Calcium channel blockade interacts with a neuroleptic to attenuate the conditioning of amphetamine's behavioral effects in the rat. Conditioned responses to drug-related cues appear to be related to the maintenance of stimulant addiction. These conditioned responses are not blocked by treatments that block the direct effects of stimulants and may contribute to the high rate of relapse of addicts. Rats administered (+)-amphetamine in a specific environment exhibit conditioned locomotion when subsequently placed in that environment without drugs. The neuroleptic haloperidol significantly attenuated amphetamine-induced locomotor activity but failed to reduce conditioned locomotion. Nimodipine, an L-type calcium channel antagonist, had no effect on amphetamine-induced unconditioned or conditioned locomotion. However, combined nimodipine and haloperidol treatment blocked the unconditioned and attenuated the conditioned locomotor response to amphetamine. Conjunctive therapy with nimodipine and haloperidol may provide an efficacious treatment for stimulant addiction. In addition, nimodipine may provide an important adjunctive therapy for schizophrenia, allowing the use of lower doses of neuroleptic to avoid extrapyramidal side effects.


Effect of luteinizing hormone-releasing hormone analogs containing cytotoxic radicals on growth of estrogen-independent MXT mouse mammary carcinoma in vivo. Cytotoxic luteinizing hormone-releasing hormone (LH-RH) analogs, AJ-004 (agonist [D-Lys6]LH-RH linked to methotrexate (MTX)), T-98 ([D-Lys6]LH-RH coupled to glutaryl-2-(hydroxymethyl)anthraquinone) (G-HMAQ) and T-121/B (antagonist containing two residues of G-HMAQ) were tested in female BDF1 mice bearing MXT ((3.2)/Ovex) estrogen-independent mammary tumors. All three cytotoxic LH-RH analogs, administered from Alzet Osmotic Minipumps for 3 weeks, produced a significant inhibition of tumor growth. The effects of T-98 and T-121/B were superior to those obtained by treatment with equimolar doses of cytotoxic moiety anthraquinone or the LH-RH carrier alone. We assume that cytotoxic LH-RH analogs have a combined hormonal and cytotoxic activity with a reduced toxicity after administration in vivo. This is the first demonstration of in vivo tumor inhibition by targeted LH-RH analogs bearing cytotoxic radicals.


Subjective and metabolic effects of clodronate in patients with advanced breast cancer and symptomatic bone metastases. Twenty postmenopausal women (aged between 46 and 67 years old) with skeletal metastases from breast carcinoma were treated with clodronate 450 mg i.v. daily for 5 days and thereafter with 100 mg i.m. daily for 10 days. All patients received standard hormonal therapy (tamoxifen). Symptomatic pain (evaluated according to a linear analog scale), performance status (according to Karnofsky), serum alkaline phosphatase, serum creatinine and osteocalcin were measured before and after treatment on days 5, 15, 30 and 45. Scanning by radiology were performed pre- and post-therapy. Bone pain was significantly reduced in 15 out of 20 patients. After clodronate treatment the base line value of circulating osteocalcin (3.2 +/- 1.6 ng/ml) showed a significant increase on days 30 and 45 (p less than 0.001). Radiological assessment of bone lesions showed stable disease in 18 patients and progression in two patients. No adverse side effects were observed. These data show that clodronate provided pain relief in 75% of treated patients and the increase in circulating osteocalcin levels can be considered a marker of the stabilization of skeletal metastatic lesions.


Laparoscopic inguinal herniorrhaphy: classification and 1 year experience. The laparoscopic transabdominal preperitoneal repair of inguinal hernias was utilized in 125 patients in a 12 month period. One hundred and forty three hernias were repaired. These hernias were classified into five different types depending on the site and size of the defect and whether they were primary or recurrent. Cylindrical plugs and screen were used in the first 37 repairs with 2 recurrences and Mushroom Plugs and screen were used in 106 repairs with no recurrences. The two recurrences occurring with the cylindrical plugs were in indirect inguinal hernias with dilated internal rings greater than 2 cm.


Cost effectiveness of laparoscopic cholecystectomy. An investigation was undertaken to determine whether hospital charges for laparoscopic cholecystectomy are higher than those for traditional open cholecystectomy. Thirty consecutive cases of successfully completed laparoscopic procedures in a single surgeon's experience were compared to 30 open cases performed within the previous calendar year. Patients undergoing open cholecystectomy were excluded if coexisting medical problems or complications prolonged hospitalization beyond 7 days. Mean patient age was comparable (open cholecystectomy = 47.3 +/- 2.9, laparoscopic cholecystectomy = 46.5 +/- 2.7 years), as was the incidence of other significant medical problems. Average duration of hospitalization was significantly longer for open cholecystectomy (3.6 = 0.2 days) than for laparoscopic cholecystectomy (1.0 +/- days, p less than .001). Average hospital charges for open cholecystectomy were $5606 +/- 496 and for laparoscopic cholecystectomy $4726 +/- 98. Hospital charges from operating room and recovery room charges alone were $2684 +/- 131 for laparoscopic cholecystectomy and $2196 +/- 113 for open cholecystectomy. These operating room charges represent a significantly higher percentage of total hospital charges for laparoscopic cholecystectomy than open cholecystectomy patients (laparoscopic cholecystectomy = 56.3 +/- 1.9%, open cholecystectomy = 41.2 +/- 1.5%, p less than .05). Average time for return to work or normal activity was significantly shorter for laparoscopic cholecystectomy 8.6 +/- 9 days) than for open cholecystectomy (32.4 +/- 3.6 days, p less than .001). The authors conclude that laparoscopic cholecystectomy is a cost effective procedure for the treatment of symptomatic cholelithiasis, and that increased operative costs more than offset the significantly decreased length of hospitalization.


Cyclosporin A is a hormonal immunomodulator. I. Human study. The immunosuppressant Cyclosporin A (CS) is known to induce 2 hormonal disorders as side effects of its long-term use: gynecomastia in male subjects and hirsutism in female subjects. The present study was started to investigate the problem of whether or not the above steroid-mimetic actions of CS are to be related to the immunosuppressive effect of that substance. Practically, a case-control comparison of 14 urinary steroid excretions was made for each sex with healthy controls and kidney transplant recipients undergoing CS-free and CS-dependent immunosuppressive treatments on the one hand, and the success rate of kidney transplantation was compared between the CS-free treatment group and the CS-dependent treatment group on the other hand. The results obtained are follows: 1) Both the CS-free- and CS-dependent-treatments produced a general depression of all androgens, progestins and corticosteroids in the urine. The suppressive effect on urinary steroid excretions was more marked in the latter than in the former. Under the same CS-free treatment, the excretion of tetrahydrocortisol (THF) was intact in male patients and was significantly depressed in female patients. 2) A remarkable difference between the 2 immunosuppressive treatment groups was found in the ratio of urinary androsterone (AD) to urinary THF, an index of androgen to glucocorticoid balance: the praxis of CS-free treatment significantly depressed log AD/THF in patients of both sexes, the ratio being much higher in normal males than in normal females.(ABSTRACT TRUNCATED AT 250 WORDS)


Initial experience with laparoscopic surgery: establishing a new surgical procedure. Laparoscopic surgery impacted the surgical world in the United States in 1990. This report reviews the initial experience of 34 surgeons in 8 teaching hospitals of the Northeastern Ohio Universities College of Medicine. There were 538 cases reported from May 1, 1990 to January 31, 1991. There was no mortality and the morbidity rate was 4.8%, including three bile duct injuries. The conversion rate to an open procedure was 6.1%. The criteria for credentialing, training, and resident and faculty education are included. The data reported by the Surgery Department of Northeastern Ohio Universities College of Medicine are very similar to reported series from the current literature.


Laparoscopic surgery in the treatment of Mirizzi's syndrome. Mirizzi's syndrome is an unusual complication of cholelithiasis which results when an impacted cystic duct stone causes edema and compression of the adjacent common hepatic duct. The authors report the successful treatment of a patient with this syndrome by laparoscopic means. A flexible 5.0 mm choledochoscope was inserted under laparoscopic guidance into the dilated proximal cystic duct and the impacted stone was extracted. Though still a technically demanding procedure, the authors feel that under certain conditions, laparoscopic surgery may represent the preferred initial approach to patients with Mirizzi's syndrome.


Incidental appendectomy during laparoscopic cholecystectomy. An operative experience of three patients who underwent incidental laparoscopic appendectomy during laparoscopic cholecystectomy is presented. The technique and indications are discussed. The authors conclude that incidental laparoscopic appendectomy is possible and safe with existing incisions performed in gallbladder surgery. However, well-controlled prospective studies should be performed prior to wide application of this technique.


Laparoscopically-assisted hysterectomy for the management of a borderline ovarian tumor: a case report. Borderline ovarian tumors account for 4% of ovarian neoplasms, an incidence which remains constant despite advancing age. Management for younger women can be unilateral oophorectomy, although simple hysterectomy with bilateral salpingo-oophorectomy is more appropriate for women beyond childbearing age. The authors report a laparoscopic approach to a case of borderline ovarian tumor.


Laparoscopic cholecystectomy in situs inversus totalis. A 51-year-old woman with known dextrocardia presented with left-sided abdominal pain and symptoms consistent with biliary colic and cholelithiasis. Abdominal ultrasound confirmed the diagnosis of gallstones, as well as situs inversus with the liver and gallbladder on the left side and the spleen on the right. Laparoscopic cholecystectomy was performed without incident. The procedure was uncomplicated except for being the mirror image of that done with the gallbladder in the normal location. Cholelithiasis occurring with situs inversus is rare and may present a diagnostic problem. The extrahepatic anatomy of the biliary and venous system is the mirror image of the right sided liver. Historic and genetic aspects of situs inversus, as well as current theories regarding its etiology are presented. Situs inversus totalis does not appear to be a contraindication to laparoscopic treatment of cholelithiasis.


An unusual complication of laparoscopic cholecystectomy. Thus far, the morbidity and mortality of laparoscopic cholecystectomy compares favorably to that of open cholecystectomy. Neither procedure, however, is without complications. Laparoscopic cholecystectomy combines diagnostic laparoscopy as well as cholecystectomy. It is the purpose of this article to briefly review these procedures and report an interesting variation: penetration of small bowel as a result of an unusual variation of a Meckel's Diverticulum.


Percutaneous gastrostomy made simple.

Percutaneous gastrostomy made simple. The use of percutaneous endoscopic gastrostomy has obviated the necessity of laparotomy for enteral access. The authors propose a new technique for introduction of the gastrostomy tube. It entails use of the laparoscopic trocar to gain entrance into the gastric lumen.


Laparoscopic preperitoneal prosthetic inguinal herniorrhaphy. Laparoscopic minimally invasive surgical procedures are gaining popularity and becoming more readily available. Presented here is an application of laparoscopy for the repair of groin hernias. Included are actual intraoperative photographs and descriptions of the technique with its rationale for the laparoscopic preperitoneal approach and the use of a prosthetic material.


