring (B) locus on the non-broken arm assorted almost independently of the Z-micro segment. Semen from eight sons of this Z-micro female was used to inseminate 98 dwarf (dw) broiler-type females. From karyotypes of 147 male and 149 female progeny, we identified 69 males heterozygous and 79 females hemizygous for Z-micro. Body weight of 43 males heterozygous for Z-micro was significantly greater than that of 45 normal Z paternal half-brothers at all ages from 2 to 24 weeks. In contrast, body weight of 57 Z-micro females compared with their 56 normal Z paternal half-sisters was depressed significantly at 2, 4, and 6 weeks but not significantly at 8, 12, 16, and 24 weeks of age. Age at first egg was retarded 8 days and egg production over a 153 day test period was reduced 19.6%, primarily due to a reduction of egg laying sequence from 2.7 to 2.1 days in the Z-micro females.


Metabolic responses of White Pekin ducks to ambient temperature. Metabolic responses of unrestrained White Pekin ducks, Anas platyrhynchos, were determined at ambient temperatures of -5 degrees, 1 degrees, 10 degrees, 20 degrees, and 30 degrees C. Temperatures from the pectoralis and the subcutaneous back were monitored. Unrestrained ducks exhibited two metabolic states, a high response (540 cc. O2/kg/hr. at 20 degrees C.) and a low response (427 cc. O2/kg./hr. at 20 degrees C). Pectoralis muscle temperature did not vary greatly over the range of ambient temperatures tested and was considered to be a good representative of body temperature.


Accudosetm aerosol--an effective house fly (Diptera; Muscidae: Musca domestica L.) adulticide control system for cage type poultry houses. An automatic piped aerosol system (Accudose TM) using 0.7% synergized pyrethrin insecticide, was tested at a typical narrow cage poultry farm for the control of adult house flies, Musca domestica L. A similar narrow cage poultry farm was used as a control with all house fly control measures left up to the cooperating poultryman. AccudoseTM was compared with other house fly control methods at other similar farms which included three types of man-portable ultra volume (ULV) generators and an integrated (biological-chemical) program. Results of the five month test demonstrated that the AccudoseTM system suppressed house fly populations better than other control methods.


Liver lipid accumulation and performance of hens as affected by cage density and initial body weight. Hens housed in individual cages (25  46 cm.) laid significantly more eggs, consumed significantly more feed and had significantly larger livers with a higher lipid content than hens housed three to a cage in two experiments. Body weight gain was significantly higher for individually housed birds in experiment 1, but not in experiment 2. No difference was observed in mean egg weight or kg. feed per dozen eggs. In experiment 2 hens housed two to a cage laid slightly fewer eggs and accumulated slightly less liver fat but the differences were not significantly different fromthose housed individually. Hens socially dominant in three bird pens had higher liver fat accumulation than hens lower on the peck order but liver fat accumulation for the dominant hens still averaged less than hens housed either two or one per cage. Comparison of two strains in experiment 1 revealed a significant difference in rate of egg production and feed efficiency but no difference in liver fat accumulation. Pullets placed in four body weight classes prior to the start of the experiment did not differ significantly in liver fat accumulation per unit of body weight or percentof dry matter of the liver at the end of the experiment. Rate of egg production and feed efficiency were also not significantly different among the body weight classes.


Toxicity and tissue residues from diphenylhydantoin fed to chickens. Within three experiments with broiler-type crossbred chickens, diphenlyhydantoin (DPH) was administered via the diet at levels ranging from 10 to 2500 mg./kg. Feed intake and body weight gain were adversely affected by levels of 100 mg./kg. or higher. A hemorrhagic syndrome occurred in 16 percent of the chickens fed 100-1000 mg./kg. and 33 percent of those fed 2500 mg./kg. Twenty percent of the chickens fed 1000 mg./kg. were anemic; none of those receiving 250 mg./kg. were anemic. Other toxic manifestations included neurological side effects of a lethargic stupor, excitability to a disturbance, or a characteristic condition referred to as "head under." Pair-feeding experiments revealed thatlowered feed intake was not responsible for any of the clinical signs. All of the toxicity signs were reversible in 3 to 4 weeks following withdrawal of DPH. Muscle samples contained 2.5 to 11.4 mcg. DPH/g. at drug levels of 1000 and 2500 mg. DPH/kg. diet. Dose-response curves revealed that plasma, adipose, kidney and liver had levels which were, respectively, 1.05, 1.51, 2.19, and 2.85 times those of muscle.


