Trypanosoma cruzi: altered parasites after in vitro treatment with gangliosides, a therapeutic agent in experimental Chagas' disease.

Biochemical and structural modifications were investigated in axenic cultured Trypanosoma cruzi after treatment with gangliosides. Fluorescence anisotropy showed dose dependent increments in parasite membranes of ganglioside treated epimastigotes. NADP-GDH activity increased in parasites treated at day 4 (13%), 7 (137.2%), and 14 (28.50%) while NAD-MDH but decreased from day 7 to 21 (-5.74%, -32.22%, -27.92%). Treated parasites presented electron-lucent vacuoles opposite to the cytostoma, multilamellar bodies and dilated mitochondrion cristae, disorganized kinetoplast and altered heterochromatin structure. Gangliosides inhibited fusogenic ability (80%) and PLA2 activity (>75%) from the parasite. The same occurred with anti-PLA2 antibodies. Trypomastigotes suffered loss of cytoplasmic material and organelles when GM1 was present in culture medium. We propose that exogenous gangliosides produced: altered lipid order, inhibited membrane enzymes, the parasite energy source shifted from glucose to amino acids, ending on a structural transformation which signals parasite cell death.

Evaluation of the antiosteoporotic potential of Tinospora cordifolia in female rats.

UNLABELLED: The available courses of therapy to osteoporosis in menopausal women are limited by several side effects generated. A need therefore arises to explore herbal alternatives that are effective and safe. OBJECTIVE: Present animal studies were conducted to investigate the potential of Tinospora cordifolia (TC) ethanolic stem extract as an antiosteoporotic agent. METHODS: Three-month-old female Sprague-Dawley rats were either ovariectomized (ovx) or sham operated and treated with vehicle (benzyl benzoate:castor oil; 1:4), E(2) (1 microg/day) or TC (10, 50, 100 mg/kg b.wt) subcutaneously for 4 weeks. At the end of experiment bone mineral density of tibiae was measured by quantitative computer tomography. Serum was analyzed for the activity of alkaline phosphatase and levels of osteocalcin, cross-laps and lipids. Uterus and mammary gland were processed for histological studies. RESULTS: Ovx rats treated with TC (10 mg/kg b.wt) showed an osteoprotective effect as the bone loss in tibiae was slower than ovx controls. Serum osteocalcin and cross-laps levels were significantly reduced. All the above effects of TC were much milder than those produced by E(2). Alkaline phosphatase activity was higher in TC treatment groups. Total cholesterol and LDL levels remained unaltered but HDL levels were significantly lowered with TC (50 mg/kg b.wt) treatment. Uterus and mammary gland showed no signs of proliferation after treatment with TC extract. CONCLUSION: TC extract showed estrogen like effects in bone but not in reproductive organs like uterus and mammary gland. Thus, this study demonstrates that extract of T. cordifolia has the potential for being used as antiosteoporotic agent.

Spearmint induced hypothalamic oxidative stress and testicular anti-androgenicity in male rats - altered levels of gene expression, enzymes and hormones.

Mentha spicata Labiatae, commonly known as spearmint, can be used for various kinds of illnesses in herbal medicines and food industries. One of the prominent functions of this plant extract is its anti-androgenic activity. The present study investigated the probable correlation between oxidative stress in hypothalamic region and anti-androgenic action of this plant's aqueous extract on rats. Decreased activities of enzymes like superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase in hypothalamus of treated rats indicated spearmint induced oxidative stress. Further RT-PCR and immunoblot analysis demonstrated the decreased expression of some of the steroidogenic enzymes, cytochrome P450scc, cytochrome P450C17, 3beta-Hydroxysteroid dehydrogenase (3beta-HSD), 17beta-Hydroxysteroid dehydrogenase (17beta-HSD) and other related proteins like, steroidogenic acute regulatory protein, androgen receptor and scavenger receptor class B-1. Further, in vitro enzyme assays demonstrated depressed activities of testicular 3beta-HSD and 17beta-HSD enzymes. Histopathology indicated a decreased sperm density in cauda epididymis and degeneration of ductus deference. Our study suggested that spearmint probably induced oxidative stress in hypothalamus resulting in decreased synthesis of LH and FSH which in turn down-regulated the production of testicular testosterone through the disruption of a number of intermediate cascades.