A simple, safe technique for placement of the veress needle and trocar in laparoscopy. A simple technique of placing two Kocher clamps on the anterior rectus fascia for the elevation of the anterior abdominal wall during the insertion of the veress needle and the laparoscopic trocar is described in 243 consecutive patients. There were no failed insufflations during this study and no cases of preperitoneal emphysema. The technique is simple to use and adds safety to a basically blind procedure.


Placement of drains in laparoscopic procedures. The technique of laparoscopic cholecystectomy is, in many areas, replacing the standard open cholecystectomy as the method of choice. Many surgeons are accustomed to the use of peritoneal drainage following cholecystectomy. The authors have developed a simple, effective, and rapid, yet controlled method of placing drains into the abdominal cavity, using laparoscopic techniques.


The relationship between attitude to disabled siblings and ratings of behavioural competency. The impact of maturation factors and functioning level on the relationship between disabled and non-disabled siblings was examined. Two groups of adults, distinguished by level of functioning of their mentally retarded sibling, completed a Schaefer Sibling Behavior Inventory (SBI) and participated in a semi-structured interview. The latter explored a number of dimensions of the relationship including degree of warmth, contact and involvement. Judgements about involvement and comfort as remembered from the past and judgements about the present were obtained to derive hypotheses about possible changes with time. Data from the SBI indicated that significantly more competent siblings tended to be involved in relationships with a higher degree of reciprocity. In contrast, measures of attitude derived from interview data did not appear to relate to the functioning level of the sibling in any systematic manner. The results suggest that level of discomfort with peers may decrease significantly over the years with neither level of functioning nor gender being important variables. The extent to which non-disabled individuals expressed a preparedness to participate in the life of their disabled sibling may relate to the life-stage of the two. Males in particular anticipated increased future involvement. Although higher-functioning siblings were described as more active in the relationship, this did not appear to influence perceived positiveness of regard. The implications of these results are discussed briefly.


Prevalence of overweight and obesity in Down's syndrome and other mentally handicapped adults living in the community. To find out the prevalence of overweight and obesity in the mentally handicapped, 183 subjects living in the community were studied; 58 of whom had Down's syndrome. It was found that 70.58% of males and 95.83% of females with Down's syndrome, and 49.29% males and 62.96% females from other mentally handicapped subjects, were categorized as overweight and obese, compared with 40% of males and 32% females in that category from normal population.


Lipids and lipoproteins in persons with Down's syndrome. This study was designed to investigate whether the observed decreased prevalence of coronary artery disease in individuals with Down's syndrome may be explained by their serum lipid and lipoprotein profiles. Twenty-seven persons with Down's syndrome and 23 non-affected control individuals were enrolled in this study. Their fasting venous blood was analysed for total cholesterol, triglyceride, LDH cholesterol, HDL cholesterol, apo B and apo AI. The results revealed no significant differences between the study and control group with regard to total cholesterol, LDL cholesterol, apo B and the apo B:apo AI ratio. However, triglyceride levels were significantly increased, and serum HDL cholesterol, apo AI and HDL cholesterol:total cholesterol ratio were significantly decreased in patients with Down's syndrome when compared with the control group. The latter observations are all associated with an increased risk for coronary artery disease. Therefore, it is concluded that the decreased prevalence of coronary artery disease in individuals with Down's syndrome cannot be explained by the lipid and lipoprotein levels observed in this study population.


The role of Wnt genes in vertebrate development. Over the past decade, many potential candidates for molecules involved in pattern formation in the vertebrate embryo have been identified. Manipulation of the expression of some of these factors has generated fascinating results that have allowed investigators to address their roles in embryogenesis. One such family consists of a group of putative cell signaling molecules related to the proto-oncogene Wnt-1. An accumulating body of evidence suggests that the Wnt-family plays a major role in several aspects of vertebrate development.


NMDA receptors in sensory information processing. During the past year electrophysiological studies, particularly in the visual and somatosensory systems, have begun to uncover the specific roles played by NMDA receptors in the processing of sensory information. Many of the features of NMDA-receptor-mediated sensory responses reflect known properties of the receptor.


Chemotherapy-induced acral erythema and acute graft-versus-host disease after allogeneic bone marrow transplantation. Chemotherapy-induced acral erythema is a rare disorder characterized by a painful and intense erythema of the palms and the soles. In allogeneic bone marrow transplant patients, the differential diagnosis of acute graft-versus-host disease (AGVHD) may be difficult. We describe a case of concurrent acral erythema and AGVHD. The clinical features of both conditions as well as the histological findings on serial skin biopsy specimens are discussed.


Impaired suppressor cell activity due to surface sulphydryl oxidation in rheumatoid arthritis. Rheumatoid arthritis is a chronic inflammatory disorder with considerable evidence of impaired regulation of the immune response, including defective suppressor cell function, especially in the synovial membrane. We have investigated whether oxidation of cell surface thiols might be responsible for these defects and whether such cell function may be modulated towards normal by treatment with a sulphydryl-reactive drug, D-penicillamine. Using healthy mononuclear cells treated with an impermeant thiol blocker, induction of suppressor activity by incubation with the lectin Con A was not dependent on surface sulphydryl groups but suppressor activity was abolished by thiol blockade after Con A stimulation. Peripheral blood mononuclear cells from patients with active rheumatoid disease showed impaired Con A-induced suppressor activity which was enhanced to near-normal levels by incubating the rheumatoid cells with a sulphydryl reducing agent, 2-mercaptoethanol, or D-penicillamine. Con A-stimulation of cells from patients treated with intramuscular gold or D-penicillamine generated more active suppression than those from patients receiving non-steroidal drugs only. Mononuclear cells from patients with other chronic inflammatory joint diseases showed normal Con A-induced suppressor activity. These data support the conclusion that surface thiols on mononuclear cells in rheumatoid arthritis are reversibly oxidized by the disease process. This gives rise to aberrant cell function including impaired suppressor activity. Such a mechanism may be at least partly responsible for the defective immunoregulation seen in rheumatoid patients and thus be a relevant target for thiol containing antirheumatic drugs.


Evidence that the lateral tegmental field plays an important role in the central sympathoinhibitory action of 8-OH-DPAT. We examined the effects of 8-OH-DPAT and 5-HT on three types of sympathetic-related neurons identified in the lateral tegmental field of anesthetized cats using spike-triggered averaging techniques. Based on their response to baroreceptor activation, these neurons could be classified as sympathoexcitatory, sympathoinhibitory, or baroreceptor activation unresponsive. 8-OH-DPAT administered intravenously was found to inhibit sympathoexcitatory neurons with a high degree of correlation to inhibition of sympathetic activity, and to excite sympathoinhibitory neurons in a dose-dependent manner. Iontophoretic application of 8-OH-DPAT and 5-HT to the majority of sympathoexcitatory neurons caused inhibition of spontaneous activity while iontophoretic application of 8-OH-DPAT to sympathoinhibitory neurons had variable effects although 5-HT consistently caused excitation. Baroreceptor unresponsive neurons were insensitive to iontophoretic 8-OH-DPAT and showed only limited response to 5-HT. It is concluded that the lateral tegmental field plays an important role in the sympathoinhibitory action of 8-OH-DPAT.


Amygdala kindling does not alter the N-methyl-D-aspartate receptor-channel complex which modulates dopamine release in the rat striatum and amygdala. Kindling is suggested to be critically associated with enhancement of N-methyl-D-aspartate (NMDA) type excitatory amino acid synaptic transmission. The present study examined effects of kindling on NMDA-induced dopamine (DA) efflux from slices of the rat striatum and amygdala. When assayed 5-7 days after the last evoked seizure, no difference was observed between kindled and non-kindled striatum in the ability of NMDA to induce DA release, or in the effect of MK-801 or 7-chlorokynurenic acid to inhibit the amino acid-induced transmitter release. The present study revealed that NMDA receptors are also involved in the modulation of DA release in the amygdala. However, no difference was observed between kindled and non-kindled amygdala in the ability of NMDA to induce DA release. These results suggest that amygdala kindling does not alter activity of the NMDA receptor-channel complex modulating DA release in the striatum and amygdala.


Effects of RPE-cell factors secreted from permselective fibers on retinal cells in vitro. This study was undertaken to determine if retinal pigment epithelial (RPE) cells encased in permselective hollow fibers survive in a tissue culture environment and secrete a diffusible trophic factor(s) that may affect retinal cell survival in vitro. In this study, RPE cells were isolated from 6- to 8-day-old Long-Evans rats, then loaded into hollow fibers. The RPE-cell fibers were then cultured for at least one week in serum-containing medium. These RPE-cell fibers were subsequently co-cultured with cells isolated from retinas of day 2 Long-Evans rats in a defined medium. For at least 6 days in culture, opsin-positive cells were observed on the surface of larger flat cells. Over 80% of the small, round cells immunostained for opsin. However, opsin-immunostained cells were seldom seen in cultures with control fibers, that lacked RPE cells. In addition, conditioned medium collected from either the RPE-cell fibers or cultured RPE cells affected survival of opsin-positive retinal cells in culture in a manner similar to that of the RPE-cell fibers. Furthermore, selected growth factors such as epidermal, nerve and fibroblast growth factors, were unable to sustain retinal cell survival and affect morphological development as seen in RPE-CM supplemented cultures. In vivo companion developmental studies demonstrated that few opsin-positive cell bodies were observed in retinas of day 2 Long-Evans rats, the age corresponding to the stage of retinal cell isolation. In retinas of day 5 Long-Evans rats, the age corresponding to the end point of the in vitro assay, a dramatic increase in the number of opsin-immunostained cell bodies was noted, which corresponds to the developmental sequence also seen in culture. Light and electron microscopic examination revealed that the RPE cells cultured in the hollow tubes maintained an RPE-like structure for several months, in that these cells contained melanosomes and extended microvilli from their apical border and formed junctional complexes with adjacent cells. Results of this study confirm our earlier findings that RPE cells secrete an apparent novel factor(s) that affects retinal cell survival in vitro and, most significantly, the described encapsulation/secretion mechanism may provide a convenient method to deliver such factors for further in vivo testing of this phenomenon.


Interleukin-1 receptor antagonist inhibits ischaemic and excitotoxic neuronal damage in the rat. Interleukin-1 (IL-1) synthesis in the brain is stimulated by mechanical injury and IL-1 mimics some effects of injury, such as gliosis and neovascularization. We report that neuronal death resulting from focal cerebral ischaemia (middle cerebral artery occlusion, 24 h) is significantly inhibited (by 50%) in rats injected with a recombinant IL-1 receptor antagonist (IL-1ra, 10 micrograms, icv 30 min before and 10 min after ischaemia). Excitotoxic damage due to striatal infusion of an NMDA-receptor agonist (cis-2,4-methanoglutamate) was also markedly inhibited (71%) by injection of the IL-1ra. These data indicate that endogenous IL-1 is a mediator of ischaemic and excitotoxic brain damage, and that inhibitors of IL-1 action may be of therapeutic value in the treatment of acute or chronic neuronal death.


