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Biomedical subjects

J Prasad

Publications and source records attributed to J Prasad.

At least 19 recordsLinked to original sources

The Clk/Sty protein kinase phosphorylates SR splicing factors and regulates their intranuclear distribution.

Mammalian Clk/Sty is the prototype for a family of dual specificity kinases (termed LAMMER kinases) that have been conserved in evolution, but whose physiological substrates are unknown. In a yeast two-hybrid screen, the Clk/Sty kinase specifically interacted with RNA binding proteins, particularly members of the serine/arginine-rich (SR) family of splicing factors. Clk/Sty itself has an serine/arginine-rich non-catalytic N-terminal region which is important for its association with SR splicing factors. In vitro, Clk/Sty efficiently phosphorylated the SR family member ASF/SF2 on serine residues located within its serine/arginine-rich region (the RS domain). Tryptic phosphopeptide mapping demonstrated that the sites on ASF/SF2 phosphorylated in vitro overlap with those phosphorylated in vivo. Immunofluorescence studies showed that a catalytically inactive form of Clk/Sty co-localized with SR proteins in nuclear speckles. Overexpression of the active Clk/Sty kinase caused a redistribution of SR proteins within the nucleus. These results suggest that Clk/Sty kinase directly regulates the activity and compartmentalization of SR splicing factors.

Amino Acid Sequence

Histogranin, a modified histone H4 fragment endowed with N-methyl-D-aspartate antagonist and immunostimulatory activities.

Histogranin is a naturally-occurring pentadecapeptide with a structure 80% homologous with that a fragment-(86-100) of histone H4. First isolated from bovine adrenal medulla, the peptide was also shown to be present in the pituitary, brain, adrenal glands, blood plasma, lungs and spleen. At the subcellular level, histogranin is concentrated in secretory vesicles and it is released from perfused bovine adrenal glands 15-35 min after stimulation with carbamylcholine as opposed to catecholamines and [Leu5]enkephalin which are released immediately after stimulation. Rat brain membranes possess specific binding sites for [125I][Ser1]histogranin with characteristics of a receptor, namely high affinity, saturability, reversibility and sensitivity to heat and proteolytic enzyme treatments. Intracerebroventricular injections of synthetic histogranin (10-100 nmol) in mice protect them against N-methyl-D-aspartate (NMDA)-induced convulsions without affecting convulsions induced by (R,S)-alpha-amino-3-hydroxy -5-methyl-4-isoxazole-propionate (AMPA), kainate and bicuculline. The peptide also binds to specific sites on human peripheral blood mononuclear cells and it evokes the release of tumor necrosis factor-alpha (TNF), interleukin-1 (IL-1) and interleukin-6 (IL-6) from isolated rat macrophages in culture. Since the structure of histone H4 is considered as one of the most conservative, it is presumed that histogranin possesses its own precursor and that its gene is distinctly expressed.

Adjuvants, Immunologic

Pilonidal disease.

All patients with pilonidal disease (abscess or sinus) managed surgically ('laying open') between 1975 and 1990 in the Colorectal Service, University Department of Surgery, Wellington School of Medicine were reviewed. A total of 323 operations (177 males, 146 females) were performed in 311 patients. Seven males and 5 females required two operations before satisfactory healing was achieved (recurrence rate 3.8%). Males were older than females (mean, 26.4 vs 21.5 years). One patient's wound bled following surgery and required immediate repacking. There were no other wound problems. One hundred and seventy-seven patients presented acutely with pilonidal abscess and 146 patients presented with pilonidal sinus. Patients with pilonidal abscess were younger than those with pilonidal sinus (male, 25.8 vs 26.9 years and female, 20.8 vs 23.5 years). There were proportionately more Maori patients who presented acutely. 'Laying open' under general anaesthesia seems to be a safe and successful method for managing pilonidal disease in all but the few patients in whom multiple operations have been performed previously or in those in whom healing has failed to occur. Based on our initial experience of 'day case' surgery the procedure could safely be done on an outpatient basis.

Abscess

Hartmann's operation: a personal experience.

This paper documents a 15 year experience with Hartmann's operation in the Colorectal Service at the Wellington School of Medicine, New Zealand. There were 31 male and 30 female patients. The majority had either complicated diverticular disease (27) or rectal cancer (27). Fifty-six patients were discharged home and five patients died within 30 days of surgery (8.2%). Of the 27 patients with complicated diverticular disease 19 proceeded to stoma closure with no mortality. Of the 27 patients who had complicated colorectal cancer only 2 had their stoma closed. There were 41 patients in whom bowel continuity was restored following construction of a Hartmann's stoma. Thirty-nine anastomoses were hand-sewn and two anastomoses were stapled. One patient developed a major anastomotic leak and one patient died postoperatively. Hartmann's operation has a definite place in the management of patients with complicated diverticular disease and recto-sigmoid cancer. The operation can be performed and the stoma closed safely in the former group but is less likely to be followed by restoration of continuity in the latter group.