Effect of high amounts of dietary zinc and age upon tissue zinc in young chicks. Weight gains of younger broiler chicks were not reduced by up to 2400 p.p.m. added zinc fed to four weeks of age. Tissue zinc was not changed significantly by 600 or 1200 p.p.m. supplemental dietary zinc, but at 2400 p.p.m. added zinc, blood, kidney, and liver zinc were significantly elevated (P less than 0.05). In the heart, zinc was not affected by the added dietary zinc but increased over time with all diets. Unlike the calf and weanling pig, the metabolism of zinc in the young chick does not appear to change appreciably with increasing maturity. Thus, the homeostatic control mechanisms of the young chick are effective for up to 1200 p.p.m. dietary zinc, but are partially overcome by 2400 p.p.m. added dietary zinc.


Manganese levels and the morphology of the epiphyseal plate in broilers with slipped tendons. Manganese deficiency in chickens results in perosis and a higher incidence of slipped tendon. Perosis is associated with a disorganization of the epiphyseal growth plates and changes in the chemical composition of the cartilage matrix. Male broilers with slipped tendons were selected from a commercial broiler farm over a 9 week growing period. Their growth rates, epiphyseal cartilage histology and tissue manganese concentrations were examined and compared with (a) normal broilers from the same farm, and (b) similar broilers raised on control and manganese deficient diets. Field broilers with slipped tendons showed no evidence of abnormality in the histological appearance of the proximal tibial growth plate at any of the ages examined. The manganese content of liver and epiphyseal cartilage from field broilers showing slipped tendon was comparable with that present in tissue from normal broilers wither from the field or raised on a chemically defined diey supplemented with managese. The slightly retared growth rate seen in the field broilers was attributed to feeding problems associated with the lameness condition. These results provide conclusive evidence that slipped tendon in field broilers is not dur to a manganese deficiency in the tissues, nor does it appear to be associated with abnormal proliferation of the chondrocytes in the epiphyseal plate.


A simplified biochemical system to screen Salmonella isolates from poultry for serotyping. The 24 most frequently isolated paratyphoids from poultry, along with Salmonella gallinarum and Salmonella pullorum, plus strains of Arizona, Citrobacter, Edwardsiella, Escherichia, Klebsiella, Prteus, Pseudomonas, Serratia and Shigella were inoculated into triple sugar iron (TSI) and lysine iron (LI) slants and into six fermentation broths which were numbered: 1 (dextrose), 2 (lactose), 3 (sucrose), 4 (mannitol), 5 (maltose), 6 (dulcitol). All the salmonella cultures (except S. pullorum) gave a 1, 4, 5, 6 code which means they produced acid, and in most cases gas, in dextrose, mannitol, maltose and dulcitol, but no acid or gas in lactose and sucrose. S. pullorum gave a 1, 4 code. All non-salmonella cultures gave a fermentation pattern different from the 1, 4, 5, 6 pattern of paratyphoids and S. gallinarum. Therefore, this six sugar system can be successfully used in the selection of suspect salmonella cultures for specific typing. Results from a miniaturized system (Minitek) were the same as those from the standard tube method for the carbohydrate fermentation tests for all cultures tested.


A simplified technique for obtaining purified peripheral lymphocytes from chickens. A simplified technique for isolating purified avian peripheral lymphocytes using a Ficoll-diatrizoate density is described. The procedure requires only 2 ml. of blood, can be completed in 5 min., and is  suitable for the repeated testing of relatively large numbers of birds.


