Transforming growth factor-beta activity is potentiated by heparin via dissociation of the transforming growth factor-beta/alpha 2-macroglobulin inactive complex.

The control of smooth muscle cell (SMC) proliferation is determined by the combined actions of mitogens, such as platelet-derived growth factor, and the opposing action of growth inhibitory agents, such as heparin and transforming growth factor-beta (TGF-beta). The present studies identify an interaction between heparin and TGF-beta in which heparin potentiates the biological action of TGF-beta. Using a neutralizing antibody to TGF-beta, we observed that the short term antiproliferative effect of heparin depended upon the presence of biologically active TGF-beta. This effect was observed in rat and bovine aortic SMC and in CCL64 cells, but not in human saphenous vein SMC. Binding studies demonstrated that the addition of heparin (100 micrograms/ml) to medium containing 10% plasma-derived serum resulted in a 45% increase in the specific binding of 125I-TGF-beta to cells. Likewise, heparin induced a twofold increase in the growth inhibitory action of TGF-beta at concentrations of TGF-beta near its apparent dissociation constant. Using 125I-labeled TGF-beta, we demonstrated that TGF-beta complexes with the plasma component alpha 2-macroglobulin, but not with fibronectin. Heparin increases the electrophoretic mobility of TGF-beta apparently by freeing TGF-beta from its complex with alpha 2-macroglobulin. Dextran sulfate, another highly charged antiproliferative molecule, but not chondroitin sulfate or dermatan sulfate, similarly modified TGF-beta's mobility. Relatively high, antiproliferative concentrations of heparin (1-100 micrograms/ml) were required to dissociate the TGF-beta/alpha 2-macroglobulin complex. Thus, it appears that the antiproliferative effect of heparin may be partially attributed to its ability to potentiate the biological activity of TGF-beta by dissociating it from alpha 2-macroglobulin, which normally renders it inactive. We suggest that heparin-like agents may be important regulators of TGF-beta's biological activity.

The pharmacological chaperone 1-deoxygalactonojirimycin increases alpha-galactosidase A levels in Fabry patient cell lines.

Fabry disease is an X-linked lysosomal storage disorder caused by mutations in the gene encoding alpha-galactosidase A (alpha-Gal A), with consequent accumulation of its major glycosphingolipid substrate, globotriaosylceramide (GL-3). Over 500 Fabry mutations have been reported; approximately 60% are missense. The iminosugar 1-deoxygalactonojirimycin (DGJ, migalastat hydrochloride, AT1001) is a pharmacological chaperone that selectively binds alpha-Gal A, increasing physical stability, lysosomal trafficking, and cellular activity. To identify DGJ-responsive mutant forms of alpha-Gal A, the effect of DGJ incubation on alpha-Gal A levels was assessed in cultured lymphoblasts from males with Fabry disease representing 75 different missense mutations, one insertion, and one splice-site mutation. Baseline alpha-Gal A levels ranged from 0 to 52% of normal. Increases in alpha-Gal A levels (1.5- to 28-fold) after continuous DGJ incubation for 5 days were seen for 49 different missense mutant forms with varying EC(50) values (820 nmol/L to >1 mmol/L). Amino acid substitutions in responsive forms were located throughout both structural domains of the enzyme. Half of the missense mutant forms associated with classic (early-onset) Fabry disease and a majority (90%) associated with later-onset Fabry disease were responsive. In cultured fibroblasts from males with Fabry disease, the responses to DGJ were comparable to those of lymphoblasts with the same mutation. Importantly, elevated GL-3 levels in responsive Fabry fibroblasts were reduced after DGJ incubation, indicating that increased mutant alpha-Gal A levels can reduce accumulated substrate. These data indicate that DGJ merits further evaluation as a treatment for patients with Fabry disease with various missense mutations.

Conversion from mycophenolate mofetil to enteric-coated mycophenolate sodium in renal transplant recipients with gastrointestinal tract disorders.