[Early intensification of treatment in diabetes using a combination of insulin and nicotinamide--effect on C-peptide levels and the course of the disease]. Significantly higher C-peptide levels were found in a group of 49 juvenile diabetics treated with insulin in combination with nicotinamide compared to 33 diabetics treated with insulin only. There had been no significant differences in the initial values of C-peptide between the two groups. After six weeks the mean basal values of C-peptide in the blood of nicotinamide treated diabetics reached 1.21 +/- 0.79 ng/ml compared to 0.87 +/- 0.55 ng/ml determined in the group treated with insulin only (p less than 0.05). After one year of therapy the differences remained significant (p less than 0.01). The insulin requirement was significantly lower in the nicotinamide treated group after six weeks (p less than 0.001) and the difference remained significant also after two years of nicotinamide treatment (p less than 0.02). A longer period of partial remission was recorded in the group of diabetics who were administered nicotinamide. (Tab. 4, Fig. 2, Ref. 13.)


Efficacy of ondansetron against nausea and vomiting caused by dacarbazine-containing chemotherapy. The antiemetic activity of ondansetron (Zofran, Glaxo Pharmaceuticals, Research Triangle Park, NC) was evaluated in 25 patients with recurrent melanoma who were treated sequentially with dacarbazine (DTIC), vinblastine, and cisplatin. The antiemetic regimen included ondansetron alone in 11 patients; ondansetron plus lorazepam (Ativan, Wyeth-Ayerst, Philadelphia, PA) in 9 patients; and ondansetron plus lorazepam plus metoclopramide (Reglan, A. H. Robins Co., Richmond, VA) in 5 patients. Twenty-one patients had no prior exposure to chemotherapy, whereas 4 patients had previously received the same chemotherapy regimen and had severe vomiting despite administration of standard antiemetics. The antiemetic efficacy of ondansetron was impressive. Administration of a single dose of 10 mg resulted in complete control of nausea and vomiting in 22 patients, and the remaining 3 patients had only mild vomiting. Ondansetron is highly effective in controlling the nausea and vomiting caused by dacarbazine.


Double-blind, parallel-group evaluation of etodolac and naproxen in patients with acute sports injuries. The efficacy and safety of etodolac and naproxen were compared in a double-blind, randomized, parallel-group outpatient study. Patients with acute sports injuries were assigned to receive either etodolac 300 mg TID (50 patients) or naproxen 500 mg BID (49 patients) for up to 7 days. Assessments were made at the pretreatment screening (baseline) and at days 2, 3, 4, and 7 of treatment. Assessments included patient and physician global evaluations, spontaneous and induced pain intensity, range of motion, tenderness, heat, degree of swelling, and degree of erythema. Safety assessments, including laboratory profiles, were made at pretreatment and at final evaluation; patients' complaints were elicited at all visits. Both treatment groups showed significant (P less than or equal to 0.05) improvement from baseline for all efficacy parameters by day 2 and thereafter at all time points. Improvement was similar for the two groups. No patients in either group withdrew from the study because of drug-related adverse reactions. The results of this study indicate that etodolac (900 mg/day) is effective and well tolerated as an analgesic and anti-inflammatory in acute sports injuries and is comparable to naproxen (1000 mg/day).


Safety and efficacy of etodolac compared with piroxicam in patients with degenerative joint disease of the knee. The efficacy and safety of etodolac and piroxicam were compared in a double-blind, randomized, parallel-group outpatient study at four sites. Patients with active osteoarthritis of the knee were assigned to receive etodolac 600 mg/day (57 patients) or piroxicam 20 mg/day (59 patients) for 6 weeks. Efficacy assessments were made at the pretreatment screening, at baseline, and at treatment weeks 2, 4, and 6 for patient and physician global evaluations, night pain, spontaneous pain intensity, weight-bearing pain variables, measures of inflammation, morning stiffness, and knee flexion. An analysis was also done based on each patient's final evaluation, regardless of the week at which it occurred. Safety assessments were made before treatment and at the completion of therapy. A therapeutic response was obtained in both treatment groups by the end of the second week of treatment. At the final evaluation, both groups showed significant improvement (P less than or equal to 0.05) from baseline for most efficacy assessments. The physician's global assessment indicated improvement in the condition of 60% of the etodolac-treated patients and 39% of the piroxicam-treated patients at the final evaluation. There was no significant difference between treatment groups in the number of patient withdrawals due to adverse reactions or in the number of patients reporting side effects. The results of this study indicate that, compared with piroxicam 20 mg/day, etodolac 600 mg/day is effective and well tolerated in the treatment of patients with osteoarthritis.


Bioavailability of medroxyprogesterone acetate from three oral dosage formulations. The bioavailability of three formulations of medroxyprogesterone acetate (MPA) was assessed in 30 healthy male volunteers in a three-way, open-label, cross-over-designed trial. Each subject received one Provera 500-mg tablet, one Farlutal 500-mg tablet, and one Provera 500-mg granule packet according to a randomized schedule, with each treatment separated by a 21-day washout period. Serum MPA levels were determined using both radioimmunoassay (RIA) and high-performance liquid chromatography techniques. Based on the results of RIA analysis, Farlutal tablets produced significantly lower serum MPA concentrations compared with Provera tablets at most sampling times, resulting in statistically lower AUC0-144 for the Farlutal tablet (544 vs 768 ng.hr/ml; -29.2%). The Farlutal tablet also had a significantly lower maximum concentration than the Provera tablet (27.8 vs 47.4 ng/ml; -41.4%). However, there was no significant difference in time of maximum concentration between the tablet formulations (3.71 vs 3.41 hr), indicating that the rates of absorption of the two tablet formulations were comparable. Provera granules provided significantly higher serum MPA levels than Provera tablets at 0.5, 1, 1.5, 2, and 6 hours, and the AUC0-144 for Provera granules was higher by 5.47% (810 vs 768 ng.hr/ml). There were no differences in terminal elimination rate constants among the dosage forms. No significant adverse events were noted during the trial. The relative bioavailabilities of Provera granules and Farlutal tablets were 105% and 71.2%, respectively, compared with Provera tablets.


Immunological alterations in cluster headache during remission and cluster period. Comparison with low back pain patients. Cluster headache is a disorder of unknown origin. Some studies have focused their attention on neuroendocrine derangement, others on immunity. To probe central alterations in cluster headache (CH), immune parameters were investigated in cluster headache patients in comparison to low back pain patients and healthy controls. Increases in peripheral blood monocytes found in remission cluster headache patients may be attributable to chronic central nervous system (hypothalamic?) noradrenergic dysfunction or altered beta-endorphin. Alterations in NK+, CD3+ and CD4+ levels found in cluster period cluster headache and low back pain patients are probably pain or stress-related.


Regulation of retinoblastoma protein functions by ectopic expression of human cyclins. The retinoblastoma susceptibility gene (RB) product, the retinoblastoma protein (pRb), functions as a regulator of cell proliferation. Introduction of the RB gene into SAOS-2 osteosarcoma cells, which lack functional pRb, prevents cell cycle progression. Such growth-suppressive functions can be modulated by phosphorylation of pRb, which occurs via cell cycle-regulated kinases. We show that constitutively expressed cyclins A and E can overcome pRb-mediated suppression of proliferation. pRb becomes hyperphosphorylated in cells overexpressing these cyclins, and this phosphorylation is essential for cyclin A- and cyclin E-mediated rescue of pRb-blocked cells. This suggests that G1 and S phase cyclins can act as regulators of pRb function in the cell cycle by promoting pRb phosphorylation.


Apoptosis, cell proliferation and c-ras expression during and after cyproterone acetate (CPA) induced liver hyperplasia. Cell proliferation and cell death appear in several systems as mutually exclusive, which raises the assumption that a same factor or secondary signal(s) might exert opposite control on the two processes. To test this assumption we investigated the time-course evolution of the S phase and apoptotic indices in rat liver during cyproterone acetate (CPA) induced hyperplasia and during the recovery of normal liver mass provoked, respectively, by cyproterone acetate (CPA) treatment and withdrawal. The levels of c-myc and c-ras transcripts were also followed in view of the indications of a positive role of these oncogenes in proliferation. The data showed that proliferation and cell death are not always mutually exclusive and that a high rate of cell death was indifferently associated with high or low c-ras expression. Our data are consistent with a role of this gene in proliferation but exclude that it plays an opposite role in controlling cell death.


[Genotypes of Neisseria meningitidis strains isolated from patients in the Czech Republic]. A group of 75 strains of Neisseria meningitidis isolated from patients with meningococcal meningitis in the Czech Republic during 1980-1988 was characterized by assessment of serogroups, serotypes, serosubtypes and genotypes. Twelve strains belonged into serogroup A, 27 into serogroup B and 36 into serogroup C. The most frequently found serotype was 4, subserotype P1.2. The authors identified 48 different enzyme genotypes among which four genetically related groups were assessed. Strains of Neisseria meningitidis of the same genotype were found more frequently during the period of 1980-1984, when a rise of meningococcal meningitis in the Czech Republic was recorded. During this period 73% of the strains belonged into three genetically related groups, while during the subsequent period (1985-1988) these genetically related groups comprised only 47% of the strains. Only one strain of Neisseria meningitidis (878/85) represented a genetic clone ET-5 complex responsible for an epidemic of meningococcal meningitis in western European countries from the mid-seventies.


[Levels of C-reactive protein in serum--comparison of 2 methods used for quantitative determination]. The authors compared the results of two methods used for the quantitative assessment of C-reactive protein: the method of radial immunodiffusion and the microturbidimetric method. The testing was based on examination of 68 sera from patients. The samples comprised sera with a CRP concentration under 230 mg/l. There was a statistically significant correlation between the results of the two compared methods (r = 0.949, p = 0.0001).


[Identification of enteric bacteria using the ENTEROtest 1 and 2 and the ENTERO-Rapid systems]. The identification efficacy of two systems, ENTEROtest 1 & 2 and ENTERO-Rapid (fy. Lachema a. c., Brno), was compared. A total 123 well known strains of enteric bacteria were tested. The ENTEROtest 1 & 2 system correctly identified 87.0% tested strains to the species level, the ENTERO-Rapid system correctly identified 76.4% of these strains.


[Diagnosis of visceral mycosis].

[Diagnosis of visceral mycosis]. The author describes techniques of laboratory processing of different biological materials used for detection of potentially pathogenic micromycetes which are involved in the aetiology of secondary mycoses. The author describes in detail methods for the quantitative assessment of yeasts in urine and sputum.