Aged

Fissure in ano.

The present study documents a 15 year experience with anal dilatation in patients with fissure in ano. Patients who were unable to tolerate rectal examination were admitted urgently for anal dilatation. Patients in whom rectal examination and proctoscopy was possible were offered an anal dilator and were reviewed after 4 weeks. Patients who preferred not to use an anal dilator or who had not become asymptomatic were admitted for elective anal dilatation. Four finger anal dilatation was performed under general anaesthesia. Between 1975 and 1990 104 patients underwent 111 procedures. The male to female ratio was 1.3:1. Five patients were re-operated because of failure of resolution of symptoms. Three patients with anal fissures and Crohn's disease were successfully managed by anal dilatation. Nine patients had excision of a 'sentinel pile' in addition to anal dilatation. Ten patients were admitted acutely. One patient developed a perineal haematoma. Seventy-four procedures were performed as day cases. There was no mortality associated with the procedure. At the time of discharge from the clinic no patient complained of problems with continence. These results support our policy of gentle anal dilatation as first management choice in the treatment of anal fissure.

Adult

Interaction of histogranin and related peptides with [3H]dextromethorphan binding sites in rat brain.

Histogranin (HN) and related peptides were tested for their ability to modulate the binding of the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, [3H]dextromethorphan ([3H]DM), to rat brain membranes. HN, [Ser1]HN and the C-terminal fragment HN-(6-15) (0.1 nM-1 microM) potentiated (up to 1.6-fold) the binding of [3H]DM (5 nM) whereas the N-terminal fragment HN-(1-10) had no effect. The potentiation of [3H]DM binding by [Ser1]HN was blocked by NMDA (100 microM) and the NMDA receptor antagonist, CPP (1 microM) but not by the sigma (sigma) receptor ligand, (+)-pentazocine (0.1 microM) and the phencyclidine (PCP) receptor ligand, TCP (1 microM). Equilibrium binding experiments in presence of TCP (1 microM) to block PCP receptors indicated that [Ser1]HN (1 microM) causes a significant increase in the binding capacity (Bmax) of [3H]DM (from 2.46 to 3.46 pmol/mg protein) but no change in the apparent dissociation constant (Kd of 428 nM as compared with 487 nM). The results indicate that HN and related peptides specifically enhance the number of [3H]DM binding sites associated to the NMDA receptor complex.

Animals

The calcium binding protein of Entamoeba histolytica: expression in Escherichia coli and immunochemical characterization.

The gene encoding calcium binding protein (CaBP) of Entamoeba histolytica is single-copy and transcribes a single class of mRNA. The coding region of the gene, amplified from a genomic clone of CaBP by polymerase chain reaction, was cloned into the Escherichia coli expression vector pET-3c. The expressed protein was 15 kD in size, which agreed with the predicted size from the open reading frame. The recombinant protein showed altered mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after binding calcium and was found to be resistant to boiling, a property shared by many calcium binding proteins. A polyclonal antibody raised against the recombinant CaBP recognized a 15 kD protein in lysates from E. histolytica pathogenic strain HM-1:IMSS. The amebic protein could compete with the recombinant protein in binding to the antibody. While the antibody recognized lysates from a number of E. histolytica strains, it failed to cross-react with other Entamoeba species, e.g., E. invadens and E. moshkovskii. Moreover, no homologue of the CaBP gene could be detected in these species by using a set of degenerate primers for amplification by polymerase chain reaction (PCR). Our data demonstrate that amebic CaBP can be expressed in E. coli and that this protein is detectable only in E. histolytica but not in other Entamoeba.

Animals

Modulation of a surface antigen of Entamoeba histolytica in response to bacteria.

Changes in the cell surface of Entamoeba histolytica, a human intestinal parasite and the causative agent of amebic dysentery, were examined with a monoclonal antibody, 2D7.10, which selectively recognizes carbohydrate epitopes in some axenic amebic strains. While high-level expression of this epitope was observed in axenic amebae, it was either absent or present only in small amounts in xenic amebae. Furthermore, reassociation of the axenic amebae with intestinal flora resulted in loss of the 2D7.10 epitope. Our data suggest that surface antigens of E. histolytica can be modulated in response to bacteria and may provide an explanation for the observed influence of bacteria on amebic virulence.

Animals

Fine needle aspiration of Sister Mary Joseph's nodule.

A case of Sister Mary Joseph's nodule (umbilical metastasis) is described from a primary adenocarcinoma of the transverse colon. Needle aspiration cytology made the diagnosis which was confirmed by immunocytochemical localization of CEA, B72.3, EMA, and cytokeratin in the tumour cells. Extensive mucus production in the tumour cells was demonstrated by alcian blue and mucicarmine stains.