OBJECTIVE: The objective of this prospective study was to evaluate the effect of conversion from mycophenolate mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) in renal transplant recipients with gastrointestinal tract (GI) reverse effects using patient-reported outcomes instrument. METHODS: A multicenter, open-label, prospective study was undertaken in renal transplant recipients treated with MMF. In patients experiencing GI tract symptoms, treatment was changed to equimolar EC-MPS (myfortic). At baseline and visit 2 (4-6 weeks after baseline), patients completed the Gastrointestinal Symptom Rating Scale (GSRS) and Gastrointestinal Quality of Life Index (GIQLI), and physicians completed the Overall Treatment Effect (OTE) scale at visit 2. A difference of 0.5 or greater in the OTE score is indicative of clinical relevance. RESULTS: Of 154 patients screened, 118 fulfilled the inclusion or exclusion criteria. Eighty-five men and 33 women with a mean age of 41.6 years participated in this study. Median time since transplantation was 12 months. Mean (SD) dose of MMF reported at baseline was 1209.4 (422.89) mg/d. More than 50% of patients reported MMF-associated nausea, dyspepsia, and abdominal pain. After conversion to an equimolar dose of EC-MPS, patients showed improvement in GI symptoms. This benefit was predominantly observed in patients with moderate to severe symptoms at baseline. On the GSRS, patients reported a significant (P < .05) reduction in symptom burden across all parameters (reflux, 36%; diarrhea, 38%; indigestion, 36%; constipation, 28%; and abdominal pain, 40%). On the GIQLI also, significant (P < .05) improvement was reported (symptoms, 18%; emotional status, 22%; physical functioning, 21%, and use of medical treatment, 18%). On the OTE scale, 84.7% of patients reported improvement in GI symptoms. CONCLUSION: In patients with moderate to severe GI symptoms, changing treatment from MMF to EC-MPS significantly reduces GI-related symptom burden and improves GI-specific quality of life.

Statistical optimization for tannase production from Aspergillus niger under submerged fermentation.

Statistically based experimental design was employed for the optimization of fermentation conditions for maximum production of enzyme tannase from Aspergillus niger. Central composite rotatable design (CCRD) falling under response surface methodology (RSM) was used. Based on the results of 'one-at-a-time' approach in submerged fermentation, the most influencing factors for tannase production from A. niger were concentrations of tannic acid and sodium nitrate, agitation rate and incubation period. Hence, to achieve the maximum yield of tannase, interaction of these factors was studied at optimum production pH of 5.0 by RSM. The optimum values of parameters obtained through RSM were 5% tannic acid, 0.8% sodium nitrate, 5.0 pH, 5 × 10(7) spores/50mL inoculum density, 150 rpm agitation and incubation period of 48 h which resulted in production of 19.7 UmL(-1) of the enzyme. This activity was almost double as compared to the amount obtained by 'one-at-a-time' approach (9.8 UmL(-1)).

A synthetic inhibitor of interleukin-1 beta converting enzyme prevents endotoxin-induced interleukin-1 beta production in vitro and in vivo.

A potent, reversible, tetrapeptide inhibitor of interleukin-1 beta converting enzyme (ICE), L-709,049, has been shown to suppress the in vitro production of mature IL-1 beta. We now report that this inhibitor also effectively suppresses the production of mature IL-1 beta in a murine model of endotoxic shock. Intraperitoneal administration of L-709,049 reduced the elevations of IL-1 beta in the plasma and peritoneal fluid of mice treated with LPS in a dose-related manner (ED50 = 2 +/- 0.9 mg/kg). LPS-induced elevations in IL-1 alpha and IL-6 in these mice were unaffected, indicating that the inhibitor specifically affected IL-1 beta production. Immunoblot analysis of plasma and peritoneal fluid indicated that L-709,049 suppressed the formation of mature IL-1 beta production in vivo. When mouse blood was incubated in vitro with LPS, IL-1 beta was released into the plasma. This assay was used to determine ex vivo the activity of an ICE inhibitor in the blood following its administration to mice. Blood obtained 15 minutes after ip administration of 10 mg/kg of L-709,049 to mice produced 80% less IL-1 beta than control blood, and IL-1 beta production returned to control levels in blood obtained 30 minutes after injection of this inhibitor. In addition, the capacity of the blood plasma obtained from these animals to prevent the cleavage of a synthetic substrate by ICE disappeared within 1 h of ip administration of 50 mg/kg of inhibitor.