[Plasmid profile and toxicity of Klebsiella pneumoniae strains isolated at a neonatal department]. The authors tested the plasmid profile of 11 and the toxicity of 16 strains of K. pneumoniae isolated in a neonatal department. Based on examination of the plasmid profile it may be assumed that very probably a nosocomial spread of the strains was involved. This is suggested by the identical plasmid profile of all strains isolated in the environment and five strains isolated from neonates. On investigation of the enterotoxicity of culture filtrates of the investigated strains the authors did not reveal the presence of a thermolabile or thermostable enterotoxin. In the skin test on rabbits 93.8% of the tested samples were positive. Morphological changes on Vero cells after 48 hours action were produced by four culture filtrates. Only two strains caused haemolysis of agar with 6% sheep and rabbit blood after incubation at 37 degrees C. Another 14 strains caused lightening of the agar with rabbit blood after subsequent incubation at 4 degrees C.


[Diabetes and viral hepatitis B].

[Diabetes and viral hepatitis B]. The author emphasizes the fact that diabetics are also in the Czech Republic a group with a high risk of infection with viral hepatitis type B (VHB), i. e. in diabetics a higher prevalence of acute VHB is found, a higher prevalence of HBsAg and anti-HBs, and in diabetics chronic sequelae acute hepatitis are more frequent. The author recommends active immunization against VHB and to focus attention on the group most at risk, i. e. to examine in the Czech Republic all diabetics treated with insulin for the presence of HBsAg and anti-HBs and to immunize subsequently against VHB negative subjects from this group.


[The health impact of stray and wild cats in the human environment]. The population of stray cats in towns expands due to irresponsible behaviour of humans who allow them to enter cellars, hot water canals, store rooms etc. As a result of frequent feeding some specimens become tame, lose their shyness and on contact with humans and domestic animals they can become the source of some infectious agents. The most important of thus transmitted diseases is rabies. Cats become infected from foxes on rubbish heaps on the periphery of communities. More than 60% of cases of rabies of domestic animals are found in cats. Toxoplasmosis--cats excrete the infectious stage of the causal agent in their excreta. In the same manner also the agent causing intestinal salmonelloses is excreted. More than 5% cats are infected. Via excreta also the causal agents of toxocariasis are spread--the larvae of cat thread worms settle in the internal organs of humans (larva migrans). The assumed incidence in cats is as high as 50%, as suggested also by the high positivity of specimens from children playgrounds. Stray cats may be the source of agents causing various dermatomycoses and of ectoparasites.


[Immunologic cross reactivity of beta-lactam antibiotics]. The authors investigated the antibody response against beta-lactam antibiotics (penicillins, cephalosporins, monolactams, carbapenes) in conjugation with allergic reactions after penicillin administration. The results revealed a high percentage of positive antibody responses against penicillin and high percentage of crossed immunological reactivity between penicillin and the other investigated preparations.


[Significance of hybridization methods in microbiological diagnosis: DNA probes and PCR]. The authors give an account of hybridization methods, in particular DNA probes and the polymerase chain reaction (PCR) and their application in microbiological diagnosis. They deal with the principle of the two methods and their applications in the diagnosis not only of genera and species but also different factors of pathogenicity and virulence. The authors mention the advantages and disadvantages of radioactively and non-radioactively labelled probes such as PCR. They draw attention to the perspective application of these methods in research and diagnostic laboratories.


A single amino-acid substitution in the beta-tubulin gene of Neurospora confers both carbendazim resistance and diethofencarb sensitivity. Two MBC-resistant mutants of Neurospora crassa, F914 and F939, were sensitive to diethofencarb at a concentration of 0.1 micrograms/ml, while the wild-type strain and other MBC-resistant mutants showed resistance to diethofencarb at a concentration of 100 micrograms/ml. Genetic analysis suggested that the mutations in these two strain were closely linked to the Bml locus which codes for beta-tubulin. When the wild-type strain was transformed by the cloned beta-tubulin gene of the F914 strain, the transformants showed both MBC resistance and diethofencarb sensitivity. On the other hand, the diethofencarb sensitivity of the F914 strain was cancelled by transformation with the wild-type beta-tubulin gene. DNA sequencing of F914 beta-tubulin revealed that glycine was substituted for glutamic acid at position 198 in the F914 strain. Therefore, a single base change in the beta-tubulin gene was proved to confer both MBC resistance and diethofencarb sensitivity.


Expression of Neurospora crassa laccase under the control of the copper-inducible metallothionein-promoter. Laccase from the ascomycete Neurospora crassa is an inducible secretory enzyme. In vegetatively growing cultures its biosynthesis is repressed but can be induced by different protein synthesis inhibitors. Transformation of the N. crassa wild-type strain Singapore with a fusion gene consisting of the N. crassa copper-metallothionein promoter and the laccase gene are described in this report. Correct integration of the 3.6 kilobase (kb) promoter-fragment fused with the laccase gene containing a 5' consensus region leads to copper-dependent expression of the enzyme during the vegetative growth phase. The enzyme is glycosylated and secreted, and high amounts of extracellular activity can be detected. The regulation of laccase biosynthesis of one examined transformant, followed at both the transcriptional and the translational level, indicates co-induction of both copper-metallothionein and laccase. The data presented show that expression of the recombinant laccase gene is exclusively regulated by the transformed N. crassa metallothionein-promoter.


Transformants of Neurospora crassa with the nit-4 nitrogen regulatory gene: copy number, growth rate and enzyme activity. nit-4 is a pathway-specific regulatory gene which controls nitrate assimilation in Neurospora crassa, and appears to mediate nitrate induction of nitrate and nitrite reductase. The NIT4 protein consists of 1090 amino-acid residues and possesses a single GAL4-like putative DNA-binding domain plus acidic, glutamine-rich, and polyglutamine regions. Several mutants with amino-acid substitutions in the putative DNA-binding domain and a nit-4 deletion mutant, which encodes a truncated NIT4 protein lacking the polyglutamine region, are functional, i.e., they are capable of transforming a nit-4 mutant strain. However, transformants obtained with most of these nit-4 mutant genes possess a markedly reduced level of nitrate reductase and grow only slowly on nitrate, emphasizing the need to examine quantitatively the affects of in vitro-manipulated genes. The possibility that some mutant genes could yield transformants only if multiple copies were integrated was examined. The presence of multiple copies of wild-type or mutant nit-4 genes did not generally lead to increased enzyme activity or growth rate, but instead frequently appeared to be detrimental to nit-4 function. A hybrid nit-4-nirA gene transforms nit-4 mutants but only allows slow growth on nitrate and has a very low level of nitrate reductase.


[Huntington's chorea from the viewpoint of forensic psychiatry]. In the initial stages Huntington's chorea is rarely correctly diagnose. The patient however in the early period of the illness is often subjected to social rejection and is more likely to be the victim of criminal acts or the perpetrator of such acts. The article with 3 cases studies concentrates attention to the need to take Huntington's chorea into consideration in determining psychiatric and psychological aspects of tribunals.


Effect of freezing and thawing of serum on the immunoassay of lipoprotein(a). We studied the effect of freezing and thawing of serum on the determination of lipoprotein(a) [Lp(a)] with a commercial enzyme-linked immunosorbent assay (ELISA) and an immunoturbidimetric assay (ITA). Portions of sera from 11 apparently healthy persons and pooled sera, from an additional 10 subjects were frozen at either -20 or -70 degrees C and thawed at room temperature. Cycles of freezing and thawing were repeated during the experiments (1 month). Samples were assayed for Lp(a) after thawing. Pooled sera were subjected to quick freezing at -70 degrees C and thawing at room temperature in cycles. Results show a significant (P less than 0.05) decrease in Lp(a) concentration in sera subjected to freezing and thawing. Samples thawed from -20 degrees C gave concentrations by ELISA that were significantly lower than those of fresh samples after one freeze-thaw cycle. By ITA the decrease was significant only after two cycles. In specimens frozen at -70 degrees C, Lp(a) concentrations determined by ELISA decreased after two cycles, and by ITA after three freeze-thaw cycles. Serum samples subjected to quick freezing at -70 degrees C and thawing did not show significant decreases in Lp(a) immunoreactivity during four cycles. Immunoreactivity of Lp(a) in samples stored at 4 degrees C decreased after 6 days but fell faster in serum samples subjected to freezing and thawing before storage at 4 degrees C.


Comparison of assay kits for unconjugated estriol shows that expressing results as multiples of the median causes unacceptable variation in calculated risk factors for Down syndrome. We compared the performance of two methods for assaying unconjugated estriol in serum: the modified Amerlex third-trimester RIA kit, as used in seminal papers on unconjugated estriol in Down syndrome screening, and the new optimized Amerlex-M second-trimester kit. The significant difference between the results of each assay could cause unacceptable changes in the detection rate and false-positive rate of Down syndrome screening programs, especially if previously published values for estriol are used in the risk calculation. It is not possible to define new calculation parameters for every assay kit because new parameters will need to be defined every time kit changes occur, which would require a large collection of samples from Down syndrome pregnancies for standardization. Possible solutions to this problem are discussed.


Rat plasma clearance rate and organ distribution of beta-hexosaminidase isoenzymes from human serum. beta-Hexosaminidase (EC 3.2.1.30) is markedly increased in human serum in liver disease, chronic alcoholism, and pregnancy. Knowledge of the clearance rate of plasma/serum beta-hexosaminidase is necessary to evaluate this increase. We studied the plasma clearance of beta-hexosaminidase isoenzymes (purified from human serum and placenta) after their infusion into rat circulation. A recently developed enzyme immunoassay method was used to measure the human beta-hexosaminidase isoenzymes; this method relies on both immunoreactivity and enzyme activity, so intact human isoenzymes were measured. In comparison with the placental isoenzymes (t1/2 less than 2 min), the serum forms showed a considerably slower clearance (t1/2 = 2-4 h). However, desialylation of the serum forms resulted in their rapid clearance (t1/2 less than 2 min). The organ localization of the enzyme eliminated from the circulation was investigated 24 h after infusion. Placental and native serum isoenzymes accumulated mainly in the liver and the spleen. Desialylated serum forms were almost exclusively localized to the liver.


Amlodipine versus nifedipine retard in the treatment of chronic ischemic heart disease. The efficacy of amlodipine, a long half-life dihydropyridine calcium antagonist, at the dosage of 5-10 mg/day in a single daily administration, has been compared with that of nifedipine R, a short half-life dihydropyridine, at the dosage of 20-40 mg b.i.d. in 29 patients with chronic ischemic heart disease. After a one week placebo period, patients were assigned to the treatment with amlodipine or nifedipine R, according to a randomized sequence and a cross-over, single-blind design, for two control periods of four weeks and without a wash-out interval between these two phases. During the stress test, a significant increase from baseline in test duration and in time to onset of ischemia and of angina have been obtained with both treatments; moreover amlodipine increased significantly the time to onset of ST segment deviation (-1 mm) and the time to maximum ST segment deviation compared with nifedipine R changes. Also with Holter monitoring and in the angina diary there was a significant reduction of anginal episodes. As regards safety profile, amlodipine treatment was associated with a significantly lower incidence of side effects compared with nifedipine R. This is probably due to the particular pharmacokinetics of amlodipine which, besides the long half-life which allows a single daily administration, shows a retarded peak (between the 6th and the 12th hour) with consequent reduction of phenomena connected with fast and excessive peripheral vasodilatation. In conclusion, amlodipine was as effective in reducing the signs of ischemia as nifedipine R, but compliance was better due to the single daily administration and so was tolerability.


