The unicellular glands in the epidermis of the Indian freshwater fish Mastacembelus pancalus consist of three types of mucous cells and sacciform cells. The histochemical properties of their secretory glycoproteins have been analysed by means of a battery of histochemical methods. These included methods for the identification and simultaneous visualization of oxidizable vicinal diols, O-acyl sugars, O-sulphate esters and sialic acid residues with or without side-chain O-acyl variants. Four general classes of glycoproteins (GPs) were identified. These included (i) GPs with O-sulphate esters and oxidizable vicinal diols, (ii) GPs with oxidizable vicinal diols and sialic acid residues with or without O-acyl substitution at C7, (iii) GPs mainly with O-sulphate esters, low moieties of GPs with oxidizable vicinal diols, O-acyl sugars and sialic acid residues with side-chain O-acyl variant predominantly at C8 (or which are di- or tri-substituted) or C9 and in traces of sialic acid residues without O-acyl substitution or with O-acyl substitution at C7, and (iv) GPs with traces of oxidizable vicinal diols, O-acyl sugars and sialic acid residues with O-acyl substitution at C8 (or which are di- or tri-substituted) or C9. The physiological significances of these GP classes and their release on the surface of the epidermis are discussed with special reference to their role in lubrication, protection and inhibition of the invasion and proliferation of pathogenic microorganisms in the epidermis, as adapted to the peculiar mode of life of the fish.

The purpose of this study was to examine the influence of exogenously administered melatonin on cataract formation and lipid peroxidation in newborn rats treated with buthionine sulfoximine (BSO), a drug which inhibits the rate-limiting enzyme in glutathione (GSH) synthesis, gamma-glutamylcysteine synthase, thereby depleting animals of their stores of the important intracellular antioxidant, GSH. BSO (3 mmol/kg BW) was given for three consecutive days beginning on postnatal day 2; melatonin (4 mg/kg) was injected daily beginning on postnatal day 2 and continuing until the animals were killed (either day 9 or day 17 after birth). None of the control animals (rats treated with neither BSO nor with melatonin) developed lenticular opacification during the observation period. In the BSO-treated rats, 16 of 18 animals (89%) had observable cataracts when they were examined. In rats that received both BSO and melatonin, the incidence of cataracts was highly significantly decreased, i.e., only 3 of 18 rats (7%) had observable cataracts. In addition to cataracts, the level of lipid peroxidation products (malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA)) was examined in the lens, brain, liver, lung, and kidney of control and experimental animals. In BSO-treated rats, the lens, kidney, and lung exhibited increased levels of MDA plus 4-HDA relative to those measured in the control rats; these increases were reversed in the BSO-treated rats who were injected with melatonin daily. While BSO administration did not increase basal levels of MDA plus 4-HDA in either the brain or liver, melatonin reduced levels of lipid peroxidation products below those measured in the control rats (at 17 days after birth). The changes induced by melatonin are consistent with the free-radical scavenging and antioxidative properties of this indole.

We report a method for determining haloperidol concentration in human scalp hair and discuss a possible linkage of haloperidol excretion into hair with the hair pigment melanin. First, an animal study was conducted to support the idea that hair contains amounts of haloperidol corresponding to the doses given and pigmented hair contains much more drug than does unpigmented hair. The haloperidol concentration was measured using a radioimmunoassay technique after hairs were dissolved in 2.5 N NaOH solution and the drug extracted. Pigmented and albino rats, whose hair from an area on the back had been removed beforehand by plucking, were administered either 1, 3, or 10 mg of haloperidol (i.p.) per kg body weight every day for 3 weeks. At the end of the administration period hair which had newly grown on the denuded area was plucked and collected. In each of the two groups classified by hair color the drug levels in the hair correlated with the doses given; however, the concentrations in the hair from the albino rats were much lower than those in the hair from the pigmented rats (which was less than 8.5%). Second, black and white hair was collected from each of seven human subjects with grizzled hair, who were receiving or had been administered haloperidol at fixed daily doses for more than 1 month, and the concentration of haloperidol in each type of hair was measured. In the same subject the concentration in the white hair was found to be much lower than that in the black (less than 10%).(ABSTRACT TRUNCATED AT 250 WORDS)