Proceedings of resources for optimal care of acute care and emergency surgery consensus summit Donegal Ireland.

BACKGROUND: Opportunities to improve emergency surgery outcomes exist through guided better practice and reduced variability. Few attempts have been made to define optimal care in emergency surgery, and few clinically derived key performance indicators (KPIs) have been published. A summit was therefore convened to look at resources for optimal care of emergency surgery. The aim of the Donegal Summit was to set a platform in place to develop guidelines and KPIs in emergency surgery. METHODS: The project had multidisciplinary global involvement in producing consensus statements regarding emergency surgery care in key areas, and to assess feasibility of producing KPIs that could be used to monitor process and outcome of care in the future. RESULTS: Forty-four key opinion leaders in emergency surgery, across 7 disciplines from 17 countries, composed evidence-based position papers on 14 key areas of emergency surgery and 112 KPIs in 20 acute conditions or emergency systems. CONCLUSIONS: The summit was successful in achieving position papers and KPIs in emergency surgery. While position papers were limited by non-graded evidence and non-validated KPIs, the process set a foundation for the future advancement of emergency surgery.

Thyroid-epididymal relationship. I. Influence of hypothyroidism on epididymal lipids.

The influence of thyroidectomy on the lipid composition of caput and cauda epididymides have been studied. The analysis was conducted in epididymal tissues free from fluids and sperm. A general tendency towards accumulation of epididymal lipids was observed in hypothyroid rats. Hypothyroidism also brought about a differential regional response, which may be age-related. The existence of a relationship between triacylglycerols, phospholipids and diacylglycerols has been suggested. Since immediate thyroxine replacement to thyroidectomised rats maintained epididymal lipids at control levels, it is concluded that hypothyroidism has a definite influence on the epididymal lipid composition.

Thyroid epididymal relationship. II. Influence of hyperthyroidism on epididymal lipids.

The influence of experimentally induced hyperthyroidism on the lipid composition of the caput and cauda epididymides has been studied in pubertal and adult rats. Thyroxine treatment did not alter the major lipid classes in the epididymis. However, regional and age-related fluctuations in the concentration of mono-, di- and triacylglycerols have been observed. While the diacylglycerols increase, mono- and triacylglycerols were found to decrease, suggesting an inverse relationship between these fractions. Among the phospholipid fractions, phosphatidylcholine and phosphatidylethanolamine were reduced. The changes in epididymal lipid profiles give an indication that the epididymis may be yet another site responsible for fertility disturbances in hyperthyroid males. The withdrawal of thyroxine from hyperthyroid animals returned the epididymal lipid profiles to normal levels. This indicates that the effects of thyroxine in the epididymis are temporary and reversible following thyroxine withdrawal.

Phospholipase-C sensitive GPI-anchored proteins of goat sperm: possible role in sperm protection.

The role of glycosylphosphatidylinositol (GPI)-anchored sperm proteins in reproduction has been investigated. SDS-polyacrylamide gels (PAGE) analysis of goat sperm (Capra indica) indicated that several GPI-anchored proteins were released by phosphatidylinositol-specific phospholipase-C (PI-PLC) treatment. The distribution of this category of PI-PLC-sensitive GPI-anchored proteins on the surface of sperm was examined by indirect immunofluorescence. The fluorescence microscopic study clearly demonstrated that the PI-PLC-sensitive GPI-anchored proteins are confined predominantly to the head region of goat sperm. Further experiments were conducted on intact and PI-PLC treated sperm in order to decipher the function of GPI proteins. Co-incubation of sperm with peritoneal macrophages led to the enhanced phagocytosis of PI-PLC treated sperm by macrophages compared with the untreated intact sperm. Transmission electron micrographs of the macrophages acquired from the phagocytosis assay are provided to corroborate the same. From the results obtained it is inferred that one or more of the PI-PLC-sensitive GPI-anchored proteins on the sperm surface could act as protection factor(s) that shield the sperm from macrophages.

Anti-inflammatory and immunologically active polysaccharides of Periandra mediterranea.

Three polysaccharides, glucans with mean M(r)'s of 1.5 x 10(5), 3.6 x 10(4) and 2.1 x 10(4), were isolated from dried roots of Periandra mediterranea by fractionation on Sephacryl S-300 HR and Sephadex G-25. Chemical and spectroscopic studies indicated that they have a highly branched glucan type structure composed of alpha-(1-->4) linked D-glucopyranose residues with both (3-->4) and (4-->6) branching points. The polysaccharides enhance phagocytosis in vivo, and exhibit anti-inflammatory activity.

