Predictive value of total body bone mineral density for vertebral fractures in elderly women. Geriatric Study Group.

In a general population-based geriatric disease survey in Taipei City, the bone mineral density (BMD) of 58 women over 65 years of age was measured for the whole body, lumbar spine (L2-L4), and proximal femurs using a 153Gd based dual photon absorptiometer. These women were found to have at least one vertebral fracture. The results showed that the BMD readings of both the lumbar spine (L2-L4) (mean Z score +/- SEM = 0.05 +/- 0.12) and the femoral neck (mean Z score +/- SEM = -0.20 +/- 0.10) were not statistically different from those of age-matched controls. However, the total body BMD in these 58 patients was significantly lower than in the normal controls (mean Z score +/- SEM = -1.07 +/- 0.10, p < 0.0001). In the normal control group (N = 69, age 50-85), there was a significant linear correlation between the total body and lumbar BMD (r = 0.81, p < 0.0001). This correlation was not found in the 58 women with vertebral fractures (r = 0.14, NS). Our results suggest that geriatric women with vertebral fractures are more osteoporotic than normal aged women, even though they have a relatively mild degree of spinal osteoporosis. But, because of age-associated degenerative changes or other factors, conventional anteroposterior lumbar BMD measurements cannot detect the difference. The total body BMD readings, but not the lumbar or femoral neck BMD readings, seem to be less affected by these local changes and may provide a better discriminative or predictive value for vertebral fracture in this particular age group.


Management of infected tibial intramedullary nailing using an organized treatment protocol.

Twenty cases of osteomyelitis, following intramedullary nailing of tibial shaft fractures, were managed with a prospective treatment protocol, comprising intramedullary reaming debridement, antibiotic-bead depot, external skeletal fixation, microvascular muscle flap and early cancellous bone grafting. The follow-up period ranged from 25 to 48 months (average, 34.3 months). Pseudomonas aeruginosa (37.5%) and Staphylococcus aureus (20.8%) were the organisms most commonly involved. There were eight united and 12 ununited fractures after reaming debridement surgery. Nineteen infections were initially arrested by one debridement. One infection was arrested by two Sequential debridements. All 12 ununited fractures were stabilized by Hoffmann unilateral external fixation until the fracture healed. The time spent in external fixation ranged from three to seven months (average, 5.2 months). Early cancellous bone grafting was successfully accomplished for nine ununited fractures with major debridement bone loss. The average union time of the nine fractures with bone grafting was 6.5 months (range, from six to eight months). We believe that this treatment protocol gives a predictable and rapid recovery. The complications were infection recurrence in two cases at the old tibial shaft fracture sites, minor Hoffmann pin tract infection in two cases and stiffness in two ankles and one knee.


Experimental structural scoliosis in rabbits.

Structural scoliosis was produced by purely posterolateral mechanical tethering of the spine with ligation of the scapula to the opposite pelvic bone in rabbits during the growing period. It is a simple and easy method without direct trauma to the spine. When the tethering force occurred discontinuously or only in the coronal plane by drawing of the scapula to the ipsilateral pelvic bone in 17 rabbits, spinal deformity could not be produced. The model provided evidence of biplanar deformity as one of the initial factors for the development of structural scoliosis.


Dissociation of inhibitory effects of low-dose ASA on thromboxane production and platelet aggregation in ischemic stroke patients.

Acetylsalicylic acid (ASA) inhibits thromboxane production and hence platelet aggregation. However, individual variations in platelet aggregability and serum thromboxane B2 (TxB2) concentration after a low dose of ASA (40 mg/day) have been reported. To clarify this issue, we studied plasma thromboxane levels and platelet aggregation in 43 ischemic stroke patients. Of the 22 patients who received 100 mg of ASA daily, dissociation between inhibitory effects of ASA on the plasma TxB2 level and threshold concentrations of adenosine diphosphate was found in three cases after one month of drug administration, and in three cases after six, 12 and 18 months of ASA therapy. This dissociation also developed in two patients after one month and six months, respectively, of treatment in the 21 patients who received 300 mg of ASA daily. The dissociation between the inhibitory effects on plasma TxB2 and the circulating platelet aggregate ratio was found in two cases after taking medication for one month, and in four cases after six, 12, 18 and 24 months of therapy in the 100 mg ASA group. In the 300 mg ASA group, dissociation was noted in two cases after one month of medication, and in two cases after six and 12 months of medication. In these patients, although their TxB2 levels were inhibited to almost unmeasurable levels, platelet aggregation was still not inhibited. This ASA inhibitory dissociation phenomenon on platelet function may be due to the low dose of ASA, individual differences in platelet function in response to ASA therapy, or factors other than those involved in the cyclooxygenase system.


Intracavernous self-injection therapy for the treatment of erectile dysfunction.

Fifty-one patients with chronic erectile dysfunction were selected for this study involving intracavernous self-injection therapy with papaverine or prostaglandin E1 (PGE1). Patients were screened and grouped as follows: pronounced vasculogenic, mild vasculogenic, venous leakage, neurogenic and psychogenic impotence. The duration of follow-up in these 51 patients was from 1.5-30.5 months (average, 11.8 months). The average effective dosage of papaverine and PGE1 was variable among the different groups. This kind of therapy proved to be effective in our preliminary results, which showed that 35 patients (68.6%) were found to be good responders and eight patients (15.7%) were temporary responders. In our detailed questionnaire, answered by all 51 patients, we found 29 patients (56.9%) had increased their frequency of sexual activity, 38 patients (74.5%) had sustained their erection more than they ever had before, and 43 patients (84.3%) had enjoyed sexual orgasm following this pharmacologically assisted erection. Compared with papaverine, PGE1 was found to have fewer complications. None of the patients complained of any discomfort after long-term, self-injection with PGE1. However, two patients (3.9%) had sustained erections and two patients (3.9%) developed palpable fibrotic plaque after papaverine injection in our study.


Extracorporeal shock-wave lithotripsy of gallbladder stones.

Recently, extracorporeal shock-wave lithotripsy has been introduced as a nonoperative treatment for gallstone disease. To evaluate its applicability in Taiwan, where cholesterol stones are not very frequent, 18 patients out of 194 (9.3%) were selected according to the Munich inclusion criteria to receive this treatment. Chenodeoxycholic acid and ursodeoxycholic acid were administered as adjunctive litholytic therapy. Twenty-eight sessions of lithotripsy were performed, resulting in a 83% satisfactory fragmentation rate (< or = 5 mm). No serious complications were encountered; however, two patients later underwent laparoscopic cholecystectomy. The stones disappeared in four patients after treatment, but recurred in one after discontinuation of oral bile acids. The long-term result was discouraging compared to European reports. A 13% stone-free rate was noticed at one year and 25% was anticipated during the 13-18 month follow-up. It is our conclusion that low eligibility, low cost-effectiveness and poor treatment results will restrict the wide-spread use of this nonoperative therapy in Taiwan.


Malignant rhabdoid tumor arising from soft parts of the right thigh with unusual neurologic manifestation: report of a case.

A case of malignant rhabdoid tumor (MRT) arising from the soft tissue of the right thigh in a 49-year-old Chinese female with peripheral neuropathy is reported. The tumor, exhibiting the salient features of MRT, was composed of sheets and nests of polygonal cells with prominent nucleoli and characteristic paranuclear inclusion-like hyaline globules under light microscopy which corresponded to aggregates of intermediate filaments under electron microscopy. The results of immunohistochemical studies of the tumor cells were also characteristic: cytokeratin (+), vimentin (+), epithelial membrane antigen (EMA) (+), desmin (-), myoglobin (-), leukocyte common antigen (LCA) (-), kappa (-), lambda (-), IgG (-) and IgA (-). Serologic study revealed an M-component of IgA. The clinical evolution of the patient was highly aggressive and inevitably lethal. An adult malignant rhabdoid tumor is unusual, and its association with peripheral neuropathy and the coexistence of an M-component of IgA in this case appears to be unique. In this report, the differential diagnosis of histopathologic features, the association of peripheral neuropathy and the coexistence of an M-component of IgA are discussed.


Diffuse panbronchiolitis: report of a case.

A 33-year-old male presented with a productive cough of yellowish sputum which he had had for several years and progressive dyspnea on exertion that had been present for one year. Physical examination on admission disclosed clubbing of the fingers, diffuse inspiratory crackles and some rhonchi on auscultation. Plain chest film showed diffuse fine nodular lesions in both lungs. Pulmonary function tests demonstrated obstructive ventilatory impairment with a positive bronchodilator response. A CT scan of the chest showed diffuse fine nodular infiltrations in both lung fields. Arterial blood gas analysis of the patient, while breathing room air, revealed mild hypoxemia. The histologic findings of an open lung biopsy specimen were compatible with a diagnosis of diffuse panbronchiolitis. The patient was treated with erythromycin and a bronchodilator, and was regularly followed at the outpatient department. In this report, clinical manifestations, diagnostic criteria and recent advances in the treatment of diffuse panbronchiolitis are discussed.


Hyperoxaluria, nephrolithiasis, nephrocalcinosis and renal failure after massive resection of the small intestine: report of a case.

Hyperoxaluria is frequently seen in patients with inflammatory bowel disease, or after resection of the ileum. It is assumed to be responsible for the development of nephrolithiasis, nephrocalcinosis (oxalate nephrosis) and progressive renal impairment in these patients. Steatorrhea may aggravate the severity of hyperoxaluria. A 60-year-old male underwent massive resection of the jejunum and ileum 10 years prior to admission, due to strangulation of the small bowel, with occlusion of the superior mesenteric artery. He remained well except for steatorrhea which developed two-and-a-half years prior to admission, when microhematuria, proteinuria and oxaluria developed progressively. Since that time, the nephrolithiasis, nephrocalcinosis and renal failure have continued to worsen despite therapy with oxalate restriction and oxalate-binding agents. A renal biopsy, performed late in the clinical course, showed severe changes in the renal parenchyma. The decline in renal function proved irreversible. The unusual metabolic consequences of massive resection of the small intestine and their mechanisms are discussed.


Choriocarcinoma presenting as a unilateral renal mass and gross hematuria in a male: report of a case.

Choriocarcinoma of the kidney, either primary or metastatic, is rarely reported in the literature. We encountered a male patient with choriocarcinoma of the right kidney. The patient initially presented with fever, gross hematuria and flank pain. He had initially been unsuccessfully treated for bilateral renal stones for four months before an accurate diagnosis was made. The disease was verified only after a resection of the right kidney and meticulous examination of microscopic sections of the tumor. Further immunohistochemical study strongly supported the diagnosis of choriocarcinoma. The tumor responded briefly to Cisplatinum, Vinblastine and Bleomycin (PVB) treatment. The patient died of brain metastasis and respiratory failure four months after starting systemic chemotherapy.


Single lung transplantation for end-stage silicosis: report of a case. Lung Transplant Group.