Effects of dietary citrinin and aflatoxin on leg problems of broilers growtn on wire floors. Dietary levels of citrinin of 62.5, 125 and 500 mug./g. diet or aflatoxin at a dietary level of 2.5 mug./g. of diet fed from 1-21 days of age did not inccrease the incidence of leg problems compared to control groups receiving no dietary nycotoxins. These data indicate that dietary contamination with these toxins does not account for the incidence of occasionally observed leg probelms in younger broiler chicks.


A feed additive to control flies in poultry manure. To determine methoprene's effect on fly emergence from poultry manure, 18,000 8-wk.-old caged pullets were fed continuously 10 gm. of methoprene per ton of feed 42 days. Hatch mates (13,500) in an adjacent, similarly constructed and equipped house were fed untrated feed. Data on fly emergence were recorded twice a week after counting average numbers of flies in fly traps, in buckets of manure, and on fly strips and spot cards. Differences between average counts in the treated and control groups were significant by each of the four data-gathering methods. The methoprene reduced fly emergence by 77.0% in manure buckets, 65.5% in fly traps, but increased fly emergence 19.2% on fly strips, and 92.2% on spot cards. We attribute the increases in fly counts by the latter two counting methods in the treated house to migration of flies from adjacent untreated houses.


Forensic problems in practice. VII.--Asphyxia. In summary, asphyxial deaths and deaths due to pressure on the neck form a considerable proportion of homicides in Britain, as well as many suicides and some accidents, as in infancy and in masochistic practices. Although the detailed examination of fatal asphyxia is the province of the coroner's pathologist, the general practitioner and the casualty officer are usually the first medical men to have the opportunity of detecting the external signs of such cases. Unfortunately, very many instances in the past have revealed that the clinician's examination of dead persons is so superficial that numerous cases of trauma to the neck (and even florid asphyxia) have gone unnoticed. A rapid examination of the skin, conjunctivae and neck takes but a few seconds and, if done routinely, would save subsequent embarrassment to the doctors concerned.


Seizures after the age of sixty.

Seizures after the age of sixty. Despite adequate diagnostic evaluation and prolonged follow-up, the causes of epilepsy in the elderly patient frequently remain unknown. Some of these may be of occult, degenerative or vascular central nervous system disease origin--entities which are often difficult to incriminate specifically. Unquestionably, in a few cases, seizures occur at the time of the initial cerebrovascular accident, although in some patients seizures may follow such an episode after a latent interval, particularly when cortical motor areas have been damaged by the initial vascular insult. It is important to differentiate this group from those with recurrent vascular episodes, in whom diagnostic studies and treatment may differ. Brain tumour is as prominent a cause of seizures in the seventh decade as it is in younger adults, but probably is of lower incidence thereafter. Focal, clinical and electroencephalographic findings and a progressive clinical course are particularly common in this group. A heterogeneous group of causes, a few of which are liable to affect the ageing patient, are responsible for some of the seizures in old age and should be searched for when the etiology is obscure. Diagnostic studies will vary with the individual problem and, whenever possible, should be restricted to non-invasive studies such as electroencephalography and computer-assisted tomography, after initial baseline clinical assessment and laboratory studies have been performed. Contrast procedures such as angiography are sometimes hazardous to patients with atherosclerosis and should be reserved for selected patients. Treatment follows conventional guidelines with some modifications due to the vulnerability of the ageing patient to degenerative disease and complications of immobilization.


Innocent heart murmurs in children. A survey of 119 patients. The clinical findings in 119 children with innocent heart murmurs have been reviewed. The majority were aged between 1 and 5 years, and it was in this age-group also that a maximum incidence of venous hum was recorded. There were no particular ECG or chest X-ray findings.


Foreign bodies in the larynx in infancy. Three cases of foreign bodies in the larynx are described. The mode of presentation, management, and the importance of early diagnosis to decrease the number and severity of complications are stressed.