Abdominal Neoplasms

Lymphoid follicular hyperplasia--a distinctive feature of diversion colitis.

Diversion colitis refers to the inflammatory changes that occur in the defunctioned segment of the large intestine following diversion of the faecal stream. We report the histological features in the defunctioned rectums from seven patients: one each with severe constipation and Behçet's disease, two with Crohn's disease with rectal sparing and three with ulcerative colitis. The appearances of diversion colitis in a previously normal rectum are compared with diversion colitis with superimposed inflammatory bowel disease. Lymphoid follicular hyperplasia was found in all cases. This was marked in patients with inflammatory bowel disease, with or without initial rectal involvement. Other changes comprised surface epithelial degeneration and ulceration, mucosal inflammation including crypt abscesses, and crypt branching. Inflammatory and crypt changes were mild, except in ulcerative colitis where changes were marked and resembled those of the proximal colon. Lymphoid hyperplasia is a distinctive feature in diversion colitis. The term follicular proctitis, previously used to indicate chronic ulcerative colitis exclusively, should be re-examined.

Adult

Characterisation of ocular melanoma with cutaneous melanoma antibodies.

It can be difficult to distinguish between various forms of pale intraocular tumour, and in particular between an amelanotic malignant melanoma, a choroidal haemangioma, and a solitary metastasis. If a monoclonal antibody specific for melanoma could be identified, it might be radiolabelled to provide a scanning technique which could distinguish between an ocular melanoma and a similar lesion. This pilot, in vitro study was undertaken to determine if monoclonal antibody against cutaneous melanoma recognises any antigenic similarity in ocular melanomas. Three cutaneous melanoma MoAbs 225.28S, 376.96S, and 763-24T and a non-specific MoAb HMFG2 were studied. Cell impressions were obtained from fresh ocular malignant melanomas. Standard staining techniques with immunofluorescence were used. MoAb 225.28S, 376.96S, and 763.24T were positive in melanomas with a dominant epithelioid cell type and in those with a dominant spindle cell type. It is concluded that MoAb 225.28S, 376.96S, and 763.24T may be suitable for imaging ocular melanomas after labelling with 123I, 111In, or 99mTc.

Antibodies, Monoclonal

Perennial allergic conjunctivitis: definition, clinical characteristics and prevalence. A comparison with seasonal allergic conjunctivitis.

Perennial allergic conjunctivitis (PAC) is a perennial form of seasonal allergic (hay fever) conjunctivitis (SAC). Fourteen subjects with PAC were compared with 25 subjects with SAC. All patients met the same criteria for the diagnosis of a Type 1 ocular allergy. In a study of ophthalmic morbidity at an inner city health centre during the summer months the 3-month period prevalence of PAC was 3.5:10,000. In PAC symptoms are present throughout the year although seasonal exacerbations occurred in 79 per cent. Many similarities were found with SAC. PAC showed the same age range, length of history and symptoms as SAC although SAC was more severe. In both, conjunctival cytology showed eosinophils (PAC 43 per cent; SAC 25 per cent) and total IgE levels were raised in the tears (PAC 100 per cent; SAC 96 per cent) and serum (PAC 79 per cent; SAC 78 per cent). However, PAC differed from SAC in several respects: a history of exacerbation on exposure to house dust (PAC 42 per cent; SAC none) and an association with perennial rhinitis (PAC 75 per cent; SAC 12 per cent) were more common in PAC. The largest response on skin prick testing to a panel of antigens was to house dust mite (Dermatophagoides pteronyssinus) and this was more frequent in PAC (PAC 71 per cent; SAC 4 per cent). Dust mite antigen specific IgE was present in the tears of 78 per cent of cases of PAC, in higher levels than in serum indicating local production of IgE, whereas in SAC there were no detectable levels in any of the tears tested.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

Comparison of the ocular effects of atropine or glycopyrrolate with two I.V. induction agents.

Atropine and glycopyrrolate combined with either methohexitone or thiopentone were compared with the induction agent alone in patients undergoing electroconvulsive therapy. Patients acted as their own controls in each sub-group defined by induction agent. Pupil size, muscle twitches, presence of hiccups and the extent of oropharyngeal secretions were noted. Methohexitone produced a greater and more prolonged increase in pupil size than did thiopentone. The subsequent pupil size following the atropine-methohexitone mixture was significantly greater than that following the glycopyrrolate-methohexitone mixture. The combination of atropine with thiopentone produced a greater "secondary" mydriasis than thiopentone alone, or thiopentone combined with glycopyrrolate. The effect of the glycopyrrolate-thiopentone combination did not differ significantly from that of thiopentone alone. Methohexitone was associated with a greater incidence of hiccups, muscle twitching and excessive salivation.

Adult