Five groups of subjects underwent EEG sleep recordings, arecoline rapid eye movement (REM) induction response testing, and Schedule for Affective Disorders and Schizophrenia (SADS) interview. Group I: 20 patients with primary major depressive disorder (MDD) (endogenous) without any coexisting anxiety disorder; Group II: 19 primary MDD (endogenous) patients with secondary panic, GAD, or phobic disorders; Group III: 18 patients with primary anxiety disorder without coexisting MDD; Group IV: 14 patients with primary anxiety plus secondary MDD; Group V: 26 normal controls. Modified Research Diagnostic Criteria (RDC) were used for diagnosis, based on the SADS interview. There was considerable overlap of SADS scaled scores between patient groups, which is consistent with a heterogeneous clinical presentation of depressive and anxiety states. REM latency was significantly shorter in patients with primary MDD (without anxiety) as compared with that in patients with primary anxiety (no MDD) and normals. Arecoline REM induction response time was significantly shorter in both primary affective groups (I and II) as compared with primary anxiety (no MDD) patients and normal controls. REM latency and arecoline REM induction time was not significantly different between the primary anxiety groups (III and IV) and normals. The study highlights the use of biological markers in differentiating between clinical syndromes confounded by mixed or overlapping phenomenology.

Adult↗

PubMed1985

Plasmodium gallinaceum: critical role for microtubules in the transformation of zygotes into Ookinetes.

The role of microtubules and microfilaments in the transformation of spherical zygotes of Plasmodium gallinaceum (avian malaria parasite) into vermiform ookinetes has been studied by using specific drugs (taxol, colchicine, and cytochalasin-B). Both taxol and colchicine completely abolished the transformation of zygotes into ookinetes. The inhibitory effect was seen only if the drugs were added during the initial 6 hr of total time (20-24 hr) required for complete transformation; the addition of drugs after 6-8 hr of initiation of transformation had no effect. Electron microscopy revealed that microtubules were depolymerized by colchicine treatment, whereas in taxol-treated cells there was an extensive array of cytoplasmic and nuclear microtubules which appeared to be clumped in bundles. In contrast to the effects of taxol and colchicine, cytochalasin-B, which affects the microfilament system, had no effect on the transformation. Protein synthesis and expression of two ookinete-specific surface proteins were not affected in the drug-inhibited parasites. Zygotes treated with taxol for 4 hr at room temperature failed to develop into oocysts when they were subsequently fed to mosquitoes. These studies demonstrate a critical role for microtubules in the initial stages of transformation of zygotes into ookinetes.

Alkaloids↗

PubMed1985

A method for the controlled cleavage of disulfide bonds in proteins in the absence of denaturants.

A simple method was developed for the controlled cleavage of protein disulfide bonds and the simultaneous blockage of the free sulfhydryl groups in the absence of a denaturant. The disulfide bonds of bovine serum albumin were cleaved unsymmetrically at pH 7.0 using 0.1 M sulfite in 0.1 M phosphate buffer and the free sulfhydryl groups formed were sulfonated in an oxidation-reduction cycle using molecular oxygen and 400 microM cupric sulfate as a catalyst. The reaction was affected by cupric ion concentration, sulfite concentration, reaction pH and temperature. The standardized method was successfully used to cleave the disulfide bonds of other proteins pepsin, trypsin, and chymotrypsin. The method is reliable and can be used for achieving progressive cleavage of disulfide bonds in proteins without employing a denaturant.

Animals↗

PubMed1985

Phase separation in Triton X-114 of antigens of transmission blocking immunity in Plasmodium gallinaceum.