Plant natural products active against snake bite--the molecular approach.

The article surveys the substances identified in plants reputed to neutralize the effects of snake venoms. Protective activity of many of them against the lethal action of the venom of the jararaca (Bothrops jararaca) snake was confirmed by biological assays. It was shown that all belong to chemical classes capable of interacting with macromolecular targets--receptors and enzymes. In a few cases it has been shown that exogenous natural micromolecules can mimic the biological activity of endogenous macromolecules. From the evidence presented, it can be inferred that micromolecules which neutralize the action of snake venoms mechanistically replace endogenous antitoxic serum proteins with venom neutralizing capacity such as produced by some animals.

Effect of treatment with fluosol and mannitol during temporary middle cerebral artery occlusion in cats.

Four groups of seven cats each were subjected to 6 hours of temporary middle cerebral artery (MCA) occlusion followed by reperfusion. During the first 30 minutes of the occlusion, Group 1 received 20% Fluosol-DA (FDA) and Group 2 received 20% FDA and mannitol; Group 3 received 20% FDA and mannitol 30 minutes before the occlusion, and Group 4 (control) received an infusion of saline solution during the first 30 minutes of occlusion. All cats breathed 100% oxygen throughout the occlusion and during 2 hours of reperfusion. Blood pressure, PaO2, and pH remained stable for 8 hours. PaO2 increased from an average base line level of 102 +/- 15 (SD) torr to 520 +/- 45 torr in Group 1, 450 +/- 41 torr in Group 2, 480 +/- 28 torr in Group 3, and 396 +/- 151 torr in Group 4 during the administration of 100% oxygen. The average fluorocrit during occlusion was 6% in Groups 1, 2, and 3. The 24-hour mortality rate was 71% in Group 1, 43% in Group 2, 29% in Group 3, and 14% in Group 4; a midline shift and herniation were present after each fatal infarct. The survivors were sacrificed 10 days after the occlusion. There was a trend toward improvement of neurological function at sacrifice in Groups 2 and 3 compared with Groups 1 and 4. The infarct areas on coronal sections were 2.4 times larger in Group 1 (FDA only) than in Group 4 (control) (P less than 0.05). There was no significant difference in infarct size between Group 4 and Group 2 or 3.(ABSTRACT TRUNCATED AT 250 WORDS)

A comprehensive evaluation of the reproductive toxicity of Quassia amara in male rats.

Chloroform extracts of the bark of Quassia amara in different dilutions was used to assess its impact on the male reproductive system of albino rats. Single daily intramuscular injections of the extract for 15 days resulted in a significant reduction in the weight of testis and epididymis but not that of the seminal vesicles and prostate (all lobes). A marked decrease in the sperm count, motility, viability was also observed in sperm collected from the cauda epididymis of treated animals. A number of abnormalities like double heads, double tails, detached heads and fragile tails were frequently seen. Epididymal alpha-glucosidase activity was drastically reduced. However, prostatic acid phosphatase activity and citric acid levels and seminal vesicle fructose concentrations remained unchanged following treatment. Thus, it appears that the prime site of action is at the level of both the testis and the epididymis. Examination of the blood showed that cell counts and hemoglobin levels were in the normal range. Bilirubin, SGPT, SGOT, protein and urea were also not altered by the herbal extract. From the selective action on the male reproductive tract we suggest that the chloroform extract of the bark of Quassia amara has potential for use as an antifertility agent.

Inductively coupled plasma emission spectroscopic and flame photometric analysis of goat epididymal fluid.

AIM: The elemental composition of the epididymal luminal fluid (ELF) in adult goat (Capra indica) was investigated. METHODS: ELF was collected by micropuncture from twelve sites along the epididymal duct. The elemental contents was analyzed with inductively coupled plasma (ICP) emission spectroscopy, a microanalytical technique that can simultaneously measure many elements in minute volumes of sample. The Na and K concentrations were determined by flame photometry. RESULTS: ICP spectroscopy showed the presence of copper, calcium, nickel, iron, magnesium, chromium, titanium and zinc in ELF, with fluctuating levels at different sites along the length of the epididymis. Cadmium, cobalt, lead and manganese were not found. The Na+/K+ ratio was seen to be higher at the initial segments of the epididymis and lower at the distal. CONCLUSION: It is proposed that the observed characteristic distribution of elements in ELF may have far reaching implications in sperm maturation and storage known to occur in the epididymis.