After successful animal studies since 1986, a single left lung transplantation was performed on a 35-year-old male patient with end-stage silicosis in July 1991. The surgical technique was similar to that used by the Toronto Transplant Group, except for some modification in bronchial anastomosis. The donor lung was preserved by simple surface cooling after the administration of heparin, methylprednisolone and PGE1. The ischemic time for the donor lung was 3.5 hours. A cardiopulmonary bypass through the femoral vessels was applied for a duration of 90 minutes. Immediate postoperative complications included massive bleeding, disseminated intravascular coagulation, adult respiratory distress syndrome and acute graft rejection. Fortunately, we overcame these complications through intensive care. Immunosuppression included antilymphocytic globulin, cyclosporine, azathioprine and corticosteroids. The results of this single lung transplantation were satisfactory. The patient was doing well and was able to satisfactorily breathe room air six weeks after the transplantation. Unfortunately, the patient died of opportunistic systemic aspergillosis six months after the transplantation. In conclusion, lung transplantation is an effective treatment for patients with end-stage lung diseases, and the results of this first single lung transplantation in Taiwan are encouraging and promising.


Localization in situ of polyhomeotic transcripts in Drosophila embryos reveals spatially restricted expression beginning at the blastoderm stage.

Polyhomeotic is a member of a group of genes, the Pc-group responsible for the maintenance of gene expression during development. In particular, the Pc-group of genes is involved in the correct expression of homeotic genes of the bithorax and Antennapedia complexes. Molecular analysis reveals that the Pc-group genes function relatively late in development, once homeotic gene expression has been correctly initiated. This initiation of homeotic gene expression depends on interaction between genes in the segmentation gene hierarchy, the gap and pair-rule genes. The in situ analysis presented here indicates that polyhomeotic transcripts are expressed in a spatially restricted fashion early in development. This blastoderm expression is under the control of genes in the segmentation hierarchy. Given these results, and the role of polyhomeotic in the correct maintenance of homeotic gene expression, I propose that polyhomeotic expression may relay the initiation of homeotic gene expression with other mechanisms involved in the maintenance of this expression, involving the other Pc-group genes.


Cloning and sequence comparison of the mouse, human, and chicken engrailed genes reveal potential functional domains and regulatory regions.

We have isolated and characterized genomic DNA clones for the human and chicken homologues of the mouse En-1 and En-2 genes and determined the genomic structure and predicted protein sequences of both En genes in all three species. Comparison of these vertebrate En sequences with the Xenopus En-2 [Hemmati-Brivanlou et al., 1991) and invertebrate engrailed-like genes showed that the two previously identified highly conserved regions within the En protein ]reviewed in Joyner and Hanks, 1991] can be divided into five distinct subregions, designated EH1 to EH5. Sequences 5' and 3' to the predicted coding regions of the vertebrate En genes were also analyzed in an attempt to identify cis-acting DNA sequences important for the regulation of En gene expression. Considerable sequence similarity was found between the mouse and human homologues both within the putative 5' and 3' untranslated as well as 5' flanking regions. Between the mouse and Xenopus En-2 genes, shorter stretches of sequence similarity were found within the 3' untranslated region. The 5' untranslated regions of the mouse, chicken and Xenopus En-2 genes, however, showed no similarly conserved stretches. In a preliminary analysis of the expression pattern of the human En genes, En-2 protein and RNA were detected in the embryonic and adult cerebellum respectively and not in other tissues tested. These patterns are analogous to those seen in other vertebrates. Taken together these results further strengthen the suggestion that En gene function and regulation has been conserved throughout vertebrate evolution and, along with the five highly conserved regions within the En protein, raise an interesting question about the presence of conserved genetic pathways.


Mutations in the glutamine synthetase I (gsI) gene produce embryo-lethal female sterility in Drosophila melanogaster.

A female-sterile mutation (fs(2) PM11-19) was recovered in a screen for P-M hybrid dysgenesis induced mutations uncovered by a deletion of region 21B and was identified as an allele of the gene encoding the Drosophila glutamine synthetase I (GSI) mitochondrial isozyme. Molecular analysis has shown that fs(2)PM11-19 contains a 5 kb insert within 500 bp upstream of the transcriptional start site of the gsI gene. Mutant flies have extremely low levels of gsI transcription and GSI activity. A pre-existing deficiency (Df(2L) netPM1) with a breakpoint near the transcription start site was also found to be a female-sterile allele of gsI. All eggs laid by PM11-19 homozygous females, as well as by females heterozygous for this mutation and a deletion or any of several recessive lethal alleles of the gsI gene, fail to hatch. We conclude that an adequate level of maternally supplied GSI activity is necessary in the early stages of Drosophila embryonic development.


Automated measurement of lactate dehydrogenase, alkaline phosphatase and gamma-glutamyltransferase in urines: an alternative to the manual procedure.

A sensitive and precise automated assay of urinary lactate dehydrogenase (EC 1.1.1.27), alkaline phosphatase (EC 3.1.3.1) and gamma-glutamyltransferase (EC 2.3.2.2) is described. For this purpose, we used a BM/Hitachi System 704 model and reagents for automated analysis of serum enzymes from Boehringer Mannheim. However, the schedules of enzyme chemistry parameters recorded by the autoanalyzer and the spectrophotometric calibration are reprogrammed to meet requirements deriving from urine adoption and to optimize the enzyme assay in this unusual medium.


More on effects of storage time and temperature on urinary enzymes: a 1-year study.

Results of our conclusive study on urinary enzyme stability during sample storage are reported. We measured alanine aminopeptidase (AAP) and N-acetyl-beta-D-glucosaminidase (NAG) in morning urines from 9 healthy normal subjects immediately after collection and throughout a 1-year storage at -70 and -20 degrees C. AAP proved to be quite stable at -70 degrees C (99.2% of the basal value at the end of the year). NAG is partially preserved (84.1% of the basal value) at -70 degrees C, but significantly decreased (50.4%) at -20 degrees C.


Renal thromboxane A2 synthesis in the Lyon hypertensive rat.

The aim of the present study was to assess the mechanisms by which norepinephrine (NE) increased the synthesis of prostanoids and revealed a hyperactivity of the Thromboxane (Tx) A2 synthase in the Lyon genetically hypertensive (LH) rat kidney. To this purpose, the effects of NE (1.2 x 10(-8) to 9.6 x 10(-7) M) on renal function and prostanoid synthesis were assessed in isolated perfused kidneys following beta-adrenoceptor blockade by sotalol (10(-5)M) and compared to those of equipressor concentration of an alpha 2-adrenoceptor agonist, BHT 933 (3.5 x 10(-4) M) and angiotensin II (AII) (7.7 x 10(-9) M). Kidneys were isolated from eight week-old male LH rats and from their normotensive (LN) and low blood pressure (LL) controls and perfused in a single pass system. In baseline conditions, sotalol did not modify renal function or urinary prostanoids in any of the three strains. Following NE stimulation, it potentiated the increase in renal vascular resistance of LL and LN controls but not that of LH rats. The pressure-natriuresis and the urinary prostanoids remained unchanged. BHT 933 elicited a weak stimulation of prostanoid release while AII markedly increased it and revealed, as did NE, the hyperactivity of the TxA2 synthase. It is concluded that the NE-induced stimulation of prostanoid synthesis does not involve beta-adrenoceptors and is unrelated to the associated hemodynamic changes. These results also demonstrate that the increased renal synthesis of TxA2 observed in LH rat kidney is not a specific response to alpha-adrenoceptor stimulation and is likely to involve activation of the phosphoinositide pathway.


Acute effects of hydrocortisone on circulating growth hormone levels in patients with acromegaly.

Aim of our study was to investigate the acute effects of intravenous infusion of hydrocortisone on circulating growth hormone (GH) levels in acromegaly. We studied 5 adult patients with active acromegaly, 3 males and 2 females; age 52 +/- 3.6 years, body mass index 27 +/- 1 kg/m2. The patients underwent in randomized order from 0 to 120 min: (1) intravenous infusion of saline, 250 ml; (2) bolus intravenous injection of hydrocortisone succinate, 100 mg at time 0 followed by intravenous infusion of hydrocortisone succinate, 250 mg in 250 ml of saline for 120 min. Blood samples for GH, cortisol and glucose assay were taken at -15, 0 (time of beginning of saline or hydrocortisone infusion), 15, 30, 45, 60, 90, 120, 150 and 180 min. In all the acromegalic patients, during hydrocortisone succinate infusion, GH values clearly fell with respect to saline (nadir range 18.4-50.5% with respect to baseline levels) with nadir between 60 and 180 min after the beginning of the infusion. Our data show that acute and sustained hypercortisolism, decreases circulating GH levels in acromegaly. It seems likely that also in acromegalic patients as well as in normal subjects short-term increases in serum cortisol levels may be able to cause an enhancement of hypothalamic somatostatin secretion, which in turn may be responsible for the glucocorticoid-mediated GH inhibition.


True hermaphroditism in 45,X/46,XY mosaicism.

This report discusses the clinical findings on two patients with 45,X/46,XY mosaicism, two boys presented with penile hypospadias and cryptorchidism. A dysgenetic ovary and a testis were found in one boy, and a dysgenetic ovary in the other. Both patients can be considered to be true hermaphrodites on the basis of histology and clinical and hormonal observations. 45,X/46,XY mosaics have a wide range of phenotypic appearances and their gonadal morphology can also show great differences. However, the incidence of true hermaphroditism in individuals with 45,X/46,XY mosaicism is low and the reports in the literature rare. It is likely that males with 45,X/46,XY who suffer only mild maldevelopment of the external genitalia will not be recognized. In all patients with penoscrotal hypospadias and cryptorchidism with 45,X/46,XY mosaicism, the possibility of true hermaphroditism should be considered.


The genetics of multiple endocrine neoplasia (MEN).

Multiple Endocrine Neoplasia (MEN) refers to the family of diseases characterized by hyperplasia and/or tumoral proliferation in various organs derived from the neural crest. MEN are transmitted in an autosomal dominant fashion in affected pedigrees with a high degree of penetrance. MEN 1 and MEN 2A/B loci have recently been mapped, respectively, to chromosomes 11 and 10 by linkage analysis using polymorphic DNA markers. These discoveries will lead (1) to a rapid understanding of the physiopathological pathway determining such syndromes and (2) to major clinical impact through the genetic screening.


Pathology of multiple endocrine neoplasias 2A and 2B: a review.

This review describes recent findings on the morphology, function and prognosis of lesions associated with the MEN 2 syndromes. Special emphasis is placed on the analogies and discrepancies between the hereditary and nonhereditary manifestations of the endocrine proliferations involved.


Clinical features of multiple endocrine neoplasia type 1 and type 2.