[Hyperlipidemias and abnormalities in the electrophoresis of lipoproteins. Incidence in newborn infants]. We studied 300 newborns in whom total cholesterol, cholesterol of low density lipoproteins, triglycerides and lipoprotein electrophoresis were determined in umbilical cord blood. Pure hypercholesterolemia was found in 14 cases (4.7%), hypertriglyceridemia in 15 cases (5.0%) and increase in cholesterol and triglycerides in 4 cases (1.3%). Lipoprotein electrophoresis showed increase in betas in 23 cases (7.6%), prebetas in 24 (8.0%) and in both, betas and prebetas, in 6 cases (2.0%). Betas /alphas index was always found elevated when other electrophoretic fractions had some abnormality.


[An alcohol-soluble trypsin inhibitor from beans]. An ethanol soluble protein-trypsin inhibitor was isolated from kidney bean seeds. The inhibitor purification included gel chromatography of ethanol soluble proteins on Sephadex G-75 and affine chromatography on immobilized trypsin and chymotrypsin. The inhibitor suppressed activites of trypsin, chymotrypsin and in part trypsin-chymotrypsin activity of pronase but did not influence subtilizin. The inhibitor at a dose of 15-17.5 mug decreased by 50% the activity of 100 mug trypsin. By isoelectric focusing it was shown that the inhibitor isolated from kidney beans consisted of four isoinhibitors with pH of 4.3, 4.5, 4.7, and 4.9.


[Catalase inactivation during storage in solution and its stabilization by polysaccharides of microbial origin]. It has been experimentally shown that the rate of inactivation of pure catalase in solution does not obey the kinetics of the first order. The kinetics of denaturation taking into account partial stabilization of catalase due to the formation of intermolecular complexes of native molecules with denatured molecules has been derived. Using the equation, the rate of catalase inativation during storage in pure solutions has been calculated from the experimental data. Polysaccharides of microbial origin can also stabilize catalase. The kinetics of catalase inactivation in the system where the enzyme and polysaccharide forms a reversibly dissociating complex has been described.


Approaches to the study of acetylcholine turnover in brain. All of the methods presently available for estimating acetylcholine turnover have severe limitations and rest on unproven and sometimes fallacious assumptions. Taken individually, they may be misleading. To the extent that the assumptions are different for different approaches, agreement between several approaches may provide satisfactory evidence regarding normal turnover rates and changes which may be experimentally induced.


Effects of chronic morphinization on calcium membrane interaction. Tolerance to morphine proceeds with an adaptation in the nerve membrane causing an increase in endogenously bound calcium. Our data on calcium binding is compatible with Collier's hypothesis (10) for drug induced changes in number and/or types of receptors. The results demonstrating naloxone reversal of this membrane adaptation suggest that opiate antagonist action may be to maintain the opiate macromolecule portion of the membrane in a Ca++ associated homeostatic state. Lack of effects of Na+ on opiate treated SPM suggest that cellular adaptation to morphine proceeds in vivo through a change in membrane binding of calcium.


The effect of memory blocking antibiotics and their analogs on acetylcholinesterase. The ability of antibiotics to inhibit acetylcholinesterase was measured in homogenates of goldfish brain. Puromycin aminonucleoside was the most potent inhibitor followed by puromycin, cycloheximide and acetoxycycloheximide. Puromycin effectively impaired retention of active-avoidance learning in goldfish when injected either immediately before or after training, while puromycin aminonucleoside did not regardless of injection time. These results suggest that the known amnestic effects of puromycin, cycloheximide and acetoxycycloheximide are not a consequence of interference with acetylcholinesterase.


Effects of intracisternal GABA or glutamic acid upon behavioral activity in the rat. It has been postulated that GABA acts as an inhibitor of command neurons, the activity of which initiates behavior. A prediction of this hypothesis is that elevations in functional GABA levels in the brain will cause decreases in behavioral output. Accordingly, in this study rats were injected intracisternally with either saline or one micromole of GABA or its excitatory precursor, glutamic acid, and behavioral activity in a novel environment was recorded as it habituated over the course of the subsequent 4 hours. The activity of the animals that were injected with GABA was greatly decreased, while the activity of the animals that were injected with glutamic acid was apparently unaffected, as compared to animals given saline. These data provide support for the hypothesis that GABA functions as an inhibitory neurotransmitter for behavior-activating command neurons.