Different sensitivity of the sarcoplasmic reticulum Ca(2+)-ATPase enzyme to fluorescein-isothiocyanate in rabbit and carp muscles. 1. Carp and rabbit sarcoplasmatic reticulum Ca(2+)-ATPase enzymes were compared with respect to their sensitivity to FITC labelling. 2. The carp enzyme showed much lower sensitivity to FITC in the Ca(2+)-Mg2+ activated ATPase activity. Fifty percent inhibition was observed at 20 microM labelling FITC concentration; in rabbit enzyme this inhibition was already achieved at 2 microM FITC. 3. The tryptic cleavage products of the carp enzyme identified with immunoblot analysis as well as with FITC fluorescence, suggest multiple cleavage, yielding different fragments from the ones well known in rabbit and in rat enzyme. 4. The present results indicates major structural differences with respect to the FITC binding, and tryptic cleavage between the SR Ca(2+)-ATPase enzymes from carp and rabbit, despite the cross-reactivity with polyclonal antibodies.


Evaluating retinal circulation using video fluorescein angiography in control and diabetic rats. Video fluorescein angiography has been used to evaluate retinal circulatory parameters in diabetic and non-diabetic Sprague-Dawley rats. Video fluorescein angiograms were recorded from the retina using a modified retinal fundus camera following a 5 ul bolus injection of sodium fluorescein dye into the jugular vein. Retinal circulatory parameters were measured using computer assisted image analysis. These analyses were performed on 25 diabetic rats with 1 week duration of diabetes and 26 matched, non-diabetic, rats. There was a significant (p = .0001) increase in retinal Mean Circulation Time (MCT) in the diabetic group (1.83 +/- 0.40 s) compared to the control group (1.09 +/- 0.27 s). There were no significant differences in arterial or venous diameters comparing diabetic and control groups. In a separate paired experiment, measurements were made from the same animals both before and after one week duration of diabetes. A paired t-test analysis demonstrated significantly increased MCT times in the 6 diabetic animals (p = .001) while there was no significant differences detected in the 4 corresponding control animals. These results indicate that significant increases in retinal circulation times can be measured as early as 1 week after streptozotocin induced diabetes in this animal model.


Immunohistochemical localization of basement membrane components in pseudoexfoliation material of the lens capsule. The pseudoexfoliation (PSX) syndrome has long been speculated to be a disorder of disturbed basement membrane metabolism. To test this concept, we investigated the presence of all principal basement membrane components in precapsular PSX deposits of 30 anterior lens capsules by immunofluorescence and electron microscopic immunogold techniques. We have shown heparan sulfate and chondroitin sulfate proteoglycans, laminin, entactin/nidogen, fibronectin, and amyloid P protein to be an integral constituent of PSX material; type IV collagen was restricted to a microfibrillar layer interposed between capsular surface and typical PSX material. The additional presence of elastin epitopes indicates that the PSX material is a multicomponent expression of a disordered extracellular matrix synthesis including the incorporation of the principal noncollagenous basement membrane components. The extensive labelling of PSX material for chondroitin sulfate suggests an overproduction and abnormal metabolism of glycosaminoglycans to be one of the key changes in this disorder.


Biological profile of the metabolites and potential metabolites of the anticonvulsant remacemide. Remacemide hydrochloride ((+/-)-2-amino-N-(1-methyl-1,2-diphenylethyl)- acetamide hydrochloride or FPL 1292AA) is a novel compound undergoing clinical trials for patients with generalized tonic/clonic and complex partial epilepsy. Remacemide exhibits efficacy against maximal electroconvulsive shock (MES) in rodents and seizures elicited by N-methyl-D,L-aspartate (NMDLA) in mice. Using rat synaptic membrane fractions, remacemide was shown to possess relatively weak noncompetitive binding to the ionic channel site of the NMDA (N-methyl-D-aspartic acid) receptor complex. With the hypothesis that activity against NMDLA-elicited seizures might be reflected by transformation to a more active metabolic species, the aim of the present study was to evaluate potential pharmacological effects of the 9 identified metabolites of remacemide which were all found in human and dog urine. Moreover, specific entities were recognized in plasma (including the rat's), as well as dog and rat cerebrospinal fluid. Five putative metabolites were also examined. A major route of metabolic transformation of remacemide in rats yields the formation of a pharmacologically active more potent desglycine derivative, namely FPL 12495 (+/-). Potency over the parent compound is revealed in the MES test in mice and rats, the NMDA-induced convulsions/mortality test in mice, and especially involving in vitro displacement of MK801 binding to the channel subsite of the NMDA receptor. The S isomer (FPL 12859) of this desglycinate is even more potent, while the R isomer is less potent than the corresponding racemate. Unlike the non-competitive NMDA antagonist, MK801, these desglycinates did not prevent kindled seizures. Three other identified metabolites show efficacy in the mouse and rat in vivo tests, namely the N-hydroxy-desglycinate (FPL 15053) and the p-hydroxy-desglycinates (FPL 14331 and FPL 14465). FPL 15053 exhibited modest activity in all tests. The only in vivo activity exhibited by the 2 p-hydroxy-desglycinates was evidenced in the MES test following i.p. and i.v. dosing. However, FPL 14331 was active in the MK801 binding assay. An oxoacetate metabolite, PFL 15455, failed to demonstrate any biological activity. Of potential metabolites tested 2 beta-hydroxy-desglycinates (FPL 14991 and FPL 14981) displayed modest activity in the MES test, however, only FPL 14981 prevented NMDLA-induced convulsions/mortality in mice and was 2-fold more active regarding MK801 binding. The hydroxy-methyl derivative of remacemide (FPL 13592) and its desglycinate (FPL 15112) prevented MES-induced convulsions only after i.v. administration; only the desglycine derivative displaced MK801 binding.(ABSTRACT TRUNCATED AT 400 WORDS)


Teicoplanin plus ciprofloxacin versus gentamicin plus piperacillin in the treatment of febrile neutropenic patients. Teicoplanin plus ciprofloxacin was compared with gentamicin plus piperacillin for the empirical treatment of fever in 80 neutropenic patients. A favourable response was seen in 78% of patients receiving teicoplanin plus ciprofloxacin and in 49% receiving gentamicin plus piperacillin (p less than 0.05). When microbiologically documented episodes were analysed separately, the response to teicoplanin plus ciprofloxacin was favourable in 81% of patients whereas only 35% responded favourably to gentamicin plus piperacillin (p = 0.034). Gram-positive organisms accounted for 76% of bacterial isolates, Staphylococcus epidermidis being the most common pathogen. Ten of 12 (83%) Staphylococcus epidermidis infections resolved when treated with teicoplanin plus ciprofloxacin as compared with 2 of 8 (25%) treated with gentamicin plus piperacillin. Teicoplanin is at least as effective as gentamicin plus piperacillin in the empirical treatment of febrile neutropenic patients and may be more effective in situations where gram-positive organisms are prevalent. The high incidence of gram-positive infections in our unit justifies the use of an agent with specific activity against gram-positive organisms in the first-line antibiotic regimen.


Effect of amperozide on rat cortical 5-HT2 and striatal and limbic dopamine D2 receptor occupancy: implications for antipsychotic action. Amperozide (FG 5606, N-ethyl-4-[4',4'-bis(p-fluorophenyl)butyl]-1-piperazine-carboxamide) is an atypical antipsychotic drug which has relatively weak in vitro affinity for striatal dopamine2 (D2) receptors and strong affinity for cortical 5-HT2 receptors. The in vivo affinity for 5-HT2 binding sites in rat cortex was 1.1 mg/kg. In striatum or olfactory tubercle, doses of amperozide up to 40 mg/kg did not displace radioligand binding to D2 receptors. Amperozide, haloperidol and ritanserin had similar in vivo potency in blocking the 5-HT2 binding site, but only haloperidol displaced D2 receptor binding. Based on the clinically effective dose of amperozide (0.14-0.28 mg/kg per day), it is suggested that the antipsychotic effect of amperozide is related, in part, to its in vivo interaction with the 5-HT2 receptor and that amperozide cannot be expected to exert its antipsychotic action by blockade of D2 receptors in the striatum or limbic regions.


Endogenous secretin in the rat--evidence for a role as an enterogastrone but failure to influence serum calcium homeostasis. The release of secretin into plasma by intraduodenal (id) infusion of HCl or iso-osmotic (290 mosm l-1) NaCl, and the associated changes of moderately stimulated gastric acid, serum gastrin, two calciotropic hormones, total and 45calcium (Ca), were examined in the rat. The possible role of endogenous secretin as an enterogastrone and as a mediator of the hypothesized endocrine gut-thyroid-parathyroid axis was further characterized with the aid of secretin immunoneutralization and exogenous secretion. The id-HCl-stimulated secretin, measured by a sensitive radioimmunoassay, was accompanied by a decrease in gastric acid secretion, whereas secretin blockage by anti-secretin immune serum resulted in a significant increase in acid secretion. The correlation between plasma secretin and acid output was only slight. Gastrin and Ca metabolism remained unchanged during secretin stimulation. Intravenous synthetic porcine secretin at a dose reported to be effective in other target preparations (2 CU (0.58 microgram) kg-1 h-1) had no effect on gastric acid secretion and Ca metabolism. In contrast, a pharmacological dose, 32 CU (9.3 micrograms) kg-1 h-1, inhibited acid secretion, decreased serum Ca and total protein, and increased serum parathyroid hormone, but left calcitonin and gastrin unchanged. Endogenous secretin appeared to act as an enterogastrone, but whether it was the only one is unclear. No role was detected for secretin in the gut-thyroid-parathyroid axis, since the Ca changes observed may have been unspecifically mediated.


Homologous sucrose synthase genes in barley (Hordeum vulgare) are located in chromosomes 7H (syn. 1) and 2H. Evidence for a gene translocation? The chromosomal location of the two types of sucrose synthase genes, Ss1 and Ss2, has been investigated in barley by Southern blot analysis of wheat-barley addition lines using non-cross-hybridizing-specific probes corresponding to the C-terminal regions of their respective cDNA clones (congruent to 250 bp). The Ss1 gene, whose cDNA of 2,667 bp has been entirely sequenced, is located in the beta-arm of chromosome 7H (syn. 1), while that corresponding to the homologous Ss2 is in the short arm of 2H, suggesting the existence of a translocation event between these two chromosomes in cultivated barley after an initial gene duplication and divergent evolution.