The term 'Multiple Endocrine Neoplasia' (MEN) denotes a genetically determined syndrome characterized by the independent appearance of benign or malignant changes of several endocrine organs as well as occasional changes of neural, muscular and connective tissue. Three different forms have been identified: MEN 1 (Werner's syndrome) includes parathyroid hyperplasia in combination with pancreatic islet cell and pituitary tumours; MEN 2a (Sipple's syndrome) includes medullary thyroid carcinoma in association with phaeochromocytoma and parathyroid hyperplasia; MEN 2b includes medullary thyroid carcinoma, phaeochromocytoma and mucosal neuromas. MEN syndrome is transmitted as an autosomal dominant trait with a high degree of penetrance. The changes in the individual glands appear to be causally and temporally independent of each other. A spectrum of pathological changes exist in the affected glands which range from hyperplasia to adenoma to carcinoma. The pathological process is almost always multicentric, often resulting in bilateral disease of organs. The appearance of an endocrine tumour known to be associated with MEN should alert the physician to the possibility of a MEN syndrome. When the possibility of such a syndrome exists, screening and long-term observation should be initiated to diagnose a carcinoma in its earliest stage, or before the development of clinical manifestations of hormone excess.


Sensitivity of mosquito-pathogenic bacterial strains to various antibodies.

Four strains of Bacillus sphaericus, 1593, 2362, 9001 and 9002, B. thuringiensis H-14 and B. thuringiensis neoleonensis were tested for sensitivity against 18 antibiotics. The results revealed that all the four strains of B. sphaericus are resistant to colistin, nalidixic acid, polymyxin B and streptomycin. However, B. thuringiensis H-14 was resistant to 9 antibiotics, viz. ampicillin, cephalexin, carbenicillin, co-trimoxazole, colistin, cloxacillin, penicillin, nitrofurantoin and polymyxin B whereas B. thuringiensis neoleonensis was found to be resistant to 8 antibiotics. These results may help in isolation of potential and resistant mosquito pathogenic bacteria.


Genitourinary and sexual adverse effects of psychotropic medication.

We review the adverse effects on genitourinary and sexual function associated with antidepressants, neuroleptics, lithium, and benzodiazepines, and suggest treatment strategies that may be used for their management. This article is based on systematic review of the existing literature, including more than 130 relevant articles on genitourinary and sexual effects of psychotropic medications. We find that genitourinary function, including effects on continence and flow, and sexual function, including libido, erection, ejaculation and orgasm, may be altered by psychotropic administration. Many of these effects may be consequent to the impact of these medications on neurophysiologic systems. Genitourinary and sexual adverse effects associated with psychotropic therapy are important areas of study and clinical concern that may affect patient comfort and compliance with treatment.


The expression of bipolar affective disorders in brain injured patients.

A prospective study was designed to investigate the varied presentations of major affective disorders in patients with organic brain disease. Patients admitted to our neuropsychiatry service, with affective and behavioral disturbances, and known neurological disorders, were classified, on phenomenological grounds, into the following groups: 1) elated mania; 2) irritable mania; 3) affective lability with periods of irritability, but without other symptoms pathognomonic for mania; and 4) intermittent psychosis with absent or ambiguous mood changes. A majority of patients in all four groups responded to pharmacotherapy with anti-cycling agents. It is proposed that these groups represent different expressions of mania in brain injured persons, and that these expressions range through a spectrum of phenomenology, included elated mania, irritable mania, episodic psychosis and explosive organic personality disorder. The DSM-III-R classification of these disorders, and approaches to their clinical management, are discussed.


Interferon-gamma response region in the promoter of the class II MHC gene, DPA.

The class II MHC gene DPA is inducible by interferon-gamma (IFN-gamma), whereas the DQB gene is not inducible in most cell types. To investigate the DNA region specifically responsible for inducibility or its lack that may be required (in addition to the elements required for constitutive expression of class II genes), hybrid promoters were constructed between the proximal 5' regions of the DPA promoter up to -148 bp, which is IFN-gamma inducible, and of the DQB promoter up to -160 bp, which is not inducible. As a result of these and previous studies [9, 10], the region of the DPA gene required for its IFN-gamma inducibility was localized to 27 bp between -55 and -81, including the Y-box element and its flanking nucleotides.


HLA-A and DPB1 loci confer susceptibility to Graves' disease.

To investigate HLA-linked genetic factors involved in the pathogenesis of Graves' disease, 76 patients and 317 healthy controls in the Japanese population were examined for HLA-A, B, C, DR, and DQ specificities by serologic typing and for HLA-DPB1 alleles by DNA typing by using the PCR-SSOP method. The frequencies of HLA-A2, B46, Cw11, and DPB1*0501 were increased and those of HLA-A24, B7, Bw52, and DR1 were decreased in the patients. The increased frequencies of HLA-A2 and DPB1*0501 in the patients were statistically significant when the corrected p value (pc) was applied (pc < 0.02 and pc < 0.002, respectively). ORs for a risk to develop the disease were calculated among individuals positive for DPB1*0501 and/or HLA-A2, and the highest OR (10.5) was observed in individuals possessed both DPB1*0501 and HLA-A2. This observation suggests a synergic involvement of a HLA class II allele (DPB1*0501) and an HLA class I allele (HLA-A2) in the pathogenesis of Graves' disease.


Sequencing and population analysis of four novel HLA-DPA1 alleles.

We determined the base sequences of the HLA-DPA1 gene from four B-lymphoblastoid cell lines (CB6B, LKT3, AMAI, and T7526) that showed distinct electrophoretic patterns of single-stranded polymerase chain reaction products of the HLA-DPA1 gene. The novel HLA-DPA1 alleles of CB6B, LKT3, AMAI, and T7526 were designated DPA1*02021, DPA1*02022, DPA1*0301, and DPA1*0401, respectively. Although there was only one base substitution between DPA1*02021 and DPA1*02022, the single-strand conformation polymorphism of these two alleles was clearly demonstrated by electrophoresis in a nondenaturing polyacrylamide gel containing 10% glycerol. In addition, we genotyped for the HLA-DPA1 gene of healthy unrelated Oriental individuals--i.e., 227 Japanese, 88 Papua New Guineans, and 41 Buyi-Chinese--to demonstrate the ethnic distribution of the HLA-DPA1 alleles.


Characterization of an HLA-DR15 DQ5 haplotype found in the Spanish Caucasoid population.

HLA class II typing by RFLP and PCR-SSOP has been performed on HLA-DR2-positive individuals as a part of a study on MHC in a Spanish Caucasoid population. The results of this study reveal that HLA-DR15 (DRB1*1501 DRB5*0101) and DQ5 (DQA1*0102 DQB1*0501/0502) are not uncommonly associated in such a population. Family segregation has been assessed and allogeneic reactivity against some classic DR2 haplotypes has been tested; a stimulatory capability of DQ6 antigen in this situation is shown. It is suggested that the reported association is not uncommon in European Caucasoids as well as in other populations and it should be considered in matching for transplantation and in DR2-associated diseases.


Laparoscopic cryptorchidectomy in horses.

Laparoscopic cryptorchidectomy was successfully performed in 15 standing or recumbent horses. In 3 horses, owners believed that castrations had been performed, but the horses had retained stallion-like behavior. Successful removal of undescended testes in these horses stopped this behavior. Laparoscopy offered excellent visualization of the structures of the vaginal ring and facilitated removal of the abdominally located testis. The internal and external inguinal rings were not invaded, thus the chance of serious complications that may result during open cryptorchidectomy procedures was minimized.


Dysplastic nevus syndrome: melanoma-prone disease.

Regardless of subsequent clinical courses of patients with dysplastic nevi (DN), substantial evidence supporting DN as one of the melanoma-prone diseases is not yet available, especially in sporadic DN, due to the lack of genetic information other than retrospective studies in clinical observation. This study aimed at the immunohistological characterization of sporadic DN distinct from common nevi (CN) and at the evaluation of the potentially of sporadic DN for malignant transformation. We considered our results together with previous immunological and epidemiological reports. We noted the following three immunohistological characteristics. 1) Proliferating cell nuclear antigen (PCNA), one of the markers for active cell division, could be detected on DN cells in junctional nests of only one among ten DN examined but not on CN cells at all. 2) The altered expression of alpha-smooth muscle actin (alpha-Sma), often observed in melanoma cells, could not be detected in DN cells. However, anti-alpha-Sma monoclonal antibody (MoAb) clearly demonstrated distinctive hypervascularity in the stroma surrounding DN when compared with CN. 3) ME491 antigen, which is known to be expressed mainly in the radial growth phase of melanoma, was detected with similar intensity on both DN and CN. These data indicate that DN has a somewhat higher potentiality than CN for cell division and secretion of some cytokines which can induce hypervascularity in the surrounding stroma, but that DN has not yet undergone the significant phenotypic changes observed in melanoma cells. Further advancements in understanding molecular events in DN cells will be of great benefit in setting DN in the multiple oncogenic spectrum from pigment cells to melanoma.


The interaction between Langerhans cells and CD4+ T cells.

The human skin is increasingly exposed to haptens and environmental protein antigens. Because Langerhans cells represent the outermost network of MHC class II+ antigen presenting cells in mammalians, we investigated their interaction with CD4+ T cells. Hapten-modified Langerhans cells induced proliferation and IL-2 production in naive resting CD4+ T cells. T cells activated in this manner and subsequently cultured with IL-2 mediated contact sensitivity in vivo and produced IL-2 but no IL-4 upon restimulation in vitro. Thus they corresponded to Th1 cells. Repeated stimulation with Langerhans cells induced a modulation of the lymphokine pattern: IL-2- and IL-4-producing Th0-like cells were identified after 3 to 4 rounds of restimulation; after > 5 rounds, Th2-like cells with an IL-4+IL-2- pattern and the capacity for inducing IgE synthesis in B cells was identified. Th2 cells were also recently found to mediate inflammatory tissue lesions containing a cellular infiltrate. This demonstrates that Langerhans cells may activate resting CD4+ T cells, Th1-, Th0- and Th2-like cells. It further shows that Langerhans cells may promote the differentiation of postthymic CD4+ T cells into subsets with distinct immune functions: Th1 cells which have the potential to mediate inflammatory reactions such as allergic contact sensitivity and Th2 cells which may be responsible for abnormalities associated with atopic dermatitis, such as elevated IgE and inflammatory skin lesions containing a cellular infiltrate.


The cornified cell envelope: loricrin and transglutaminases.

The cornified cell envelope (CE) of terminally differentiated human epidermis is a complex structure consisting of several defined protein constituents. The CE is the most insoluble component of the epidermis due to crosslinking by disulfide bonds as well as isodipeptide bonds that are formed by the action of transglutaminases (TGases). We have recently determined that loricrin is the major component of CE. We now have isolated and characterized its gene and showed that it has a simple structure with a single intron. We also show that the loricrin gene maps to position 1q21, which, coincidentally, is similar to the location of the profilaggrin and involucrin genes. Human loricrin in 26 kDa and consists of three long glycine-serine-cysteine rich sequence domains that contain quasi-repeating peptides and which form the novel glycine loop motif. These are interspersed by lysine+glutamine rich domains involved in isodipeptide crosslinks. The glycine loops are thought to be involved in organization of epidermal proteins and maintenance of the flexibility of the epidermis. By use of PCR analyses, we have found that human loricrin consists of two allelic size variants, due to sequence variations in the second glycine loop domain only, and these variants segregate in the human population by normal Mendelian mechanisms. Furthermore, there are multiple sequence variants within these two size class alleles due to various deletions of 12 bp (4 amino acids) in the major loop of this glycine loop domain. In order to study the expression and role of TGases in the formation of CE, we have isolated and sequenced cDNA and genomic clones encoding the TGase1 enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)


Alteration of melanoma melanogenesis by phenotypic modifiers.