Design and synthesis of potent, selective, and orally bioavailable tetrasubstituted imidazole inhibitors of p38 mitogen-activated protein kinase.

Novel potent and selective diarylimidazole inhibitors of p38 MAP (mitogen-activated protein) kinase are described which have activity in both cell-based assays of tumor necrosis factor-alpha (TNF-alpha) release and an animal model of rheumatoid arthritis. The SAR leading to the development of selectivity against c-Raf and JNK2alpha1 kinases is presented, with key features being substitution of the 4-aryl ring with m-trifluoromethyl and substitution of the 5-heteroaryl ring with a 2-amino substituent. Cell-based activity was significantly enhanced by incorporation of a 4-piperidinyl moiety at the 2-position of the imidazole which also enhanced aqueous solubility. In general, oral bioavailability of this class of compounds was found to be poor unless the imidazole was methylated on nitrogen. This work led to identification of 48, a potent (p38 MAP kinase inhibition IC50 0.24 nM) and selective p38 MAP kinase inhibitor which inhibits lipopolysaccharide-stimulated release of TNF-alpha from human blood with an IC50 2.2 nM, shows good oral bioavailability in rat and rhesus monkey, and demonstrates significant improvement in measures of disease progression in a rat adjuvant-induced arthritis model.

Role of coagulase-negative staphylococci and micrococci in ocular disease.

Forty-nine strains of coagulase-negative staphylococci and 26 strains of micrococci isolated from human eyes were classified by the Baird-Parker system of 1966. The staphylococci belonged to subgroups II, III, IV, and V with one strain each in subgroups III and IV. However, when biotyped by Baird-Parker's system of 1974, less than 50% (23/49) of strains were typable and they belonged to biotypes 1 and 2. With our proposed modified classification all strains could be classified into biotypes 1, 2, and 3. All the 26 strains of micrococci belonged to subgroup VII. Staphylococci isolated from diverse sources produced, qualitatively and quantitatively, different corneal pathology in rabbits when compared to those isolated from healthy conjunctiva. This bore no relation to the present system of subtyping based on biochemical reactions. Micrococci were incapable of producing experimental corneal lesions in the rabbit.

Determination of antimony in impure zinc sulphate solution by coprecipitation followed by differential pulse anodic stripping voltammetry.

A simple coprecipitation technique for the quantitative separation of the antimony present in impure zinc sulphate electrolyte followed by its voltammetric determination is described. Antimony in microgram levels is separated from the matrix zinc sulphate solution, which contains higher levels of copper, lead and cadmium, and is subsequently determined by differential pulse anodic stripping voltammetry (DPASV) in 3 M hydrochloric acid. Hydrous manganese dioxide is employed as the collector. This procedure, which effects considerable saving in time, is of comparable accuracy to the conventional spectrophotometric method using the antimony-rhodamine B complex. A series of synthetic zinc sulphate solutions spiked with known amounts of antimony as well as plant solutions gave near theoretical values.

Abdominal tuberculosis in children.

Problems in the management of abdominal tuberculosis in children are discussed with reference to 80 surgically proven cases. The protean clinical manifestation depends on the site and the extent of the disease and its complications. The clinical diagnosis is difficult because of the vague symptoms, non-specific signs, and non-availablity of specific diagnostic tests. The most common type of pathology seen in abdominal tuberculosis in the paediatrics age were adhesive variety followed by nodal type. Strictures of the small bowel are uncommon and hyperplastic variety is rarely seen in this age group. Response to the antitubercular drugs is excellent however, the post operative complications and mortality remain high.

Immature gastric teratoma.

Gastric teratoma, a very rare tumor of infancy is essentially benign. Immature (embryonic) elements have been described in only three of the sixty-six cases described previously. This report concerns with one "immature" gastric teratoma seen in infancy.