Summary.A The immunohistochemical methods employing colloidal gold-labeled immunoglobulins are subject to the serious limitation that immunoglobulins of all animal species will not necessarily bind firmly to colloidal gold. To circumvent such a limitation, an indirect immunohistochemical method named the protein A gold technique has been developed, in which a protein A-colloidal gold complex was used as the second stepreagent (ROMANO and ROMANO,1977; ROTH and BINDER, 1978; BENDAYAN et al., 1980; ROTH et al., 1981a, b; RoTH,1982). The protein A gold technique for immunohistology was originally developed for electron microscopy, and RoTH (1982) was the first to successfully employ this technique for light microscopy, demonstrating reddish reaction products in the rat endocrine pancreas.In the present study, attempts have been made to establish an improved method for the protein A gold technique for light microscopy, in which the contrast of the immunoreactive sites obtained by the protein A gold technique are intensified by means of our improved procedure of physical development.The method presented here has been tentatively named the protein A gold-silver staining. MATERIALS AND METHODSPieces of tissues from the splenic portion of the adult rat pancreas were fixed in Bouin's fluid for 18 hr, dehydrated in a graded ethanol series, cleared in xylene and embedded in paraffin. Sections were cut at a thickness of 4 pm, deparaffinized, hydrated and subjected to the following staining procedures. The sections were: 1) immersed for 15 min in three changes of 0.01 M phosphate buffered saline (PBS) (pH 7.4); 2) treated for 60 min with 1 % ovalbumin in PBS; 3) incubated for 60 min with guinea pig anti-porcine insulin serum (DAKO corporation, USA) diluted 1:400 with 1 °o bovine serum albumin in 0.01 M phosphate buffered saline (BSA-PBS) at room temperature in a moist chamber; 4) rinsed in four changes of PBS for 30 min; 5) treated for 15 min with loo ovalbumin in PBS; 6) incubated for 30 min with protein A gold complex preparation (E-Y Lab. Inc., USA., particle size, approximately 10 nm) diluted 40 times with BSA-PBS; 7) rinsed in four changes of 0.01 M phosphate buffer (pH 7.4) for 30 min; 8) physically developed with a solution of the formula noted below for 45-50 min at 20°C in a dark room; 9) washed in running tap water for 5 min and then immersed for 1 min in a photographic fixer diluted 5 times with water; 10) rinsed in running tap water for 10 min; 11) briefly counterstained with 0.1°o nuclear fast red in 5°o alminium sulfate aqueous solution; 12) dehydrated, cleared and mounted in a Bioleit.The formula of the developing solution used at step (8) is as follows : Solution A : 20% gum arabic aqueous solution, 45 ml; 10 silver nitrate aqueous solution, 1 ml. The gum arabic solution was prepared by centrifugation at 18,000 rpm for 30 min at 0°C and separation of the supernatant for use. Solution B comprised: distilled water, 15 ml; bromohydroquinone, 200 mg; citric acid, 300 mg. The working developing solution was prepa...

We have previously isolated from the hippocampus of young rats a novel peptide termed 'hippocampal cholinergic neurostimulating peptide' (HCNP) which specifically enhances the cholinergic activity of the septohippocampal system in vitro. Cloning and base sequence analysis of HCNP-specific cDNA from rat and human cDNA libraries revealed that the 1.1 kDa peptide aligns at the N-terminal region of its 21 kDa precursor protein. An affinity-purified rabbit antibody to rat HCNP prepared by us recognizes the C-terminal domain of the peptide, while an antibody against human HCNP binds to a large portion of the peptide. In this report we show that both antibodies react with HCNP-related components present in the soluble cytosol fraction of human brain tissue. Immunohistochemical examination of human nervous system tissues from elderly individuals revealed that Hirano bodies in Sommer's sector of the hippocampus were specifically stained by anti-HCNP antibodies. The number of HCNP-positive Hirano bodies was greater in patients with Alzheimer's disease than in normal, age-matched individuals. The immunohistochemical results were substantiated by immunoelectron microscopy. The present findings indicate that HCNP-related components accumulate in Hirano bodies, suggesting that patients who bear these inclusions may have a disturbance of the septo-hippocampal cholinergic system, considered to be of importance for learning and memory formation.