Human melanoma cells (MM96E) were incubated with a phenotypic modifier (L-ethionine) to compare its effects on phenotypic expression with those induced by sodium butyrate and dimethyl sulfoxide. In contrast to the latter agents, L-ethionine (8mM) failed to arrest the cell cycle at the G1 phase or to inhibit colony formation ability after 48 hr incubation. Tyrosinase activity changed in parallel with 5-S-cysteinyldopa (5-S-CD) content during treatment with sodium butyrate or dimethyl sulfoxide. Tyrosinase was inhibited in L-ethionine-treated cells, probably because of metabolism of L-ethionine to sulfhydryl compounds; this remains to be clarified. Gamma-glutamyl transpeptidase activity changed inversely with tyrosinase activity after sodium butyrate or dimethyl sulfoxide incubation, whereas L-ethionine did not significantly alter the enzyme activity. In addition, only sodium butyrate induced alkaline phosphatase activity. L-ethionine was less effective than sodium butyrate or dimethyl sulfoxide in inhibiting expression of the B8G3 melanosomal antigen, as determined by Western blotting. These results suggest that phenotypic modifiers (differentiation inducers) affect melanoma cells in various ways and that melanogenesis therefore reflects only one aspect of differentiation in pigment cells.


Comparison of restriction fragment length polymorphisms of ribosomal DNA between Diphyllobothrium nihonkaiense and D. latum.

Restriction fragment length polymorphisms (RFLPs) of ribosomal DNA (rDNA) were compared between Diphyllobothrium latum and D. nihonkaiense using seven kinds of restriction endonucleases. No intra-specific variation in restriction fragment profiles was shown within both species of Diphyllobothrium. Digestion of the genomic DNA with three endonucleases. SmaI, HinfI and HhaI, provided one or two different bands between two species, although the hybridization patterns generated with the others. HindIII, XbaI, StyI and HaeIII, were the same in both. RFLPs in the digested profiles with SmaI, HinfI and HhaI could be used as species-specific markers even if only fragments of strobilae with morphological similarity were available. Other cestodes, Spirometra erinacei and Taenia saginata, used as controls showed quite different restriction fragment patterns with all the enzymes used.


Indoor Air '90: the 5th in a series of international conferences on the indoor environment.

The 5th International Conference on Indoor Air Quality and Climate: INDOOR AIR '90 continued a series of international scientific conferences begun in 1978 on a complex, interdisciplinary subject increasingly recognized to be of importance to human comfort, health and productivity, and having important implications for building design and furnishing, office equipment, appliances, cleaning, heating, ventilating, humidifying and air-conditioning. INDOOR AIR '90 constituted a week long program of 542 paper and poster presentations and forum discussions, 100 exhibits, and a public forum. This paper summarizes some of the highlights of this conference and links these to some of the studies reported at earlier INDOOR AIR Conference.


Bradykinin metabolism pathway in the rat pulmonary circulation.

The contribution made by different enzymes to the degradation of bradykinin in physiological conditions was estimated by examining bradykinin metabolism in rat serum, in the in situ perfused lung and in vivo. Dose-response curves for the hypotensive effect of intra-arterially and intravenously injected bradykinin were obtained in unanaesthetized rats. High-performance liquid chromatography was used to analyse the products of bradykinin breakdown after incubation with rat serum and perfusion through in situ lung preparations. In rat serum, kininase I degraded 34% and kininase II 11% of bradykinin, no evidence for other activities being detected. In the awake rat, D,L-2-mercaptomethyl-3-guanidino-ethylthiopropionic acid, an inhibitor of kininase I, did not reduce the percentage of bradykinin inactivation in the pulmonary circulation. In the in situ perfused lung 65% of bradykinin was metabolized and the main products were BK1-7, BK1-5 and BK4-9. Enalaprilat (an inhibitor of kininase II) blocked the formation of BK1-7 and BK1-5 and increased the recovery of BK4-9. beta-Mercapto-ethanol, which inhibits aminopeptidase P, and diprotin A, a specific inhibitor of dipeptidylaminopeptidase IV, both reduced the formation of BK4-9. Diprotin A also allowed the recovery of BK2-9. Bradykinin degradation and BK4-9 recovery were not affected by endopeptidase inhibitors. Our results show that the main degradation pathway of bradykinin in the lung is through the action of kininase II at the carboxyl terminus, and sequential cleavage by aminopeptidase P followed by dipeptidylaminopeptidase IV at the amino terminus. The amino-terminal degradation of bradykinin represents about 38% of the total lung kininase activity.


Pulmonary kinin metabolism and conversion of angiotensin I in spontaneously hypertensive rats.

To examine the metabolism of kinins and angiotensin I in the pulmonary circulation of spontaneously hypertensive rats (SHR) and normotensive Wistar rats (NWR). Bradykinin inactivation was estimated in vivo by comparison of the hypotensive effect of intra-arterial and intravenous injections, and in the in situ perfused lung by analysing the breakdown products using high-performance liquid chromatography. In vivo pulmonary degradation of bradykinin, but not that of higher homologues of this peptide, was shown to be significantly greater in SHR. Angiotensin I converting activity was found to be increased in lungs of SHR. The recovery of bradykinin and homologues from perfused SHR lung was decreased relative to NWR. Des-(Phe-Arg) fragments of all kinin analogues were identified in the pulmonary perfusates. When bradykinin and des-Arg9-bradykinin were injected in the perfused lungs, the respective fragments 4-9 and 4-8 were also identified in the perfusates. When kininase II was inhibited with enalaprilat, the recovery of bradykinin increased from 10 to 43% in SHR and from 23 to 58% in NWR, whereas about 90% of the higher bradykinin homologues were recovered in both SHR and NWR. Aminopeptidase P and dipeptidylaminopeptidase IV, as measured by the recovery of fragment 4-9 under kininase II inhibition, accounted for about 40% of the total pulmonary kininase activity in the SHR lungs and 25% of that of the NWR lungs. The results show that SHR have increased kininase and angiotensin converting activity compared with NWR, and that kinins as well as angiotensin may contribute to the pathogenesis of hypertension. Aminopeptidase P and dipeptidylaminopeptidase IV may contribute to the increased in vivo degradation of bradykinin observed in the SHR.


[Current alteration of Helicobacter pylori in gastro-duodenal diseases].

The current alteration of the detection rate of Helicobacter pylori (H. pylori) on mucous membranes of the stomach has been surveyed and reviewed for 49 cases of gastro-duodenal diseases. The survey has been conducted for 3 years and an endoscopic examinations have been performed 2-8 times. On the first visit, the diagnosis can be made endoscopically, and H2 blockers and another agents were used as therapeutic drugs. The detection rate was 67.3% (33 cases) for positive cases and positive reaction cases, while the rate was 32.6% (16 cases) in negative cases and negative reaction cases. Among them, the most commonly observed disease was atrophic gastritis, followed by duodenal ulcer, and acute gastric mucous lesions (AGML). Peptic ulcer was not seen. After administration of H2 blockers and anti-ulcer agents, the H. pylori-detection rate was found to fluctuate according to the diseases and alterations in the morbidity period. Some cases indicated the disappearance of H. pylori in accordance with the improvement of the lesion site.


[A study to clarify the mechanism of the usefulness of the macrolides--the influence of clarithromycin to biofilm with P. aeruginosa].

Clarithromycin (CAM) was administered long-term to patients with diffuse panbronchiolitis (DPB) to clarify the mechanism of the usefulness of the macrolides (MLs). 1. A tendency for clinical improvement was observed in 17 patients with DPB. Bacteria were eradicated in 7 of 9 patients with P. aeruginosa found in sputum. 2. A biofilm experimental model with P. aeruginosa was found to be destructed through constant contact with CAM and formed into a single cell with a smooth surface. 3. It was believed that the new lesion forming capability of P. aeruginosa that had been in contact with CAM was reduced due to a significant decrease in adherence to tissue. P. aeruginosa was principally eradicated by the host factors. These results suggested that the improvement in the prognosis of DPB with P. aeruginosa in the sputum after adding MLs was closely related to the destructive effect of the MLs on the biofilm.


Calcium current modulation in frog sympathetic neurones: L-current is relatively insensitive to neurotransmitters.

1. Neurotransmitters (noradrenaline, NA; chicken II luteinizing hormone-releasing hormone, LHRH) and activators of G proteins (GTP-gamma-S and AlF3) partially inhibit calcium current in bullfrog sympathetic neruones. Activation of the remaining current is slowed and shifted to more positive voltages. 2. The N-type calcium current appears to be the type modulated, since approximately 90% of peak current is blocked by omega-conotoxin (omega CgTx) and modulation is not affected by nisoldipine. 3. Calcium current at relatively negative voltages (-30 to -50 mV) is resistant to transmitter modulation. The current at such voltages is also resistant to omega CgTx, suggesting that it results from a different type of calcium channel. 4. The omega CgTx-resistant current includes dihydropyridine (DHP)-sensitive and DHP-resistant components. The omega CgTx- and DHP-resistant current is inhibited by transmitter agonist, but the DHP-sensitive (L-type) current is not. 5. In cells dialysed with a low concentration of calcium buffer (0.1 mM-BAPTA), transmitters still inhibit N-current incompletely. However, L-current was partially inhibited (approximately 10%) by LHRH, NA and the muscarinic agonist oxotremorine-M (OXO-M).


[Recent advances in the etiopathology of chronic B-lymphocytic leukemia].

Considerations to multistep development of B-lymphocytic chronic leukaemia are presented under the following aspects: experimental in vitro cultures of bone marrows B lymphocyte precursors, polyclonal nature of proliferating B lymphocytes in the early stage of CLL, especially in the autoimmune status, the role of viruses, the effect of nonrandom chromosome abnormalities and associated alterations in gene functions, clonal evolution during terminal stages of CLL.


Clinical predictors of 1-year outcome in schizophrenia.

To assess clinical predictors of 1-year outcome in schizophrenia, 63 patients were studied prospectively. Persistent negative and total symptoms after 4 weeks of neuroleptic treatment accounted for 62% of the variance of 1-year outcome, whereas baseline measures showed no relationship to outcome. Thus, 1-year outcome in schizophrenia can be reasonably predicted on the basis of symptoms persisting after 4 weeks of treatment.


Evaluation of the residual efficacy of permethrin-impregnated screens used against mosquitoes in Marigat, Baringo district, Kenya.