The distribution of proteins of mosquito midgut forms of Plasmodium gallinaceum in the detergent-free (aqueous) and detergent-enriched phases was studied using a phase separation technique in Triton X-114. Of the three surface proteins on gametes and newly fertilized zygotes (240, 56, and 54 kDa) immunoprecipitated by transmission blocking monoclonal antibodies, 240 kDa protein was recovered in the aqueous phase, whereas 56 and 54 kDa proteins were found preferentially in the detergent phase. The hydrophobic properties of the 56 and 54 kDa proteins were also shown by their strong tendency to interact with the lipid bilayers and a hydrophobic matrix phenyl-Sepharose. Monoclonal antibody IID3B3 immunoprecipitated all the three proteins from the whole Triton extract but in the phase-separated extracts reacted only with the 240 kDa protein in the aqueous phase and not with the 56 and 54 kDa doublet in the detergent phase. In Western blot analysis also monoclonal antibody IID3B3 reacted only with the 240 kDa protein. The 240 kDa protein in the aqueous phase was retained by monoclonal antibody IID3B3 linked to Sepharose 4B beads and could be eluted either with 0.1 M acetic acid or 50 mM diethylamine. The 56 and 54 kDa doublet in the detergent phase could be bound to and eluted from Sepharose 4B beads-linked monoclonal antibody IID4 or rabbit anti-male P. gallinaceum gamete serum. Two stage-specific glycoproteins of 26 and 28 kDa on the surface of ookinetes of P. gallinaceum were also separated in the detergent phase following Triton X-114 extraction. Phase separation in Triton X-114 offers a simple approach to the separation of a select group of proteins from the bulk of the cellular proteins.

Animals↗

PubMed1985

Biosynthesis of two stage-specific membrane proteins during transformation of Plasmodium gallinaceum zygotes into ookinetes.

We have studied the synthesis and expression of surface proteins in zygotes of Plasmodium gallinaceum during their transformation to mature ookinetes. The cells were biosynthetically labelled in vitro using [35S]methionine and proteins were immunoprecipitated with rabbit anti-ookinete serum or monoclonal antibodies. Early zygotes (approx. 2 h post-gametogenesis and fertilization) synthesized and expressed on their surface a protein of Mr 26 000 as observed under reducing conditions on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) (31 000 under non-reducing conditions) and continued to do so for 8-10 h; thereafter synthesis of the Mr 26 000 protein declined and little or none was synthesized in the mature ookinetes (greater than 20 h post-gametogenesis). Between 3-5 h post-gametogenesis, zygotes also began to synthesize a protein of Mr 28 000 (34 000 under non-reducing conditions). Synthesis and expression of this surface protein continued throughout development; and the Mr 28 000 protein was the predominant surface protein synthesized by the mature ookinete. Mr 26 000 and Mr 28 000 proteins have been designated earlier as PgO-1 and PgO-2 respectively (Carter and Kaushal, Mol. Biochem. Parasitol. (1984) 13, 235-241). Neither protein was synthesized in the gametocytes prior to gametogenesis. Both proteins could be labelled with [3H]glucosamine or [3H]mannose. When zygotes were incubated with [3H]palmitic acid both PgO-1 and PgO-2 bound fatty acids in covalent linkage. The two proteins do not otherwise appear to be structurally related. They were differentially immunoprecipitated by different monoclonal antibodies and gave rise to distinct patterns of peptides following digestion with proteases such as Staphylococcus aureus V-8, trypsin and chymotrypsin.

Animals↗

PubMed1985

Repeated endoscopic sclerotherapy for active variceal bleeding.

Emergency endoscopic sclerotherapy (EEST) during continued moderate to severe bleeding was carried out using a flexible endoscope and absolute alcohol as a sclerosant in 38 patients with variceal bleeding. Portal hypertension was due to cirrhosis in 27, noncirrhotic portal fibrosis in eight, extrahepatic obstruction in two, and Budd-Chiari Syndrome in one patient. A technically adequate EEST could be carried out in 36 (95%) patients, with successful control of variceal bleeding in 35 (92%). Thirty-one episodes of rebleeding occurred in 12 (31.6%) patients. Twenty-nine (93.5%) of these episodes could be controlled with repeated EEST, giving an overall success of 87%. The new approach of repeating sclerotherapy on every rebleeding episode up to a maximum of three course within 24 hours, use of a wide bore injector, and certain other technical innovations were found safe and effective. The mean (+/- SD) amount of alcohol injected per patient was 9.23 +/- 3.3 ml and the mean (+/- SD) number of injections needed per patient were 6.0 +/- 3.07. Complications were minor, transient, and similar to conventional sclerotherapy. There were three deaths, two due to massive rebleeding and one due to hepatic encephalopathy. It can be concluded that EEST is technically feasible during active variceal bleeding and is an effective and relatively safe procedure. It can serve as the first line treatment in this group of patients.

Adolescent↗