The effect of 6-aminodopamine on electrical self-stimulation in rats. The compound 6-aminodopamine is a powerful CNS catecholaminergic neurotoxin. Small dosages of 6-aminodopamine injected intraventricularly markedly depress electrical self-stimulation rates in rats. This 6-aminodopamine treatment produced whole brain lowering of norepinephrine to ca. 50% of normal while the dopamine content was unchanged. The possible use of 6-aminodopamine treatment to elucidate the relative roles of norepinephrine and dopamine pathways is discussed.


The drinking response of the chicken to peripheral and central administration of angiotensin II. Intravenous injection of Ang II (val5 angiotensin II amide) elicited an immediate drinking response in the domestic fowl which lasted at least 20 minutes. The minimal dosage needed was 300 mug. Intracranial injection of 10 mug Ang II through cannulas implanted in the anterior diencephalon caused a significant increase in water intake. The minimal intracranial dosage of Ang II which evoked drinking was 2.5 mug. Intracranial injection of isotonic KCl inhibited the drinking response induced by intravenously injected Ang II when administered simultaneously. This suggests that drinking caused by both intravenous or intracranial injection of Ang II is activated through identical brain regions. The positive drinking response of the chicken is repeated concecutive intracranial injections of Ang II declined from the first injection through the following ones.


Genetic differences in ethanol drinking of the rat following injection of 6-OHDA, 5,6-DHT or 5,7-DHT into the cerebral ventricles. The preference characteristics for ethanol of four different strains of rats were determined in a two-choice situation by offering water and ethanol in a concentration which was increased from 3 to 30% over a 12-day test sequence. Using stereotaxic procedures, 50 mug 5,6-dihydroxytryptamine (5,6-DHT), 200 mug 6-hydroxydopamine (6-OHDA) or 100 mug 5,7-dihydroxytryptamine (5,7-DHT) were then injected acutely into the lateral cerebral ventricle in a 20 mul volume. Rats of the Sprague-Dawley strain increased their ethanol preference following the lesioning of the serotonergic system by 5,6-DHT, whereas similar destruction of catecholaminergic neurons by 6-OHDA markedly suppressed ethanol intake; Long-Evans rats displayed a similar trend in ethanol drinking patterns. However, animals of the Holtzman strain manifested the increased drinking after 5,6-DHT, but showed no suppression of drinking following 6-OHDA. The preference of rats of the Wistar strain was unaffected by 5,6-DHT but attenuated by 6-OHDA. 5,7-DHT had little or no effect on ethanol consumption in any of these strains. These findings thus suggest that genetic factors are an important determinant in an animal's response to a drug that affects 5-HT or NE systems in the brain, particularly when ethanol selection is investigated.


Hypothermia induced by delta9-tetrahydrocannabinol in rats with electrolytic lesions of preoptic region. The preoptic region (POR) is a primary central site for thermoregulation. Bilateral lesions of POR disrupt thermoregulation, and in rats, produce a characteristic syndrome including hyperthermia. delta9-Tetrahydrocannabinol (delta9-THC), a potent hypothermic agent,  appears to mediate this effect via some central mechanism. The studies reported here suggest that delta9-THC induces hypothermia at a site other than POR. Male Sprague-Dawley rats were divided into 2 groups, one with subsequently confirmed bilateral POR lesions and a sham operated group. The lesioned animals developed hyperthemia (+2.1degrees +/- 0.1degreesC, p less than 0.01) within 2 hr after surgery when compared to the sham operated controls. delta9-THC was administered intraperitoneally (5 and 10 mg/kg). Rectal temperature was recorded at 30 min intervals for 2.5 hr. Both lesioned and nonlesioned rats exhibited hypothermia within 30 min of delta9-THC administration. The hypothermic response to 5 and 10 mg/kg delta9-THC in the lesioned animals was significantly greater (p less than 0.05) and showed a trend toward longer duration than the hypothermia induced in the sham operated controls. These data demonstrate that delta9-THC is able to induce a hypothermic response in rats whose body temperatures were elevated by POR ablation. Although delta9-THC does not appear to act primarily at POR to induce hypothermia, it is evident than an intact POR plays a role in modifying the duration and magnitude of delta9-THC induced hypothermia.