Insecticide-impregnated screens and bednets are gradually finding wider use in malaria control programmes. The efficacy of these devices is dependent on the method of application, the acceptability by the people and effectiveness of the insecticide used. The present studies were carried out to determine the duration of the effectiveness of a permethrin-impregnated wall cloth (Mbu cloth) used in the Marigat area of Baringo District, Kenya in order to ensure its effective use. Cotton cloth impregnated with permethrin was hung inside an experimental house in Marigat and small pieces cut off each month for bioassay against mosquitoes over a twelve month period. The wall cloth remained effective for 6, 4 and 10 months against Anopheles gambiae s.l. the known vector of malaria, Culex quinque-fasciatus and Aedes aegypti, respectively.


[Role of N-methyl-D-aspartate receptor in the maintenance of learning-dependent long-term potentiation in rat hippocampal CA3 area].

The effect of microinjection of 2-amino-5-phosphonovaleric acid (APV), a selective NMDA receptor antagonist, into the rat hippocampal CA3 area on the synaptic efficacy and related conditioned behavior during the acquisition and consolidation of discrimination learning behavior was examined. The results showed: (1) After population spike (PS) amplitude had just increased to the maximum through training i.e. learning-dependent LTP had just formed, APV 1 microliter (2 mmol/L) was injected into CA3 area, then the rats were trained during the time of efficacy of the drug in every experimental block. The result demonstrated that the PS amplitude could not be maintained at the highest level but decreased to the pre-experiment level after 8 blocks. Correct response percentage of rats could not be consolidated with further training but decreased to less than 10%. (2) After the PS amplitude had kept up at the highest level, APV 1 microliter (2 mmol/L) was injected into CA3 area, then the rats were trained during the time of efficacy of the drug in every experimental block, in which case the PS amplitude also could not be maintained at the highest level but decreased to the pre-experiment level after 14 blocks. Correlatively, when the correct response percentage of rats decreased gradually to less than 10%, the conditioned response of the animals extinguished, but its extinction speed was slower than it was in result (1). These results suggest that the NMDA receptor in CA3 area plays an important role in the maintenance of the learning-dependent long-term potentiation.


Cloning and disruption of a gene required for growth on acetate but not on ethanol: the acetyl-coenzyme A synthetase gene of Saccharomyces cerevisiae.

A DNA fragment of Saccharomyces cerevisiae with high homology to the acetyl-coenzyme A (acetyl-CoA) synthetase genes of Aspergillus nidulans and Neurospora crassa has been cloned, sequenced and mapped to chromosome I. It contains an open reading frame of 2139 nucleotides, encoding a predicted gene product of 79.2 kDa. In contrast to its ascomycete homologs, there are no introns in the coding sequence. The first ATG codon of the open reading frame is in an unusual context for a translational start site, while the next ATG, 24 codons downstream, is in a more conventional context. Possible implications of two alternative translational start sites for the cellular localization of the enzyme are discussed. A stable mutant of this gene, obtained by the gene disruption technique, had the same low basal activity of acetyl-CoA synthetase as wild-type cells when grown on glucose but completely lacked the strong increase in activity upon entering the stationary phase, providing direct proof that the gene encodes an inducible acetyl-CoA synthetase (ACS1) of yeast. As expected, the mutant was unable to grow on acetate as sole carbon source. Nevertheless, it showed normal induction of isocitrate lyase on acetate media, indicating that activity of acetyl-CoA synthetase is dispensable for induction of the glyoxylate cycle in S. cerevisiae. Surprisingly, disruption of the ACS1 gene did not affect growth on media containing ethanol as the sole carbon source, demonstrating that there are alternative pathways leading to acetyl-CoA under these conditions.


Purification of rat liver arylsulfatase A and its microheterogeneity assayed by crossed affinity-immunoelectrophoresis.

Arylsulfatase A (arylsulfatase sulfohydrolase) EC 3.1.6.1 was purified from rat liver by a procedure consisting of differential centrifugation, Con A-Sepharose and Blue Sepharose chromatography, PBE 94 chromatofocusing, DEAE-cellulose and gel filtration chromatography followed by preparative electrophoresis. A molecular mass of 132,000 was estimated by gradient PAGE. Particular proteins were detected by immunoelectrophoresis. Isoelectric focusing combined with immunoelectrophoresis gave two peaks of arylsulfatase A, with isoelectric points of pH 3.9 and 4.5. Microheterogeneity of rat liver arylsulfatase A was studied by affinity immunoelectrophoresis with 9 different lectins. The presence of concanavalin A-, Lens culinaris agglutinin-, Lotus tetragonolobus agglutinin- and wheat germ agglutinin-reactive forms permitted assessment of the types of carbohydrate moieties in arylsulfatase A.


[Autologous blood-derived stem cell transplantation in the treatment of hematopoietic malignant diseases (Part II)].

Autologous transplantation of circulating stem cells is potentially capable of creating a chance of normal hematopoietic reconstitution in the patients in which both allogeneic and autologous bone marrow transplantation was impossible. The authors have reviewed the possibilities for collection of adequate and sufficient numbers of peripheral blood stem cells as well as up-to-date results of their autografting especially in relation to hematopoietic malignant diseases. The advantages resulting from autologous transplantation of circulating stem cells depends on: 1. The lack of risk of GvH--disease. 2. early and rapid hematopoietic and lymphoid recovery, 3. probably, the low risk of graft contamination by tumor cells in early remission.


Transient eosinophilia in a patient with hairy-cell leukaemia treated with 2-chlorodeoxyadenosine.

Eosinophilia may be associated with various pathologic states such as malignant disorders, atopic and parasitic diseases, certain infections, as well as drug reactions. We describe a case of a 48-year-old patient who was submitted to a single-course therapy with 2-chlorodeoxyadenosine because of hairy-cell leukaemia and previously did not respond to splenectomy. After the treatment marked transient eosinophilia appeared which preceded the achievement of complete remission. It is supposed that eosinophilia might have been caused by the release of cytokines from damaged leukaemic cells.


[Synthesis and pharmacological activities of the 1-substituted 4-methoxycarbonyl fentanyl derivatives].

Five derivatives of 4-methoxycarbonyl fentanyl were synthesized, four of them carry in the molecules some radicals that may alkylate opiate receptors. Their analgesic activities were measured and the irreversible inhibitory effects of all compounds on the electrically elicited contraction of MVD were investigated. The analgesic test showed that the compounds in this series possess analgesic activity with typical morphine--like action. The isolated tissue experiment indicated that these new compounds were all capable of binding with opiate receptors, and that compounds 2, 4 and 5 were determined to be a new irreversible ligand of opiate receptors.


Long-term treatment with carbamazepine affects CSF somatostatin immunoreactivity in epileptic patients.

A number of pharmacological evidence supports the view that somatostatin (SS) may be importantly involved in the seizure susceptibility both in humans and in laboratory animals. In a previous report the Authors have provided the finding that a short-term carbamazepine (CBZ) administration is able to reduce SS-CSF-IR in epileptic patients. The present study has been carried out to investigate whether a long-term treatment with CBZ affects in a similar way SS-IR content in CSF from temporal lobe epileptics (CPS). The results confirm and expand previous evidence suggesting that CBZ lowering effect on CSF-SS-IR may be relevant to its anticonvulsivant action.


Friedreich's disease. A linkage study in southern and central Italy.

We studied linkage and linkage disequilibrium between the genetic locus of Friedreich's disease (FRDA) and two maker loci (D9S15 and D9S5) of chromosome 9q13-q21.1 in 49 subjects from 12 families in southern and central Italy. No recombination event occurred between D9S15 and D9S5, or between these polymorphisms and FRDA. Linkage disequilibrium was not observed between D9S15 or D9S5 or the extended haplotypes and FRDA, but was present between the two polymorphisms.


Genetic linkage analysis and presymptomatic testing in Huntington's disease. First report in Italy.

Since 1979 data about Huntington's Disease (HD) in Campania, a region of Southern Italy, has been collecting. The prevalence of HD in this sample is 30.3 x 10(-6) (115 pedigrees, 1470 individuals). Mean age at onset was 38.67 years and the juvenile (onset before 20 years) accounted for 5.8%. Genetic linkage analysis in 4 unrelated pedigrees with D4S10 and D4S95 DNA probes has been performed. The absence of genetic heterogeneity--already proposed in a cooperative study for one pedigree--has been confirmed in this study.


Proliferative activity of cutaneous melanocytic neoplasms defined by a proliferating cell nuclear antigen labelling index.

To evaluate the proliferative activity of benign, borderline and malignant cutaneous melanocytic neoplasms, 30 cases of malignant melanoma (MM) and 41 cases of naevi were studied by immunostaining using a monoclonal antibody against proliferating cell nuclear antigen (PCNA). PCNA is a nuclear antigen expressed in the late G1 and S phase and serves as a marker of proliferating cells. Invasive MM and MM in situ showed much higher PCNA positivity rates than melanocytic naevi (invasive MM, 18.0%; MM in situ, 11.3%; ordinary melanocytic naevi, 2.6%). The PCNA positivity rate did not increase significantly with the thickness of MM. Among ordinary melanocytic naevi, junctional naevi had a higher PCNA positivity rate than compound or intradermal naevi. Mean PCNA positivity rates for Spitz's naevi and sporadic dysplastic naevi were within the range for ordinary melanocytic naevi, indicating the benign nature of both types of naevus. Contrary to some previous studies, MM in situ showed high proliferative activity, indicating that cells of MM in situ are actively proliferating. This study clearly demonstrates that MM and various types of naevi can be separated according to differences in proliferative activity defined by the PCNA labeling index.


Synthesis and H2-antagonist properties of some 1,2,5-thiadiazole-1-oxide derivatives.

A series of 1,2,5-thiadiazole-1-oxide derivatives has been synthesized and studied for its H2-antagonist properties. These derivatives can be considered derived from classical H2-antagonists in which the structure was deeply modified in order to evidence the minimal structural requirements for the activity. It was found that it is sufficient to have the 1,2,5-thiadiazole-1-oxide ring substituted with an alkylamino moiety and with an aliphatic chain linked to the hydroxy or ether group to achieve compounds as active as cimetidine. A few considerations on the binding on guinea-pig cerebral cortex of a series of H2-antagonists with more and more simplified structures are also reported.


Prodrugs of peptides. 19. Protection of the pyroglutamyl residue against pyroglutamyl aminopeptidase by N-acyloxymethylation and other means.

The N-terminal pyroglutamyl group in several peptides is specifically cleaved by pyroglutamyl aminopeptidase (PAPase I). With the aim of protecting this group against enzymatic cleavage by the prodrug approach, various derivatives of L-pyroglutamyl benzylamide, used as a PAPase I sensitive model pyroglutamyl peptide, were prepared and their stability characteristics determined. The derivatives studied included phenoxycarbonyl, phthalidyl, hydroxymethyl and actoxymethyl derivatives, all formed at the pyroglutamyl NH-moiety. Whereas L-pyroglutamyl benzylamide was rapidly hydrolyzed by PAPase I, all the derivatives were resistant to cleavage by the enzyme. On the other hand, these derivatives, with the exception of the N-phenoxycarbonyl derivative, were readily converted to the parent pyroglutamyl benzylamide by spontaneous or plasma catalyzed hydrolysis, the half-lives of conversion in 80% human plasma being in the range 2.3-8.4 h. The major degradation reaction of the N-phenoxycarbonyl derivative in both buffer and plasma solutions was hydrolytic opening of the pyrrolidone ring. The pH-rate profiles for the degradation of the compounds in aqueous solution were obtained and both specific acid and base catalytic reactions as well as a spontaneous reaction were observed. The results suggest that N-phthalidylation, N-hydroxymethylation and N-acyloxymethylation of pyroglutamyl peptides may be useful prodrug approaches to protect such peptides against cleavage by pyroglutamyl aminopeptidase and hence to improve their delivery characteristics.