System A transport activity is stimulated in skeletal muscle in response to diabetes. We have studied the activity of system A transport in skeletal muscle during experimental diabetes. Five days after streptozotocin injection, rats showed a marked hyperglycemia and a substantial decrease in the content of GLUT-4 protein in skeletal muscle and adipose tissue. Under these conditions, basal uptake of 2-(methyl)aminoisobutyric acid (MeAIB), an index of system A transport activity, was enhanced in extensor digitorum longus (EDL) muscles from diabetic rats compared to controls. Furthermore, insulin-stimulated MeAIB uptake by the incubated EDL and soleus muscles was markedly greater in diabetic than in control rats. The derepressive phase of adaptive regulation was partially blocked in the diabetic muscle, and incubation of muscles for 3 h in the absence of amino acids led to a lower stimulation of system A transport activity in muscles from diabetic groups compared to controls. We propose that the activated system A might participate in the enhanced alanine release from muscle cells that occurs in diabetes.


Human interleukin-1 receptor antagonist. High yield expression in E. coli and examination of cysteine residues. The human IL-1 receptor antagonist (IL-1ra) was produced in a high yield E. coli expression system, and was purified in a rapid two-step purification. This recombinant IL-1ra molecule possessed full binding activity to the IL-1 receptor (type I) and totally inhibited IL-1-induced PGE2 production by human dermal fibroblasts. Radioalkylation and analysis of V8-derived IL-1ra peptides indicate that the four cysteines present in the IL-1ra are not disulphide-linked.


Changes in blood coagulation, platelet function, and plasminogen-plasmin system in diabetes. The increased risk of thromboembolism in people with diabetes mellitus is in part due to changes in the hemostatic mechanism including abnormal platelet function leading to platelet activation, increase in several coagulation factors, decrease in natural anticoagulants, and impaired fibrinolytic activity. Both microangiopathy and atherosclerosis in people with diabetes will enhance the thrombotic potential of these abnormal hemostatic changes. The recent recognition of a role of the components of the plasminogen-plasmin system in many biologic functions at the cellular level has led to studies showing that the angiopathic complications of diabetes may also be caused by impaired plasminogen activator function.


Incidence of two virulence factors (aerobactin and mucoid phenotype) among 190 clinical isolates of Klebsiella pneumoniae producing extended-spectrum beta-lactamase. Because outbreaks of multiple-resistant Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases were recently observed in French hospitals, the presence of virulence factors was examined for (i) phenotype by bioassay for aerobactin production and by culture for the mucoid phenotype, and (ii) genotype using intragenic probes of respectively 2-kb BglII and 235-bp BamHI-BglII fragments and dot-blotting among 190 unreplicated K. pneumoniae clinical isolates issued from 25 French hospitals and producing different types of extended-spectrum beta-lactamases (TEM-related enzymes: TEM-3, TEM-4, CAZ-1, CAZ-2, TEM-8, or SHV-related enzymes: SHV-2, SHV-3, SHV-4). Only 3.7% and 7% of K. pneumoniae isolates produced aerobactin and mucoid phenotypes respectively, unrelated to type of beta-lactamase. Only 2% had both factors. No discordance was reported according to the detection method tested. The low prevalence of such virulence factors seems to indicate they were not involved in dissemination of nosocomial K. pneumoniae isolates producing an extended-spectrum beta-lactamase.


Chemosensitizing effect of an antiestrogen, toremifene, on ovarian cancer. The chemosensitizing effect of an antiestrogen, toremifene, was studied on 2 human ovarian cancer cell lines in vitro and on 3 fresh surgical ovarian tumor explants with the aid of the subrenal capsule assay (SRCA). Also, 11 patients with secondarily drug resistant, recurrent gynecologic cancer (8 ovarian and 3 uterine cancers) were treated with 240 mg toremifene daily for 1 week before each course of cytostatics. Toremifene potentiated the effect of doxorubicin on both cell lines. This was also the case on 1 cell line that was not completely resistant to doxorubicin. The SRCA showed a clear potentiating effect of toremifene only on the tumor overtly resistant to the combination of cisplatin, doxorubicin, and cyclophosphamide. Of the 11 patients treated with toremifene and cytostatics, the response of 8 patients was evaluable: 3 had partial response, 3 no change, and 2 progressive disease. Toremifene seems to have a chemopotentiating effect on gynecologic drug-resistant tumors.


Pelviscopic treatment of ovarian cysts in premenopausal women. Between 1984 and 1989, 773 patients less than or equal to 45 years of age, presenting with a total of 809 ovarian cysts, underwent pelviscopy at the Department of Obstetrics and Gynecology of Kiel University. In 36 cases, cysts were bilateral. 678 cysts (84%) were treated by pelviscopy alone. Organ-preserving treatment was performed in 83%, oophorectomy or adnexectomy in only 17% of cases. Two stage Ia ovarian carcinomas (0.26% of all cysts) were operated on by pelviscopy before laparotomy. Sonography is particularly important in determining whether a pelviscopic approach is appropriate. Pelviscopic procedures are unacceptable in multilocular cysts measuring greater than or equal to 7 cm in diameter with echo-dense components. Special caution is required for any cyst measuring greater than 9 cm in diameter. The risk of opening a malignant cyst must be weighed against the advantages of pelviscopic surgery: minimal physical strain, better postoperative quality of life, and organ conservation. In doubtful cases, laparotomy is recommended.


Abnormal cardiovascular responses induced by localized high power microwave exposure. A hypothesis of microwave-induced circulatory under perfusion was tested in ketamine anesthetized rats whose heart rate, mean arterial pressure, pulse pressure, respiration rate, and body temperatures were monitored continuously. Fifty-eight ventral head and neck exposures in a waveguide consisted of sham-exposure and exposure to continuous wave (CW) and pulsed 1.25 GHz microwaves for 5 min. The 0.5 Hz (10 microseconds, 2 W average) and 16 Hz (1 microsecond, 6.4 W average) pulse-modulated microwaves were delivered at 400 kW peak power. The CW microwaves were 2 and 6.4 W. The average specific absorption rate was 4.75 W/kg per watt transmitted in the brain and 17.15 W/kg per watt transmitted in the neck. Respiration rate and mean arterial pressure were not altered. Changes in heart rate and pulse pressure were observed in rats exposed to higher power (16 Hz pulses and 6.4 W CW) but not to the lower average power microwaves (0.5 Hz pulses and 2 W CW). Depression of pulse pressure, an indication of a decrease in stroke volume, and increased (tachycardia) or decreased (bradycardia) heart rate were noted in presence of whole-body hyperthermia. The cardiac output of those animals exposed to higher average power microwaves was considered to be below normal as hypothesized. Decreased cardiac output and normal mean arterial pressure resulted in an increase in the total peripheral resistance which was contrary to the anticipated thermal response of animals.


Complement activation in the follicular light zone of human lymphoid tissues. A comparative immunohistochemical study of the distribution pattern of complement components and regulatory proteins within secondary lymphoid follicles was performed by the immunoperoxidase technique. Fifteen lymphoid tissues including appendices. Peyer's patches and tonsils were analysed. Sixty secondary lymphoid follicles with evident polarity, that is, the distinct coexistence of a light zone, dark zone and mantle zone in the same lymphoid follicle, were tested with single antibodies. The light zones were consistently immunostained in a dendritic meshwork pattern with all antibodies. The immunostaining patterns were classified into two major groups based on the immunoreactivity of the dark zone. One immunostaining pattern was characterized by no immunostaining of the dark zone to the majority of the antigens. The second group was characterized by a diffusely weak to moderate dendritic meshwork pattern of the dark zone to some of the immunostainings of C9 (monoclonal), S-protein, and DF-DRC1, and all immunostainings of CR1 (CD35), Ber-Mac-DRC (CD35), CR2 (CD21), and R4/23. All four complement regulatory proteins were localized by immunoelectron microscopy attached to the cell surface of the cells, including follicular dendritic cells, in the light zone. Our data indicate that there is an evident functional difference between the light zone and the dark zone, and that complete activation of the complement system occurs only in the light zone.


Occupancy of CD72 (the CD5 counterstructure) enhances interleukin-4-dependent CD23 expression in resting B lymphocytes. CD72, the human homologue of murine Lyb-2, was recently identified as a counterstructure to CD5. An antibody to CD72 (BU40) has been found to mimic interleukin-4 (IL-4) both in its ability to activate resting B cells into the early G1 phase of cell cycle and to augment the expression of major histocompatibility complex (MHC) class II antigen; unlike IL-4, the CD72-clustered antibody fails to induce the expression of CD23. We now report that engagement of CD72 by the IgG monoclonal antibody BU40 potentiates the capacity of IL-4--when used at optimal concentrations--to promote CD23 production in human B cells. The degree of enhancement arising from occupancy of CD72 ranged from two- to fivefold. Importantly, antibody to CD72 was also found to diminish the concentration required for IL-4 to promote CD23 expression to a level equivalent to that maximally achieved when using IL-4 alone. Engagement of CD72 by BU40 not only increased the amount of cell-associated CD23 induced by IL-4 but also led to augmented release of soluble material into the culture medium. Monovalent Fab fragments of BU40 antibody were as efficient as intact antibodies at synergizing with IL-4 for enhanced expression and release of CD23: thus simple tethering without the need for receptor cross-linking was sufficient to invoke change through CD72. Enhancement of CD23 expression via CD72 appeared to be selective for IL-4-dependent induction: the turn on of CD23 by tumour-promoting phorbol ester was left unaltered on the addition of BU40 antibody. Engagement of CD72 had no effect on the IL-4-promoted hyperexpression of surface IgM. The findings are discussed within the context of the molecular and functional interactions occurring during T-B collaboration.


Phorbol ester synergizes with Ca2+ ionophore in activation of protein kinase C (PKC)alpha and PKC beta isoenzymes in human T cells and in induction of related cellular functions. Studies described herein were designed to examine the effects of 12-O-tetradecanoyl phorbol-13-acetate (TPA), and a Ca2+ ionophore (ionomycin), singly or in combination, on the activation and expression of the Ca(2+)-dependent protein kinase C (PKC) isoenzymes (alpha, beta and gamma) at the protein and messenger RNA (mRNA) levels in T cells. These two agents induce the activation and proliferation of T lymphocytes by mimicking the action of inositol phospholipid-derived second messengers normally generated by triggering of the antigen-specific T-cell receptor (TcR)/CD3 complex. TPA-induced T-cell proliferation, expression of interleukin-2 receptor-alpha subunit (IL-2R alpha) and transferrin receptor, CD3 down-regulation and, lastly, the cytosol-to-membrane PKC translocation (determined by an enzymatic assay or by immunoblotting with a cross-reactive anti-PKC peptide antibody) were all facilitated by ionomycin. Immunoblots with isoenzyme-specific anti-PKC monoclonal antibodies demonstrated expression of immunoreactive PKC alpha, PKC beta and PKC gamma proteins that were translocated to the membrane upon TPA plus ionomycin stimulation. Resting T cells expressed abundant levels of mRNA for PKC alpha and PKC beta, but very low levels (relative to brain) of PKC gamma. TPA increased by two- to threefold the expression of PKC beta, but not of PKC alpha or PKC gamma, mRNA within 12 hr of stimulation. Ionomycin synergized with TPA in increasing the expression of PKC alpha and PKC beta mRNA. The two agents also synergized in inducing expression of additional activation/growth-associated genes, namely the c-myc protooncogene, ornithine decarboxylase (ODC) and IL-2R alpha. Ionomycin alone was inactive (or marginally active) in all of these assays. The translocation of distinct Ca(2+)-dependent PKC isoenzymes to the membrane and the up-regulation of PKC alpha and beta mRNA suggest that at least these two isoenzymes are involved in discrete steps of the pathway leading to T-cell activation and proliferation. Moreover, the combined effects of TPA and ionomycin on T-cell function and cell-surface antigen expression appear to be due, at least in part, to their synergistic activation of distinct PKC isoenzyme(s).