Impairment of Hebb-Williams maze performance following prolonged alcohol consumption in rats. Wistar rats consumed a liquid diet containing sustagen and ethanol for a period of 150 days. Control animals were pair-fed an identical diet except for isocaloric substitution of sucrose for ethanol. Forty-five days after the liquid diets were discontinued both groups were tested for performance on the Hebb-Williams maze. The experimental group made significantly more errors than the control group indicating that long-term ingestion of alcohol can lead to substantial impairment of problem-solving abilities in rats.


Effects of handling and etherization on incorporation of [3H] lysine into protein of mouse brain and liver. A standardized handling experience of 20 sec duration elevated the amount of radioactive lysine incorporated into brain total proteins during a 10 min labeling period begun either 10 min or 30 min after the handling. Etherization for 20 sec produced similar metabolic changes. Incorporation of [3H] lysine into liver proteins was minimally affected by handling, while slightly altered by etherization. The metabolic changes detected in the brain after handling did not appear to be side-effects of changes in blood-borne radioactivity. The results indicate that laboratory stresses often thought to be of minor importance can have large effects on ordinary assays of protein metabolism in brain.


Effect of two inhibitors of dopamine beta-hydroxylase on maturation of memory in mice. Bitemporal injections of puromycin that primarily affect the hippocampal-enthorhinal cortical areas suppress memory of maze-learning in mice for 3 days after training but are ineffective 6 or more days after training. At these later times, injections affecting widespread areas of the brain in addition to the hippocampal-entorhinal area are necessary for amnesia. These observations are interpreted to indicate that the locus of the memory trace has enlarged at 6 days to include other parts of the central nervous system in addition to the hippocampal-entorhinal area. To produce an imbalance of neurotransmitters and so to test their importance in enlargement of the memory trace's locus, we treated mice for 7 days after training with inhibitors of dopamine beta-hydroxylase. These mice, unlike untreated controls, developed amnesia after bitemporal injections of puromycin. In view of additional control experiments, we interpret these results to suggest that an imbalance of transmitters suppresses the normal enlargement of the locus of the memory trace.


Neurochemical and behavioral effects of diet related perinatal folic acid restriction. Some effects of restricting dietary folic acid during the perinatal period on tissue folate and S-adenosyl-L-methionine (SAM) concentrations and on behavior were examined in 35-day-old DBA/2J mice. In one study dams were started on diets containing no folate (FO), 1.1 mg of folic acid/kg diet (F1.1) or 9.9 mg of folic acid/kg diet (F9.9) on the day of parturition. In a second study some mice were started on the FO or F9.9 diets 6-7 days prior to parturition and some remained on lab chow (LC). The dams and their pups remained on their assigned diets until offspring were killed for biochemical assays. The major findings of the 2 studies are: (1) that eliminating folic acid from diets of dams and developing pups from birth or 1 week prior to birth causes a reduction of folate in brain tissue; (2) that reduction in brain tissue is not as severe as that of liver; (3) that initiating the folate free diet 1 week prior to birth caused reductions in body, liver, and brain weights and in activity levels of surviving offspring; and (4) that offspring of dams started on either the FO or F9.9 diet one week prior to parturition are less viable and have lower levels of SAM in brain tissue than animals reared on the LC diets.


The effect of alpha and beta adrenergic receptor blockers on sleep in the rat. We investigated the effects of the beta adrenergic blocker, propranolol, and the alpha adrenergic blocker, phenoxybenzamine, on sleep patterns in the rat by means of multiple 8-hr and 24-hr polygraphic recordings. Propranolol had no clear effect on time spent in waking, synchronized sleep or desynchronized sleep. Phenoxybenzamine at a dose of 40 mg/kg produced a significant increase in desynchronized sleep time and in the number of desynchronized sleep periods. This is consistent with the view that there may be a mechanism inhibiting the onset of desynchronized sleep periods which involves norepinephrine acting at alpha adrenergic receptors in the brain.