Comparison of CD3 and CD2 activation pathways in T cells from young and elderly adults.

The ability of purified T cells to be activated by immobilized anti-CD3 and soluble anti-CD2 monoclonal antibodies (mAbs) was compared using cells from young and old donors. Purified T cells from elderly humans activated with immobilized anti-CD3 mAb incorporated less [3H]thymidine (58,780 vs 92,258 cpm; p < 0.02) into cellular DNA, and secreted less IL-2 into the culture supernatants than did T cells from young donors. In contrast, T cells activated with anti-CD2 mAbs displayed no age-related differences in proliferation or IL-2 production. Anti-CD2 stimulation resulted in equal IL-2 synthesis by cells from young and old donors that was comparable to the amount produced by cells from elderly donors stimulated with immobilized anti-CD3. Northern blot analysis of early cell cycle gene expression by anti-CD2 activated T cells demonstrated no age differences in the expression of p55 IL-2R or c-myc specific mRNA, although T cells from elderly individuals activated with immobilized anti-CD3 showed statistically significant decreases in both mRNAs. T cell receptor beta chain mRNA levels did not differ between cells from young or old donors after activation by either anti-CD3 or anti-CD2. The discordance in proliferative ability, IL-2 secretion, and specific mRNA expression between T cells from elderly donors activated through the CD3-TCR complex or by soluble anti-CD2 mAbs provides additional evidence for a multifactorial causation of age-related T cell proliferative defects, and may indicate that the difference in proliferative ability is, in part, attributable to responsiveness to secreted IL-2.


[Functionally important residues of glutamic acid in E. coli pyrophosphatase. I. Chemical modification and localization in the primary structure].

Inorganic pyrophosphatase of E. coli is rapidly and irreversibly inactivated by 5-ethyl-5-phenylisoxazolium-3'-sulfonate (Woodward's reagent K). The appearance in the absorption spectrum of a maximum at 340 nm testifies to the formation of an enzyme enol ester with the inhibitor. The non-hydrolyzable substrate analog CaPP1 partly protects the enzyme from inactivation. A peptide has been isolated from a tryptic hydrolysate of inactivated enzyme which contains an amino acid residue whose modification is critical for the enzyme activity. This peptide corresponds to residues 95-104 of pyrophosphatase and contains four dicarboxylic acid residues. A peptide containing a modified glutamic acid residue was isolated from modified pyrophosphatase hydrolyzed by protease v8. This peptide represents a fragment of a tryptic modified peptide and has a Glu-Ala-Gly-Glu (residues 98-1C1) structure. It is concluded that inactivation of E. coli pyrophosphatase by Woodward's reagent K is a result of selective modification of Glu98, apparently by the most reactive dicarboxylic amino acid within the enzyme active center.


Thunberginols C, D, and E, new antiallergic and antimicrobial dihydroisocoumarins, and thunberginol G 3'-O-glucoside and (-)-hydrangenol 4'-O-glucoside, new dihydroisocoumarin glycosides, from Hydrangeae Dulcis Folium.

New antiallergic and antimicrobial dihydroisocoumarins, thunberginols C, D, and E, were isolated from Hydrangeae Dulcis Folium, the fermented and dried leaves of Hydrangea macrophylla SERINGE var. thunbergii MAKINO, together with new dihydroisocoumarin glycosides, thunberginol G 3'-O-glucoside and (-)-hydrangenol 4'-O-glucoside. Their chemical structures have been determined on the basis of chemical and physicochemical evidence. Thunberginols C, D, E, G, and (-)-hydrangenol 4'-O-glucoside showed antiallergic activity in the in vitro bioassay using the Schults-Dale reaction in sensitized guinea pig bronchial muscle, and they also exhibited antimicrobial activity against oral bacteria.


Anti-eye muscle antibodies and hypothyroid Graves' disease: a case report.

We report the case of a 70-year-old man who developed hypothyroidism associated with TSH receptor antibodies and severe ophthalmopathy during lithium therapy. He had received lithium therapy for more than 20 years for manic depression, when ophthalmopathy (class VI of the American Thyroid Association classification) and mild hypothyroidism developed. Orbital magnetic resonance imaging indicated marked enlargement of the superior, medial and inferior rectus muscles in the left eye. He had anti-eye muscle antibodies in his serum, detected by Western blotting and quantified by chromatoscanning, as well as anti-TSH receptor antibodies. He was treated with supplementation of levothyroxine and four cycles of methylprednisolone pulse therapy. After the pulse therapy, both anti-eye muscle antibodies and anti-TSH receptor antibodies decreased and disappeared in parallel with the improvement in eye symptoms and signs. These observations suggest the importance of anti-eye muscle antibodies as clinical markers in the development of thyroid-associated ophthalmopathy.


Hemorrhagic fever with renal syndrome in Yugoslavia: epidemiologic and epizootiologic features of a nationwide outbreak in 1989.

A nationwide epidemic of hemorrhagic fever with renal syndrome (HFRS) occurred in Yugoslavia in 1989. Sera from 609 hospitalized patients, from all six Republics (Bosnia and Hercegovina, Croatia, Macedonia, Montenegro Serbia, Slovenia) and two Provinces (Kosovo and Vojvodina), who had signs and symptoms suggestive of HFRS, and sera and lung tissues from 544 small mammals belonging to 13 species were studied for evidence of hantavirus infection. Of the 226 patients with serologically confirmed HFRS, 182 resided in Bosnia and Hercegovina or in Serbia. The severity of disease differed from region to region, with an overall fatality of 6.6% (15/226). Patients from southern Yugoslavia tended to have more severe disease and exhibited two types of antibody patterns, while approximately equal numbers of clinically severe and mild cases of HFRS were registered in central Yugoslavia, where four types of antibody patterns were found. Two of these antibody patterns suggested the existence of hantaviruses which are antigenically distinct from those reported to date. Two seasonal peaks of disease, one during the summer and the other in late autumn, were found. Hantaviral antibodies and/or antigens were detected most often in the yellow-necked mouse (Apodemus flavicollis) (88/189), the wood mouse (Apodemus sylvaticus) (28/146), the striped field mouse (Apodemus agrarius) (10/64), the bank vole (Clethrionomys glareolus) (36/63), the house mouse (Mus musculus) (14/29), and the Norway rat (Rattus norvegicus) (14/21). Five other species of rodents and insectivores were infrequently infected.


[A case of prenatal diagnosis of beta-thalassemia by polymerase chain reaction].

The prenatal diagnosis of beta-thalassemia in the Udin family, where the parents were the carriers of 2 bp deletion in the codon 8 (-AA) was undertaken using PCR. Five polymorphic restriction endonuclease sites in the beta-globin gene region were tested. They are: 2 HindIII sites in the gamma G and gamma A genes, 2 HincII sites located in the pseudogene and in its 3'-flanking region, and the AvaIII site in the second exon of the beta-globin gene. The heteroduplex analysis was also performed. Two HindIII polymorphic sites were informative and the HincII site in the pseudogene and the AvaII site in the beta-globin gene were partially informative. According to the results of the RFLP analysis, the embryo was heterozygous. The similar result was obtained by heteroduplex analysis.


Plasma fibrinogen levels and fibrinogen genotype in non-insulin dependent diabetics.

The association between plasma fibrinogen levels, fibrinogen genotype, and the development of macrovascular disease was studied in 100 patients with non-insulin dependent diabetes mellitus (NIDDM). The mean plasma fibrinogen levels in patients with macrovascular disease was higher than those without, although the difference was not statistically significant (3.67 g l-1, and 3.43 g l-1, respectively). The frequency of the rare allele of the fibrinogen gene DNA polymorphism detected with the restriction enzyme Bc1I was slightly higher in the group of patients with disease, but the difference was not statistically significant (0.20 vs 0.16). The frequency of the TaqI polymorphism rare allele was the same in both groups (0.30 vs 0.31). However, the Bc1I polymorphism was strongly associated with plasma fibrinogen levels, with those patients heterozygous for the rare allele having mean levels 16 per cent higher than those lacking the allele (3.81 g l-1 vs 3.28 g l-1, p < 0.05). This data demonstrates that variation at the fibrinogen locus is involved in determining fibrinogen levels in patients with NIDDM, and suggests the possibility that fibrinogen genotype and plasma fibrinogen levels could be one of the factors making a small contribution to the development of macrovascular disease in diabetic patients.


Activation of CD4+ and CD8+ lymphocyte subsets by streptozotocin in the popliteal lymph node assay. II. Comparison with acute graft-vs-host reaction in H-2 incompatible F1 mouse hybrids.

Changes in lymphocyte subsets during an acute GVH reaction were compared to STZ-induced PLN response in mice. The GVH reaction was induced locally by sc injection of parental C57Bl/6 [B6] spleen cells into (C57Bl/6 x DBA/2) F1 footpad [B6D2F1]. Early cell activation and time-related changes in T- and B-lymphocyte subsets were monitored during the onset of the GVH reaction by flow cytometry and immunophenotyping. Examination of cell size and chromatin decondensing for T- and B-cell subsets showed differences in activation profile during the early phase of the GVH reaction. The present study provides direct evidence for early in vivo activation of both CD4+ and CD8+ T-cells. Our data confirm the central role of T-cell activation in the induction of a GVH reaction and suggest that recirculatory host B-cells can play an important role in early GVH node enlargement. Overall, our comparative analysis supports the concept of polyclonal T-cell activation for both STZ-related and GVH-induced lymphoproliferation. Chemicals-induced lymphoproliferation leading to autoimmune reactions is a challenging issue. A number of drugs and chemicals have been tested in the PLNA assay for lymphoproliferative potential. We previously reported the activation and proliferation of T-cell subsets following STZ injection into murine footpads. The STZ-induced PLN enlargement and proliferation characteristics of T- and B-cell subsets were postulated to be similar to those of an acute allogeneic GVHR. In the present study, a cytometric analysis of T- and B-cell subsets in PLNs was performed during an acute allogeneic GVHR, for comparison purposes. Such a reaction results in a massive node enlargement five to ten times that seen after stimulation with conventional antigens. Acute GVHR is believed to be a direct consequence of the high frequency of alloreactive donor T-cells inducing a massive proliferation of B-cells, almost exclusively of host origin, in GVHR nodes. It is now widely accepted that donor T-cells activated as the result of exposure to foreign MHC antigens in the recipient, secrete various cytokines which assist the host B-cells and bypass the normal B-T cell cooperation. Induction of an acute GVHR, as in the parental B6--->recipient B6D2F1 model, requires the injection of CD4+ and CD8+ donor T-cells into an F1 recipient that differs from the parent at both MHC class I and II loci.(ABSTRACT TRUNCATED AT 400 WORDS)


Ethanol-induced CD3 and CD2 hyporesponsiveness of peripheral blood T lymphocytes.