Altered patterns of glycosylation on rat lymphocytes associated with activation. The expression of cell-surface carbohydrates on rat lymphocytes was investigated by flow cytometry using a panel of lectins. A small group of lectins was identified, all with a main binding requirement of N-acetylgalactosamine that bound to all B lymphocytes but only to activated T lymphocytes expressing the interleukin-2 (IL-2) receptor (as shown by staining with the monoclonal antibody OX39). Studies demonstrated that five of these lectins competed for the same binding site, while others did not. With the knowledge of the binding requirements of these lectins, a structure can be deduced for the carbohydrate moiety which appears on T lymphocytes when activated.


Interleukin-5 increases the expression of alkaline phosphatase activity in murine B lymphocytes. Interleukin-5 (IL-5) was shown to enhance, in a dose-dependent fashion, the expression of alkaline phosphatase (APase) activity in splenic B cells stimulated with dextran sulphate (DXS). The potentiating effect of IL-5 was still more evident when assayed in large B cells than in small resting B cells, whereas IL-2, IL-4 and IL-6 were devoid of activity. Concomitant with increased APase expression, cell-cycle analysis by flow cytometry showed that large B cells in the early G1 phase were stimulated by IL-5, in conjunction with DXS, to enter G1B and to progress further through S and G2/M. A phosphorylation-dephosphorylation pathway could, thus, be involved in IL-5 transmembrane signalling.


Expression of mouse Tla region class I genes in tissues enriched for gamma delta cells. The Tla region of the BALB/c mouse major histocompatibility complex contains at least 20 class I genes. The function of the products of these genes is unknown, but recent evidence demonstrates that some Tla region gene products could be involved in presentation of antigens to gamma delta T cells. We have generated a set of polymerase chain reaction (PCR) oligonucleotide primers and hybridization probes that permit us to specifically amplify and detect expression of 11 of the 20 BALB/c Tla region genes. cDNA prepared from 12 adult and fetal tissues and from seven cell lines was analyzed. In some cases, northern blot analysis or staining with monoclonal antibodies specific for the Tla-encoded thymus leukemia (TL) antigen were used to confirm the expression pattern of several of the genes as determined by PCR. Some Tla region genes, such as T24d and the members of the T10d/T22d gene pair, are expressed in a wide variety of tissues in a manner similar to the class I transplantation antigens. The members of the TL antigen encoding gene pair, T3d/T18d, are expressed in only a limited number of organs, including several sites enriched for gamma delta T cells. Other Tla region genes, including T1d, T2d, T16d, and T17d, are transcriptionally silent and transcripts from the T8d/T20d gene pair do not undergo proper splicing. In general, sites that contain gamma delta T lymphocytes have Tla region transcripts. The newly identified pattern of expression of the genes analyzed in sites containing gamma delta T cells further extends the list of potential candidates for antigen presentation to gamma delta T cells.


Influence of cefodizime on the reagibility of human leukocytes. Cefodizime was evaluated for its effect on a number of parameters of leukocyte function in humans. Four healthy volunteers received 2 g i.v. b.i.d. for seven days. Leukocyte activity was measured before, during and after treatment. Using opsonized zymosan as a stimulant, no effect on the respiratory burst of granulocytes was observed. It was found, however, that the lymphocytes in three of the four subjects showed significantly more marked proliferation rates in the mixed lymphocyte reaction after administration of cefodizime than at baseline. The stimulation indices subsequently returned to normal. This pilot study therefore demonstrated that cefodizime has biological response modifying properties in healthy humans.


Identification of childhood disability in Jamaica: the ten question screen. This is the first in a series of papers that report the testing of two instruments for the identification and assessment of childhood disability by community workers (CWs) in Third World countries. It is part of the International Epidemiologic Study on Childhood Disability. The Ten Question Screen (TQ) was used as the main instrument to identify disability in a two stage population-based survey of 5478 children aged 2-9 years in Clarendon, Jamaica. In the second stage, TQ positive and 8% of the screen negative controls were professionally assessed by a doctor and a psychologist using standard criteria based on the main classification system of the ICIDH. Sensitivity of the TQ as a whole varied in different strata of the group and amongst different disabilities, from perfect in girls under 6 years, fits and motor disabilities and for serious disability in all group except boys over 5 years with cognitive disability. Specificity was good but the false positive rate was unacceptably high at 74%. It was concluded, firstly, that the validation of a simple questionnaire of perceptions of behaviour against objective measurements of impairments was perhaps not fair to the TQ. In spite of this, the TQ would be a very useful instrument in collecting disability data or for identifying people in need of rehabilitation help, if a way of reducing false positives could be found.


The cancer-associated retinopathy antigen is a recoverin-like protein. Cancer-associated retinopathy (CAR) is a rare form of retinal degeneration that occurs in association with certain forms of cancer. CAR patients typically possess high titers of autoantibodies against a specific photoreceptor protein--the 23 kD retinal CAR antigen. The mechanisms involved in the vision loss experienced by CAR patients are not understood, but serologic studies indicate the process could include a series of autoimmune reactions directed at specific components of the retina. Because the retinal CAR antigen is the principal ocular autoantigen involved in the antibody response of CAR patients, characterizing it would contribute to the understanding of putative autoimmune involvement. Serum antibodies from CAR patients have been used to isolate the gene encoding the CAR antigen from a cDNA library of human retina. Nucleotide sequence analysis suggests that the CAR antigen shows approximately 90% homology to the published amino acid sequence of bovine recoverin.


Alterations in ascorbic acid transport into the lens of streptozotocin-induced diabetic rats and guinea pigs. High ascorbic acid (AA) levels in the aqueous humor and intraocular tissues, including the lens, are thought to protect against the harmful effects of photochemical and ambient oxidation reactions involving oxygen and its radicals. In addition, AA may have various metabolic functions, including structural collagen formation in intraocular tissues. Recent work showed that, in the guinea pig, reduced AA was concentrated in the aqueous and lens epithelium. These in vivo studies were extended to streptozotocin-induced diabetic rats and guinea pigs to explore the state of AA transport and passive L-glucose movement in the diabetic lens. A bolus dose of radiolabeled test molecules, including 14C-AA, 3H-L-glucose (L-glu), and 14C-3-O-methyl-D-glucose, was injected into the blood at time zero, and the time-dependent concentrations of these labeled molecules were determined as they move into the aqueous humor, lens epithelium and capsule, and interior compartments. These kinetic studies provided a unique measurement of the functioning state of passive and carrier transport mechanisms in situ in normal and diabetic animals. Diabetic animals (blood glucose, greater than 300 mg/dl) were categorized in terms of the length of time of uniform monitored drug-induced diabetes as short term (10-20 days); midterm (40-60 days), and long term (100+ days). In the rat lens epithelium, significant decrease occurred in the active movement of AA (control KEi, 0.693 +/- 0.062 [n = 12]; midterm drug-induced diabetes Ki, 0.192 +/- 0.054 [n = 10]; t-test P less than 0.001). The passive L-glu entry rate increased (control KEi, 0.0268 +/- 0.0053 [n = 12]; midterm drug-induced diabetes KEi, 0.0421 +/- 0.075 [n = 10]; t-test P less than 0.005). Thus, it was suggested that the drug-induced diabetic rat lens epithelium had lost some of its ability to concentrate AA to high levels and achieved epithelial levels only one- to twofold those of aqueous; control animals concentrated AA to levels of five- to eightfold those of aqueous within 20 min. By contrast, the rate of movement of L-glu from epithelium to stroma increased minimally (control KSi, 0.0116 +/- 0.021 [n = 12]; midterm drug-induced diabetes KSi, 0.0136 +/- 0.034 [n = 10]; t-test P less than 0.05). In addition, AA entry rate into lens cortex increased fourfold (control KSi, = 0.0018 +/- 0.0003 [n = 12]; midterm drug-induced diabetes KSi, 0.0081 +/- 0.024 [n = 10]; t-test P less than 0.001).(ABSTRACT TRUNCATED AT 400 WORDS)


The transformation-defective adenovirus 12 host range mutant CS-1 lacks the E1a-specific 9.5S mRNA and contains a second deletion in E1b. We have further analyzed structure and expression of the E1 region of the transformation-defective adenovirus 12 (Ad12) host range mutant CS-1. Using cDNA polymerase chain reaction analysis, the E1a region was found to give raise to five transcripts analogous to the Ad12wt 13S, 12S, 11S, 10S species and the normal 9S mRNA. Due to loss of a splice acceptor site at position 852, the Ad12wt-specific 9.5S transcript cannot be synthesized by the mutant CS-1. The fact that the virus is, however, completely viable indicates that this mRNA is dispensable for lytic growth of Ad12. Besides the deletions in E1a described previously, a second deletion of 31 base pairs was found in the E1b gene. It affects the start site of the E1b-specific mRNAs and destroys or eliminates the AUG sequence for the E1b 19-kD protein. As shown earlier, normal E1b mRNA expression is found in mutant-infected cells, but no 19-kD E1b protein was immunoprecipitated by an E1-specific rat tumor serum.


Mycotoxins and reproduction in domestic livestock. Molds are parasitic plants that are ubiquitous in livestock feedstuffs. Even though molds themselves reduce the quality of grains, their synthesis of chemical substances termed mycotoxins causes the greatest monetary loss to the animal industry. Five major mycotoxins that impair growth and reproductive efficiency in North America are aflatoxins, zearalenone, deoxynivalenol, ochratoxin, and ergot. Aflatoxins are produced by Aspergillus flavus and Aspergillus parasiticus. Consumption of grains containing aflatoxins by swine affects reproduction indirectly by reducing feed intake and growth. In swine, aflatoxins impair liver and kidney function, delay blood clotting, increase susceptibility to bruising, and interfere with cellular humoral immune systems. Ruminants are comparatively resistant to aflatoxicosis, but presence of aflatoxins in milk of dairy cows is closely monitored for human safety. Depending on environmental conditions, Fusarium roseum can produce either zearalenone or deoxynivalenol. Days 7 to 10 postmating seem to be a critical period of gestation for zearalenone to exert its detrimental actions on early embryonic development. Presence of deoxynivalenol in swine feedstuffs decreases feed intake, causes feed refusal, and induces occasional vomiting. Several species of Penicillium and Aspergillus produce ochratoxin, a mycotoxin that causes necrosis of kidney tissue. Ergot alkaloids produced by Claviceps purpurea on wheat can cause reproductive problems and are associated with lactational failure in swine. Various methods have been developed to remove mycotoxins from infected feedstuffs. Chemical analyses in laboratories as well as diagnostic kits suitable for use at the elevator or farm can be used successfully to identify which mycotoxins are present in suspect feedstuffs.