Effects of diazepam and ripazepam on two measures of adjunctive drinking in rats. Four rats were maintained at 85% of their pre-experimental body weights and were given daily 1 hr sessions during which they were each placed in a test chamber in which a 45 mg food pellet was delivered regularly every min independently of behavior. During these sessions water spouts were available to the rats and all 4 animals developed high levels of adjunctive drinking, a burst of licking typically following the consumption of each food pellet. This behavior was found to be sensitive to the effects of diazepam and ripazepam. Small doses of both drugs increased the volume of water consumed during a session. The number of licks was not increased to the same extent, however. Larger doses of both drugs resulted in decreased numbers of licks and decreased water intake although licking appeared on several occasions to be more sensitive than water intake to this action of the drugs. A possible explanation of these effects is that the drugs affected the topography of the rats' licking at the water spouts. Whatever the mechanism involved, however, these results suggest that in such experiments measures of both water intake and number of licks should be obtained.


Scopolamine: effects on fear or defense responses in the rat. In previous research scopolamine reduced fear or defense responses of rats to a cat, and removal of the rats' olfactory bulbs had the same effect. This suggested that scopolamine might have affected defense responses by blocking olfactory perception of the stimulus cat. The present experiments studied this possibility and explored further the effects of scopolamine on defense responses of the hooded rat. In Experiment 1 rats treated with scopolamine were found to be responsive to olfactory cues from a cat. When cat smell, but not a cat, was present in the apparatus, scopolamine-treated rats showed a large and significant suppression of food consumption. In Experiment 2 the effects of scopolamine on defense responses were shown to be generalizable to an inanimate stimulus, mechanical robot. Scopolamine caused significantly less freezing and avoidance and significantly shorter latencies to drink in the presence of the robot. One of the primary findings of the present research is that scopolamine has now been shown to reduce the defensive response of freezing in a variety of stimulus situations. This finding was thought to have important implications for the literature relating anticholinergic drugs and avoidance behavior.


Interactions between naloxone and narcotic analgesics under three schedules that induce polydipsia. A pattern of schedule-induced polydipsia was maintained in rats by a fixed-time schedule where food pellets were presented every 90 sec, a fixed-interval schedule where licking the drinking tube produced pellets every 90 sec, or a fixed-interval schedule where lever presses produced pellets every 90 sec. Under all 3 schedules, injections of morphine, methadone, etonitazene and meperidine generally decreased licking rates and amounts of water consumed, as well as rates of lever-pressing under the schedules where lever presses were required. Naloxone (1 mg/kg) almost completely blocked the effects of morphine and etonitazene, but the effects of methadone sometimes were blocked to a lesser degree. Small increases in the rate of licking and amount of water consumed after the lowest dose of meperidine under the schedule requiring lever-presses were blocked by naloxone, but the higher doses of meperidine that decreased licking, lever-pressing and amount of water consumed under the three schedules were not blocked by naloxone. These data suggest that there are important differences in the ability of naloxone to antagonize the behavioral effects of different narcotic analgesics.


Intracranial injections of 6-OHDA. Comparison of catecholamine-depleting effects of different volumes and concentrations. Fluorescence histochemistry was used to assess monoamine depletion after injections of 6-OHDA into selected brain areas. Two volumes (2 and 4 mul) and 4 concentrations (1, 2, 4 and 8 mug/mul) of 6-OHDA were injected into the olfactory tubercle, the posterior lateral hypothalamus and the lateral hyopthalamus. Selective destruction of catecholamine-containing neurons resulted from all injections of 6-OHDA with the exception of the 2 lowest doses (2 and 4 mul of 1 mug/mul) and the highest dose (4 mul of 8 mug/mul) which produced nonspecific damage of brain parenchyma. The results indicate that, in addition to the selection of an effective dose, it is also possible to choose a site of injection which will produce a maximal area of specific depletion. In cases where injections into terminal areas caused limited specific depletion the same dose injected into preterminal axons often caused a more widespread loss of fluorescence. With volume, concentration and anatomical location being important variables to consider, caution is needed in the interpretation of behavioural experiments. When using 6-OHDA it is necessary to show that specific depletion of catecholamines has been achieved.