The functional relevance of a direct ethanol effect on the membrane structure of T lymphocytes and accessory cells (APC), as well as on signal transduction systems was studied in ten normal subjects. Ethanol incubation (80 mM for 24h) of highly purified T cells increased the number of CD4+/CD45RA+ lymphocytes. In contrast, ethanol exposure induced a drop in CD14+/LFA-3+ APC values. These changes were accompanied by faulty T-cell proliferation in response to anti-CD3 and anti-CD2 mAb and inhibition of CD3- and CD2-mediated rises in intracellular calcium and, to a lesser extent, inositol 1,4,5-triphosphate levels. These data clearly indicate that a membrane-specific ethanol interaction both modifies surface glycoproteic and/or glycolipidic structures and alters transmembrane transduction of the activation signals.


Persistent depletion of CD4+ T cells and inversion of the CD4/CD8 T cell ratio induced by anti-CD4 therapy.

A 49-year-old patient with refractory rheumatoid arthritis was treated repeatedly with anti-CD4 murine monoclonal antibodies. While the first anti-CD4 treatment resulted in a marked, however transient, depletion of CD4+ cells from 1070 to a minimum of 175/microliters, a second treatment cycle resulted in a persistent decrease. Despite this marked depletion, no major clinical improvement occurred, which was in striking contrast to other patients treated in a similar way. Of interest, the administration of low doses of chlorambucil led to significant clinical benefits. Markedly reduced numbers of CD4+ cells (200-500/microliters) were observed for more than 2 years, while the numbers of CD8+ cells increased after the second treatment. No infectious episodes occurred. Discontinuation of chlorambucil did not lead to increasing amounts of CD4+ cells. In contrast to the rapid reduction of CD4+ cells from the blood stream induced by anti-CD4 infusions, there was a considerable delay until altered CD4/CD8 ratios were observed in intraarticular sites. No evidence was found for either humoral or cellular immune reactivities towards CD4+ T helper cells. Our findings suggest that in certain patients undergoing anti-CD4 therapy there may be a reduced capacity of the CD4+ T helper cell pool to regenerate.


[Peripheral blood IgA bearing cells in children with Henoch-Schönlein purpura nephritis and IgA nephropathy. Relationship between IgA bearing cells and clinico-pathological findings or T alpha 4 cells].

It has been reported that patients with Henoch-Schönlein purpura nephritis (HSPN) and IgA nephropathy (IgAN) showed an familial increase of IgA bearing peripheral blood lymphocyte. To elucidate the relationship between IgA bearing cells and clinico-pathological findings or T cell subsets, especially IgA specific helper T cells (T alpha 4 cells), 20 patients with HSPN and 33 patients with IgAN were studied. The results demonstrated as follows; 1) IgA bearing cells were significantly increased in patients with both HSPN and IgAN (p < 0.001). 2) The increase of IgA bearing cells was well correlated to the degree of patients' proteinuria and hematuria (p < 0.05), and also correlated with the severity of patients' renal pathological findings in both diseases. 3) With relation to the T cell subsets in patients with both diseases, only the CD4+ Leu8- cells (helper T cells) and T alpha 4 cells were significantly increased, in addition, positive correlation between IgA bearing cells and CD4+ Leu8- cells or T alpha 4 cells was observed. 4) The increase of IgA bearing cells seemed to be transient in HSPN, but remained elevated in IgAN. In conclusion, it was indicated that patients with HSPN and IgAN could have IgA related immunological abnormalities, which may be reflected in the increase of IgA bearing cells and T alpha 4 cells. It was also suggested that determination of IgA bearing cells could be a useful parameter which may be reflected in the disease activity of HSPN and IgAN.


A simple method for electrical field stimulation of cultured granule cells.

A method suitable for electrical field stimulation of superfused primary cultures of cerebellar granule cells is described. A microchamber of about 0.5 ml was obtained by closing the culture dishes with a perspex plug equipped with stimulating electrodes and inlet-outlet tubing. Two-minute trains of electrical pulses (alternate polarity, 2-ms duration; 100 mA intensity; 10 V drop between electrodes; frequency 5, 10 and 20 Hz) applied to cultures kept at 27 degrees C, elicited a D-[3H]aspartate outflow which was frequency related, [Ca2+]0 dependent, tetrodotoxin sensitive. Moreover 2 trains of 10 Hz pulses (S1 and S2) at 30-min intervals caused an S2/S1 ratio equal or near to one, thus demonstrating that a steady-state condition had been achieved. The NMDA antagonist DL-2-amino-5-phosphonopentanoic acid (AP5) but not the non-NMDA antagonist, 6-cyano-7-nitroquinozaline-2,3-dione (CNQX), added before S2, significantly increased the electrically evoked tritium efflux, suggesting that the endogenous transmitter released during electrical stimulation activated an NMDA-mediated negative feed-back. This technique of electrical field stimulation seems particularly feasible to study the extent and time course of drug effects on spontaneous and evoked D-[3H]aspartate outflow.


Fast in vivo monitoring of electrically evoked dopamine release by differential pulse amperometry with untreated carbon fibre electrodes.

Differential pulse amperometry has previously been used in combination with electrochemically treated carbon fibre electrodes. In order to improve the time resolution, this technique was combined here with untreated electrodes. Dopamine release was evoked in the nucleus accumbens of rats anaesthetized with urethane by electrical stimulation of the mesolimbic dopaminergic pathway. The differential oxidation current appearing at +200 mV was recorded every 1 s and was proportional to the dopamine concentration from 0.5 to 50 microM. At this voltage these untreated electrodes were not sensitive to the main catechol metabolite (DOPAC) and poorly sensitive to ascorbic acid. The electrically evoked increase in the oxidation current corresponded exclusively to dopamine. It was enhanced by nomifensine, amphetamine, haloperidol and pargyline and reduced by alpha-methyl-p-tyrosine (A-MPT). The results show that the evoked DA release was facilitated by increasing the stimulation frequency from 10 to 40 Hz. The method was sufficiently sensitive to detect dopamine release evoked by electrical stimulation at 10 Hz and its time resolution was 1 s.


[Telethermography in the examination of children following orchiopexy].

From the results of examination of 60 healthy boys the authors suggest a thermographic characteristics of the scrotum in different age groups. It is shown with high authenticity that the temperature of the scrotum reduces with age. Besides, there is an authentic increase of temperature of the scrotum on the left side in children aged from 11 to 14 years. Examination of 317 boys who underwent operation for cryptorchidism showed the temperature to be increased by 0.6-0.8 degree C in the early and by 0.1-0.4 degree C in the late-term postoperative period. Distant thermography was found to be highly informative in studying the scrotal temperature regimen in healthy children and in those with cryptorchidism in the postoperative period.


Determination of release of endogenous dopamine from superfused substantia nigra slices.

Dopamine (DA) is synthesized and released not only from the terminals of the nigrostriatal dopaminergic neuronal pathway but also from cell bodies and dendrites in the substantia nigra (SN). In most of the in vitro studies on DA release in the SN, release of exogenously applied 3H-DA has been determined either from slices or from synaptosomes. However 3H-DA has some disadvantages; 3H-DA is taken up and released not only from dopaminergic cells but also from other neuronal elements and radiolabelled DA may not be evenly distributed with the releasable endogenous pool of the amine. Therefore we have developed a method for determination of the release of endogenous DA from superfused guinea pig SN slices. We have used a superfusion system in which 6 slices are placed into a microchamber. Samples of superfusate were collected every 10 min for up to 3 h and biochemical analyses were performed by electrochemical detection preceded by absorption of DA on alumina and chromatography on a cation exchange column. Expected increases of the release of endogenous DA were obtained following D-amphetamine and potassium administration. The data indicate that it is possible to measure endogenous release of DA from guinea pig SN slices with standard HPLC technique and follow the release for a relatively long time.


A simple biological way to screen dopaminergic agonists.

The velocity of propagation of the in vitro retinal model of spreading depression is very sensitive to changes in the ionic composition of the extracellular medium and also to the the addition of different drugs. 10 microM of SKF 38393, a D1 agonist, increases the velocity of propagation of the wave while 10 microM of Quinpirole, a D2 agonist, decreases it. Both changes are blocked by their specific antagonists, SCH23390 and 1-sulpiride respectively. This assay can biologically screen potential dopaminergic drugs indicating its physiological D1 and/or D2 preferred effect in the tissue for future analysis by other different methodologies.


Somatostatin and its analogs in the short bowel syndrome.

The use of somatostatin to manage diarrhea associated with the short gut syndrome is impractical because of its need to be given by continuous infusion and a rebound effect on stool output with cessation of therapy. Octreotide has been used more successfully to control stool and electrolyte losses in patients with shortened gastrointestinal tracts. In published series and studies, all subjects appear to decrease stool losses, but clinical benefit for long-term use is not achieved for all patients. In the patients who do respond, the need for parenteral nutrition and intravenous hydration has been decreased or eliminated. The optimal dose is unclear, but many patients respond to 50-micrograms injections twice daily. Several investigations noted no additional beneficial effects with escalating dosages. Adverse effects include impairment of fat absorption, which may offset the therapeutic benefits of octreotide. The patients with the greatest response appear to have the least fat malabsorption. Other adverse effects noted when using octreotide for control of the short gut syndrome include pain associated with subcutaneous injection and abdominal complaints. Other potential concerns include the effect on gallstone formation in this high-risk population and intestinal adaptation.


[A comparative analysis of the antihypertensive activity of calcium antagonists and adrenoblockers in long-term treatment].

Essential hypertension stage II was treated by calcium antagonists, alpha- and beta-adrenoblockers in 362 patients. The drugs were compared for hypotensive efficiency at rest and exercise. A significant hypotensive effect was achieved in 81%, 58%, 44%, 43% and 37% of patients treated with labetalol, nifedipine, nadolol, propranolol, diltiazem, respectively. An increase in verapamil daily dose from 240 to 600 mg led to a rise in efficacy from 27 to 75% without an increase in the number of side effects. The antihypertensive effect of the drugs is shown to persist in long-term treatment as shown by exercise tests and static loads.


[Cardiovascular effects of a combination of vecuronium and low-dose fentanyl in atropinized and non-atropinized subjects].

The cardiovascular effects of the pharmacologic association of low-dose fentanyl (2 micrograms/kg) and vecuronium (120 micrograms/kg) have been studied in 38 ASA I and II atropinized and non-atropinized patients scheduled for abdominal surgery during induction of anaesthesia with thiopentone or propofol. Whatever the induction agent used, heart rate was consistently reduced in patients not receiving an anticholinergic drug, while it was unchanged in patients treated with atropine intravenously. In non-atropinized patients impressively lower minimum heart rates were observed during induction of anaesthesia with thiopentone. In this last group one patient suffered from a cardiac arrest resolved without sequelae. In patients treated with the association between vecuronium and low doses of fentanyl a pretreatment with atropine is always indicated. Propofol seems to be a better induction agent than thiopentone.