Rat soleus muscle fiber responses to 14 days of spaceflight and hindlimb suspension. Morphological and enzymatic responses in fibers expressing fast, slow, or both types of myosin heavy chain (MHC) were studied in rats after 14 days of spaceflight (COSMOS 2044) or hindlimb suspension. Although the percentage of slow-twitch fibers was unchanged, a higher percentage of fibers that expressed both slow and fast MHC was observed in flight and suspended rats than in synchronous ground-based controls. The soleus was 25 and 34% smaller than control after 14 days of flight and suspension, with the reduction in fiber cross-sectional area (CSA) being greater in slow- than in fast-twitch fibers in both experimental groups. The activities of succinate dehydrogenase (SDH) and alpha-glycerophosphate dehydrogenase (GPD) were not significantly affected by flight or suspension. The total SDH activity (i.e., SDH activity x CSA) decreased significantly in the slow-twitch fibers of the flight and the fast-twitch fibers of the suspended rats, in large part due to fiber atrophy. A shift in MHC expression in 14 and 9% of the fibers in flight and suspended rats occurred without a change in myosin adenosinetriphosphatase activity. The SDH and GPD activities of the fibers that expressed both slow and fast MHC were slightly higher than the slow-twitch fibers and slightly lower than the fast-twitch fibers. These data indicate that events were initiated within 14 days of spaceflight or suspension that began to reconfigure the protein profiles of 9-14% of the slow-twitch fibers from typical slow-twitch toward those of fast-twitch fibers, while all fibers were dramatically losing total protein.(ABSTRACT TRUNCATED AT 250 WORDS)


Adaptation of fibers in fast-twitch muscles of rats to spaceflight and hindlimb suspension. The adaptation of single fibers in medial gastrocnemius (MG), a fast-twitch extensor, and tibialis anterior (TA), a fast-twitch flexor, was studied after 14 days of spaceflight (COSMOS 2044) or hindlimb suspension. Cross-sectional area (CSA) and succinate dehydrogenase (SDH), alpha-glycerophosphate dehydrogenase (GPD), and myofibrillar adenosinetriphosphatase (ATPase) activities were determined in fibers identified in frozen serial cross sections. Fibers were categorized as light, dark, or intermediate on the basis of myosin ATPase staining and alkaline preincubation and immunohistochemically as reacting with slow, fast, or both slow and fast myosin heavy chain monoclonal antibodies. Because there was a close relationship between these two means of categorizing fibers, all fibers were categorized on the basis of the immunohistochemical reaction. The percentage of slow- and fast-twitch fibers of the MG and TA were unchanged in either group. Mean fiber size of all fibers, irrespective of type, was unaffected in either muscle after flight or suspension. The fibers that expressed both fast and slow myosin heavy chains were smaller than control in the MG of both experimental groups. Compared with control, the SDH and total SDH activities in the MG were significantly less in suspended rats, with the fast-twitch fibers showing the largest difference. The ATPase activity in the MG was higher in flight than in control or suspended rats. There were no significant effects of flight on fibers of the TA. In contrast, the TA in suspended rats had higher GPD activities than either control or flight rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of microgravity and tail suspension on enzymes of individual soleus and tibialis anterior fibers. Selected enzymes of energy metabolism were measured in random individual fibers of soleus and tibialis anterior (TA) muscles from rats exposed for 2 wk to spaceflight (F) aboard COSMOS 2044 or tail suspension (T) and from synchronous controls. Average size of soleus fibers (dry weight per unit length) was reduced 37% in F and T fibers; there was little change in TA fibers. Enzyme changes were more pronounced in soleus than in TA fibers. Three enzymes characteristic of fast-twitch muscles, pyruvate kinase, glycerol-3-phosphate dehydrogenase, and 1-phosphofructokinase, were elevated in F and T soleus fibers, but changes in phosphofructokinase were not statistically significant. 3-Ketoacid-CoA transferase, characteristic of slow-twitch muscles, did not change significantly in either F or T fibers. Hexokinase, usually moderately higher in slow- than in fast-twitch muscles, increased markedly in both F and T fibers. In TA fibers analyzed for hexokinase, malate dehydrogenase, phosphohexoisomerase, and pyruvate kinase, only hexokinase and malate dehydrogenase showed significant changes. Hexokinase increased 83% in one of two T muscles. Enzyme data for TA fibers typed by myosin adenosinetriphosphatase were more informative: phosphofructokinase, phosphorylase, and glycerol-3-phosphate dehydrogenase were increased in type IIb fibers of either F or T muscles or both. Malate dehydrogenase was not changed in fibers of any type in either F or T muscle.


Alpha 6.beta 1 integrin (laminin receptor) is down-regulated by tumor necrosis factor alpha and interleukin-1 beta in human endothelial cells. Endothelial cells from human umbilical vein (HEC) express several distinct integrin complexes that mediate the interaction with the basal membrane components. In this paper we show that treatment with tumor necrosis factor alpha (TNF alpha) down-regulates the expression of the laminin receptor alpha 6.beta 1 integrin in cultured HEC. After 48 h of treatment with TNF alpha, the level of expression of the alpha 6.beta 1 complex reached 20% of the control value. The down-regulation of the alpha 6.beta 1 integrin is caused by a decreased expression of the alpha 6 subunit, whereas the synthesis of the beta 1 subunit remains constant. Northern blot analysis shows that the decreased level of alpha 6 subunit synthesis is caused by down-regulation of alpha 6 mRNA in TNF alpha-treated HEC. TNF alpha treatment does not alter the expression of alpha 2, alpha 3, and alpha 5 integrins, also present on endothelial cell surface, thus showing that this cytokine has a selective action on distinct integrin complexes. Down-regulation of alpha 6.beta 1 correlates with pronounced reduction in adhesion of TNF alpha-treated HEC to laminin, but not to fibronectin-coated culture dishes. In addition to TNF alpha, interleukin-1 beta also decreases the expression of the alpha 6.beta 1 integrin and reduces adhesion to laminin, thus suggesting that this regulation plays an important role in inflammation.


Allosteric modulation of Leishmania donovani plasma membrane Ca(2+)-ATPase by endogenous calmodulin. The plasma membrane of the human pathogen Leishmania donovani possesses a high-affinity transmembrane Ca(2+)-ATPase that has its catalytic site oriented toward the cytoplasmic milieu (Ghosh, J., Ray, M., Sarkar, S., and Bhaduri, A. (1990) J. Biol. Chem. 265, 11345-11351). When the enzyme is studied in its more authentic, physiologically relevant, membrane-associated form, it exhibits pronounced sigmoidal kinetics with Ca2+ (K0.5 approximately 700 nM) in a trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid buffering system that effectively complexes all available Mg2+. Addition of exogenous Mg2+ (60 microM) completely abolishes sigmoidicity and establishes strictly hyperbolic kinetics, and the Km for Ca2+ reduces to 100 nM. Mg2+ can be replaced by heterologous calmodulin. The exclusive dependence of the enzyme on only Ca2+ for its activity and its positive allosteric modulation by Mg2+ distinguish this enzyme from other well-characterized plasma membrane Ca(2+)-ATPases. Employing this Ca(2+)-ATPase as the assay system, a soluble endogenous activating protein factor was purified that, by several criteria, corresponds to authentic calmodulin. The parasite calmodulin shifts the kinetics to hyperbolic kinetics, increases the Vmax 2-fold, and most important lowers the Km (approximately 100 nM) to a physiological level. The interaction with endogenous calmodulin thus converts the enzyme from a totally inactive to a fully active state.


Action of neopullulanase. Neopullulanase catalyzes both hydrolysis and transglycosylation at alpha-(1----4)- and alpha-(1----6)-glucosidic linkages. The transglycosylation reaction catalyzed by neopullulanase was analyzed. Radioactive oligosaccharides were produced when the enzyme acted on maltotriose in the presence of [U-14C]glucose. Some of the radioactive oligosaccharides had only alpha-(1----4)-glucosidic linkages, but others were suggested to have alpha-(1----6)-glucosidic linkages. The existence of alpha-(1----6)-glucosidic linkages in the products from maltotriose with neopullulanase was proven by proton NMR spectroscopy and methylation analysis. We previously reported that the one active center of neopullulanase catalyzes the hydrolysis of alpha-(1----4)- and alpha-(1----6)-glucosidic linkages (Kuriki, T., Takata, H., Okada, S., and Imanaka, T. (1991) J. Bacteriol. 173,6147-6152). These facts proved that neopullulanase catalyzed all four types of reactions: hydrolysis of alpha-(1----4)-glucosidic linkage, hydrolysis of alpha-(1----6)-glucosidic linkage, transglycosylation to form alpha-(1----4)-glucosidic linkage, and transglycosylation to form alpha-(1----6)-glucosidic linkage. The four reactions are typically catalyzed by alpha-amylase, pullulanase, cyclomaltodextrin glucanotransferase, and 1,4-alpha-D-glucan branching enzyme, respectively. These four enzymes have some structural similarities to one other, but reactions catalyzed by the enzymes are considered to be distinctive: the four reactions are individually catalyzed by each of the enzymes. The experimental results obtained from the analysis of the reaction of the neopullulanase exhibited that the four reactions can be catalyzed in the same mechanism.


The kinetics for the phosphoryl transfer steps of the sarcoplasmic reticulum calcium ATPase are the same with strontium and with calcium bound to the transport sites. Rate constants for most of the steps of the reaction cycle of the sarcoplasmic reticulum calcium-ATPase are similar or identical with Ca2+ or Sr2+ as the transported ions in spite of the large differences in the size and affinity of Ca2+ and Sr2+ (5 mM MgCl2, 100 mM KCl, pH 7.0, 25 degrees C). Phosphorylation of cE.Sr2 and cE.Ca2 by ATP occurs with kp = 220-235 s-1, whereas phosphorylation of E.ATP+Ca2+ or Sr2+ is consistent with kb = 50-70 s-1. Hydrolysis of E approximately P.Sr2 and E approximately P.Ca2 occurs with kt = 20 s-1, and the addition of 7 mM ADP to E approximately P.Sr2 or to E approximately P.Ca2 gives a burst of approximately 43% dephosphorylation, followed by dephosphorylation with k = 46 s-1. However, one Sr2+ ion dissociates from cE.Sr2 and from cE.ATP.Sr2 with k congruent to 120 s-1, whereas one Ca2+ ion dissociates from cE.Ca2 with k = 38 s-1 and from cE.ATP.Ca2 with k = 80 s-1.