Inguinal herniotomy in children: a decade's experience.

We reviewed our experience with 1369 inguinal herniotomies in 1340 children performed over the last one decade. Different grades of surgeons were assigned work according to the complexity of cases. Except for the minor scrotal hematoma, other complications were hardly seen. Recurrences were seen in only 2 cases. Careful training and supervision of junior staff in the technique of inguinal herniotomy has led to results that compare favorably to those of specialized units in developed countries.

Abdominal tuberculosis in children: experience over a decade.

This presentation deals with 110 surgically proven cases of abdominal tuberculosis in the pediatric age group. The protean clinical profiles and complications of the disease entity made the clinical diagnosis difficult; the investigations were also found non-pathognomonic. The most common type of pathology seen was adhesive variety followed by nodal type. Strictures of the small bowel were uncommon and hyperplastic variety was rarely seen in the present series. The pathogenesis relating to various varieties has been suggested based on the vast experience from a single institution.

Therapeutic and diagnostic role of bronchoscopy in pediatric age group.

Two hundred and seventy five cases were evaluated bronchoscopically for various respiratory conditions. In 140 cases, a foreign body and in 30 cases mucus plug was removed. In 47 cases there was inflammation of the tracheobronchial tree. Forty patients with empyema thoracis were evaluated bronchoscopically because of persistence of bronchopleural fistula or continued pus discharge from intercostal tubes not responding to the routine treatment. The purpose of the article is to stress the therapeutic as well as diagnostic aspect of bronchoscopy for various respiratory conditions in pediatric age group.

Teratomas in pediatric age group: experience with 75 cases.

The clinicopathological features of 75 children under the age of 12 years with teratomas are reviewed. Tumors arose in the following anatomic sites: sacrococcygeum (n = 49), ovary (n = 10), Testis (n = 5), oral cavity (n = 3), retroperitoneum (n = 2) and others (n = 6). Fifty five (74%) presented within the first year of life. Excluding the gonadal tumors, male-female ratio was 2:5. Majority of the tumors had only mature tissues. Such patients and those 9 patients in whom the histology was not specified, underwent excision alone and had 95% early survival rates. Five patients had admixture of mature and immature tumors. Nine patients had malignant tissues. Germ cell tumors containing only malignant component, but no mature or immature teratomatous tissues were excluded from the series. The patients with immature and malignant tissues underwent multimodal therapy including surgical excision, multiagent chemotherapy (VAC regimen) and at times radiotherapy. Mortality in patients with immature and malignant teratomas was 20 and 66.7%, respectively. Besides histology, the only factor which affected prognosis, especially in case of sacrococcygeal teratomas was the age at the time of presentation. Our experience highlights the importance of early recognition and complete surgical excision of teratomas in the pediatric age group.

Pattern of childhood trauma. Indian perspective.

Although infectious diseases are still the chief cause of death in children in a developing country like ours, but a definite increase in incident and related mortality due to trauma has been noted in the last decade. The problems relating to pediatric trauma are peculiar to our setup and differ considerably to the severe multiple organ trauma met with in the high velocity vehicular accidents, seen in developed Western countries. The present study identifies patterns of childhood trauma from our region. It comprises 2100 patients admitted over a 3 years period to Pediatric Surgical Unit. Cranial injuries were the most commonly encountered injuries followed by the abdominal and skeletal injuries. Fall from the house roofs is the commonest mode of injury, although road traffic accidents are also recognized to be on the increase. The overall mortality was 7.7%; its chief determinant being the presence of severe head injury. A few important epidemiological factors involved were identified.

Spontaneous gastrointestinal perforation in the neonate.

Twenty seven cases of spontaneous gastro-intestinal perforation were treated from 1981 to 1990. Four perforations were in the stomach, 17 in the small bowel and seven in the large bowel. One of them had dual perforation, one in the stomach and another in the duodenum. The exact etiology remained obscure. Various factors observed were maternal obstetric complications, prematurity and perinatal asphyxia. Stress should be laid on early diagnosis by following up 'at risk neonates' thus, giving a better overall survival rate.