Differential in vivo and in vitro effect of gentamicin on glutamate synthesis and glutamate deamination in rabbit kidney-cortex tubules and mitochondria.

The effect of gentamicin on both glutamate synthesis and glutamate deamination was studied in kidney-cortex mitochondria and tubules isolated from both control and gentamicin-treated animals. In kidney-cortex mitochondria which were permeabilized in order to make a free access of substrates and antibiotic to the glutamate dehydrogenase, gentamicin appeared to be a very potent inhibitor of glutamate synthesis, resulting in about 60% decrease of the enzyme activity at 5 mM concentration. Other aminoglycoside antibiotics decreased the enzymatic activity, in the following order: gentamicin > neomycin = tobramycin = kanamycin > biodacyna > amikacin > streptomycin. This, in principle, corresponds to their known nephrotoxic potential observed in vivo. The inhibitory action of antibiotics was abolished by neither ADP nor leucine, allosteric activators of glutamate dehydrogenase. Surprisingly, gentamicin did not decrease the rate of ammonia formation from glutamate when added to both renal tubules and mitochondria isolated from control rabbits. This indicates that the antibiotic exerts its inhibitory effect on glutamate dehydrogenase activity in the direction of glutamate synthesis only. In contrast, the rate of both glutamate deamination and glutamate synthesis was about 40% lower in renal tubules and mitochondria isolated from kidney-cortex of animals which were given antibiotics for 10 days. In view of these results it seems that (i) the depression of ammoniagenesis in gentamicin-treated animals may be due to a decrease of glutamate dehydrogenase content and (ii) under conditions in vitro the aminoglycoside inhibits the enzyme activity in the direction of glutamate synthesis while it does not affect the glutamate deamination.


K-opioid receptor changes in experimental models of cerebral ischaemia and atherosclerosis in the rabbit.

Thromboembolic phenomena and transient ischaemic attacks (TIA) are considered the basis of ischaemic pathologies. The aim of the present research is to investigate the involvement of k-opioid receptors in cerebral blood flow (CBF) impairment which results in experimental stroke or dietary atherosclerosis in rabbits. CBF measurement showed a significant decrease in rabbits submitted to embolization and/or atherosclerosis. Binding studies showed that massive cerebral ischaemia and atherosclerosis produced a significant increase in the number of k-opioid receptors (Bmax), without changing (KD) affinity values. In conclusion, the results obtained seem to indicate that the increase in k-opioid receptors might play a crucial role in a common cerebral biochemical mechanism both in ischaemic and atherosclerotic pathologies.


Unexplained hypertension during induction of a patient with phaeochromocytoma.

An unexplained hypertensive response during the induction of anaesthesia for phaeochromocytoma resection is described. This response was not accompanied by elevations in plasma catecholamine levels. It occurred despite heavy premedication and followed induction with etomidate, alfentanil, lignocaine, vecuronium and magnesium sulphate (MgSO4). A bolus of esmolol (0.5 mg/kg) lowered the blood pressure rapidly. Subsequent haemodynamic manipulations were carried out by varying the rate of an esmolol infusion. A constant background infusion of MgSO4 was maintained throughout the procedure. These produced satisfactory haemodynamic control despite marked rises in plasma catecholamine levels.


Methotrexate increases valproic acid-induced developmental toxicity, in particular neural tube defects in mice.

The hypothesis that valproic acid-induced dysmorphogenesis may be due to an interference of this drug with folate metabolic pathways was further investigated by a study of a possible interaction of valproic acid (VPA) and the established folate antagonist methotrexate (MTX). The dihydrofolate reductase inhibitor MTX (1.25 and 2.5 mg/kg, i.p.) was injected 15 min prior to VPA (300 and 400 mg/kg, s.c.) in day 8 pregnant NMRI mice. Fetuses were examined for exencephaly, resorption, and fetal weight retardation on day 18 of gestation. MTX produced no exencephaly or reduction in fetal weight, and the 2.5-mg/kg dose caused 56% resorption. Higher doses (5-20 mg/kg) produced embryolethality and fetal weight retardation, but no exencephaly. VPA (300 and 400 mg/kg) administration resulted in 3.4% and 12.6% exencephaly and 9% and 19% resorptions, respectively. Coadministration of MTX with VPA significantly increased VPA-induced resorption and exencephaly rates as well as fetal weight retardation. Exencephaly induced by VPA 400 mg/kg was increased to 29.5% and 24.1% (P < 0.01 and P < 0.05) when given with 1.25 and 2.5 mg/kg MTX, respectively. MTX (2.5 mg/kg i.p.) did not alter transplacental VPA (400 mg/kg, s.c.) pharmacokinetics. These results support the view that VPA-induced teratogenesis may be mediated by interaction with folate metabolism.


Teratological study of stobadin after single and repeated administration in rats.

The toxic developmental potential of the anti-arrhythmic drug stobadin was assessed after single intravenous or repeated oral doses to pregnant rats. Stobadin was studied in the form of dihydrochloride (DH 1011) at doses of 2 and 6 mg/kg, given in single intravenous injections on days 3, 6, 9, or 12 of gestation. Immediately after injection of the 6-mg/kg dose of DH 1011 to pregnant rats, saccade abdominal respiration, tremor of hindlimbs, and sedative behaviour were observed on each day of medication. No deaths of females occurred in either the control or experimental groups. Slight foetal toxicity was manifested by significantly decreased foetal weight only after treatment on day 3 of gestation at 6 mg/kg and by significantly increased incidence of delayed ossification of the parietal and supraoccipital bone also at 6 mg/kg DH 1011 given on day 12 of gestation. The effect of repeated oral treatment in the form of dipalmitate salt (DP 1031) was studied in doses of 5, 15, and 45 mg/kg from days 2-15 of gestation. Oral exposure to 45 mg/kg DP 1031 resulted in significant reduction of maternal body weight gain and in embryofoetal toxicity, namely, increased preimplantation foetal loss, anomalies of sternebrae, and, after 15 and 45 mg/kg DP 1031, significantly decreased foetal weight and smaller litter size. The relevance of the two routes of stobadin administration for risk extrapolation is discussed.


Transplacental genotoxicity of triethylenemelamine, benzene, and vinblastine in mice.

Transplacental cytogenetic effects of triethylenemelamine (TEM), benzene, and vinblastine on maternal mice and their fetuses have been investigated using micronucleus and sister chromatid exchange (SCE) as genetic endpoints. CD-1 mice were treated on day 14 and 15 of gestation with TEM (0.125, 0.25, and 0.5 mg/kg), benzene (439,878, and 1,318 mg/kg), and vinblastine (0.5, 1, and 2 mg/kg) by intraperitoneal injection at 24 hr intervals, and sacrificed 40 hr after the first injection. Erythrocytic precursor cells in maternal bone marrow and fetal livers (2-4) from each pregnant mouse were used for the micronucleus and/or the SCE analyses. Significant dose-related increases in both micronuclei and SCE were found in maternal bone marrow and fetal liver following TEM treatment. Benzene at the highest dose (1,318 mg/kg) also caused a significant increase in micronuclei and SCE in both maternal bone marrow and fetal liver cells. The embryonic genotoxic effect of TEM was much higher than that of benzene for both genetic endpoints, and the frequency of micronuclei induced by benzene was higher in fetal liver than in maternal bone marrow cells. Vinblastine, a spindle poison, induced micronuclei but not SCE. Micronuclei induction by vinblastine was 7 fold greater in maternal bone marrow than in fetal liver cells. All three chemicals were cytotoxic in maternal bone marrow cells, but not in fetal liver cells except for TEM, which showed a weak cytotoxicity in fetal liver cells in the micronucleus assay. These results indicate that TEM, benzene, and vinblastine are transplacental genotoxicants in mice.


Changes in secondary structure of DNA of rat embryos following treatment with diethylnitrosamine and methylazoxymethanol acetate in vivo.

Diethylnitrosamine (DEN) and methylazoxymethanol acetate (MAM) are not transplacental carcinogenic but embryotoxic to Wistar rats when administered by i.p. injection on day 12 of gestation. MAM, a weak teratogen to rats during this period, induced a dose dependent increase in the number of resorptions to 15% and 40% of the litters following doses of 15 and 25 mg/kg bw, respectively. Rats similarly treated with 70, 150, and 180 mg DEN/kg bw resulted in increases in total DNA mass of day 13 embryos by 31%, 45% and 52%, respectively, compared to the saline treated controls. Twenty percent reduction in total DNA amount was detected following 25 mg MAM/kg bw. Benzoylated DEAE-cellulose (BD-cellulose) chromatography fractionates DNA on the basis of secondary structure by stepwise elution of double-stranded DNA with 1.0M NaCl solution (SE-DNA) followed by elution of DNA containing single-stranded regions with caffeine solution (CE-DNA). Day 13 embryonic DNA was monitored by in vivo labelling with [methyl-3H]-thymidine (3H-TdR) on days 6 and 7 of gestation. Significant increases in percentages of caffeine-eluted DNA (%CE-DNA) compared to control values were detected 24 h after treatment of day 12 embryos with 70, 150, and 180 mg DEN/kg bw. Such increases were not observed after MAM. Incorporation of [methyl-14C]-thymidine (14C-TdR) into embryonic DNA demonstrated the effects of treatment with these compounds on DNA synthesis in vivo. When compared to saline controls, DEN induced significant increases in 14C-TdR incorporation into embryo DNA, 1 h prior to analysis, but the increases were not proportional to the doses administered. Similar analysis of MAM treated samples showed no significant changes to %CE-DNA values. The relative %CE-DNA is expressed as the ratio of the percentage of caffeine-eluted 14C-labelled DNA to %CE-DNA (i.e., %CE-14C-DNA:%CE-3H-DNA). In the majority of control embryos the 14C-specific activity of CE-DNA was higher than the 14C-specific activity of SE-DNA. No significant change to relative %CE-DNA values of embryos to those of the controls was observed 24 h after treatment of day 12 gestation rats with single doses of DEN and MAM. The results of this study support the hypothesis that initiation mechanisms of teratogenesis and transplacental carcinogenesis are different. The pertinence of %CE-DNA and relative %CE-DNA values to teratogenesis and transplacental carcinogenesis is also discussed.


A comparison of an enzymatic and a gas-chromatographic method for measuring the acetate concentration in the blood plasma of cattle.

The concentration of acetate in blood plasma was measured by both a gas-liquid chromatographic method and an enzymatic method using acetyl-CoA synthetase. When acetate was measured enzymatically without previous protein precipitation, the apparent concentration was lower than when the concentration measured by either a gas-chromatographic method or by the enzymatic method after protein precipitation with perchloric acid and neutralization.