A dose-response study of anorectic drug effects on food intake, self-stimulation, and stimulation-escape. A comparison was made of the short-term effects in rats of 3 anorectic drugs (amphetamine, fenfluramine, and phenylpropanolamine) on food intake and responses to obtain brain stimulation and to escape from automatic brain stimulation. At a dose which decreased food intake, amphetamine increased self-stimulation, but not stimulation-escape. Fenfluramine decreased both self-stimulation and stimulation-escape. Phenylpropanolamine, on the other hand, decreased self-stimulation, but not stimulation-escape. Even though all 3 drugs decreased food intake, each of them had different effects on hypothalamic self-stimulation and stimulation-escape. Only the actions of phenylpropanolamine were in agreement with the hypothesis that lateral hypothalamic reward and aversion reflect the animal's tendency to eat, suggesting that other aspects of reinforcement are also involved in lateral hypothalamic stimulation and were affected differently by these drugs.


Acute and chronic single dose effects of LSD-25 on visual discrimination in rats. Rats subjected to either a frontal cortex lesion or to a sham operation were trained to discriminate between a lighted and unlit alley to escape shock. Following intubation with either placebo or LSD-25 (1.0 mg/kg), they were given discrimination trials 2 hr, 1 week, and 1 month later, but with an increased level of task difficulty. Single dose effects of LSD-25 were observed acutely in a transient impairment of visual discrimination accuracy, and more chronically in slower running time. Although no significant drug-lesion interactions were noted, the results were in the direction of a combinatory effect. The value of increasing the level of post-treatment task was confirmed.


Dose-dependent dual effect of morphine on electrophysiologic correlates of positive reinforcement (reward contingent positive variation: RCPV) in the cat. In cats trained to press a lever for milk reward, the postreinforcement EEG synchronization (PRS) and the associated epicortical steady potential shift, known as "Reward Contingent Positive Variation (RCPV) restricted to the occipital cortex, were studied prior to and after administration of morphine sulfate. Contrary to what has ordinarily been described as a typical feline response to morphine, such as restlessness, aggressiveness, rage, and exaggerated startle reaction to environmental stimuli, associated with an increased tonus of the brainstem-hypothalamic arousal system and desynchronized EEG patterns, doses of 0.1-0.4 mg/kg, IM, caused a strong monophasic enhancement of the PRS-RCPV phenomenon. Doses of 0.6-1.0 mg/kg, IM, had clearly a biphasic action: the initial enhancement of the PRS-RCPV responses was followed by their strong suppression. Chlorpromazine (0.1-1.6 mg/kg, i.e.) promptly restored the EEG responses. During the peak effect of the enhancing doses of morphine, the reward-related EEG phenomena also occurred prior to or after the nonrewarded bar press when the animals licked the empty cup. This dissociation of the PRS-RCPV from consumption was much more conspicuous in animals whose control frequency of the PRS oscillations was higher, and after morphine showed more significant slowing. Despite the strong facilitation of the PRS-RCPV in the presence of light, morphine, in contrast to LSD-25, was not able to restore the reward-induced phenomena in the dark.


Effects of catecholaminergic drugs on systems of reward and punishment in experiments on cats. Injection of amantadine or DOPA produced inhibition of both self-stimulation and negative-reinforcing effects of stimulation of the hypothalamus. After injections of l-DOPA in cats pretreated with RO 4-4602, an inhibitor of peripheral decarboxylase, or disulfiram, a dopamine-beta-hydroxylase inhibitor, the inhibitory action on the reinforcing system was enhanced. Amphetamine activated both reward and punishment systems. This data supports an inhibitory function of dopamine in systems of reinforcement and of an activating function of noradrenaline in these systems.