N-acetyl beta-D-glucosaminidase is not attached to human sperm membranes through the glycosylphosphatidyl inositol (GPI)-anchor.

AIM: The mode of anchorage of N-acetyl beta-D-glucosaminidase (NAGA) on human ejaculated sperm was investigated. METHODS: Sperm plasma membrane was prepared by discontinuous sucrose gradient centrifugation from human sperm. NAGA was solubilized from these membranes by two detergents: octyl-glycoside and triton X-100. In separate studies, the release of the enzyme from the sperm membrane preparation by phosphatidylinositol specific phospholipase C (PI-PLC) was also examined. RESULTS: NAGA activity was detected on sperm membranes isolated from human ejaculates. The pattern of the enzyme solubilization by detergents indicated that the enzyme was an integral protein of sperm membrane. NAGA was not released from the sperm membranes by PI-PLC treatment. CONCLUSION: The evidence presented strongly suggests that human sperm membrane bound NAGA is not attached via the GPI anchor.

Ganoderma lucidum: partial characterization of spore and whole body antigenic extracts.

This study focused on the characterization of antigenic/ allergenic profiles of Ganoderma lucidum spore and whole body preparations. Whole body G. lucidum contained higher protein to carbohydrate ratio whereas it was less than one for spore extract. Isoelectric focusing showed 12 and 11 bands in acidic pH range (pI 3.5-6.5) for G. lucidum spore and whole body, respectively, while SDS-PAGE showed 8 and 23 fractions, respectively, in molecular weight range of 12.8-75.0 kD. The prominent protein fractions of G. lucidum spores were 19.4, 22.8 and 23.8 kD, whereas for G. lucidum whole body, 13.2, 14.7, 18. 7, 21.5 and 23.5 constituted major fractions. Immunoblotting with 41 individual serum samples revealed 21.8, 23.8, 19.4 and 20.0 kD to be major allergenic protein fractions of G. lucidum spores. The same using G. lucidum whole body and 26 individual serum samples identified several fractions of 17.0, 17.5, 18.5, 22.0, 23.8, 42.0, 44.0, 56.0 and 69.0 kD as major allergens. The compiled data suggest that there are common as well as specific allergenic components in two G. lucidum extracts studied.

Effect of blood flow rate and donor vessel diameter on the patency of carotid venous bypass grafts in dogs.

A saphenous vein graft was implanted from the right subclavian to the right common carotid artery in seven dogs (group I) and between the right and left common carotid arteries in another seven dogs (group II). The recipient artery was ligated proximally to augment blood flow through the graft. Immediately after the anastomoses were completed, the average blood flow through the graft was 32 +/- 25 mL/min in group I and 122 +/- 22 mL/min in group II. At sacrifice 30 days later, angiography showed that all grafts in group I were thrombosed, whereas six (87.5%) of seven grafts in group II were patent. These findings suggest that a larger donor vessel diameter and higher graft flow rates may improve patency in venous bypass grafts that are 3 to 5 mm in diameter.

Survey of airborne culturable and non-culturable fungi at different sites in Delhi metropolis.

A two year aerobiological survey for culturable and non-culturable fungi was conducted at human height at five different sites in Delhi metropolis. Burkard Personal Volumetric Sampler for petriplates and slide exposures were used for sampling the air. With simultaneous petriplate and slide exposure a total of 98 fungal forms were recorded. Cladosporium contributed for 25-40% of total airborne fungi followed by Ustilago (smuts) (24%) Aspergillus flavus (10-13%), Alternaria (11%) and A. niger (8%). Basidiomycetes contributed 7-13% at different sites. The frequency of occurrence of these types varied from 50-98%. In general fungal concentration was high from July to April with low counts in winter (January) and dry and hot summer (May-June). Quantitative variations in the spore counts were found to be statistically significant within the same urban locality.

Fomes pectinatis: an aeroallergen in India.

This work is focused on the aerobiology and allergenicity of Fomes pectinatis in India. The atmospheric concentration of Fomes basidiospores was recorded and the antigens were prepared from spore (FSE) and whole body (FWBE) materials. The intradermal (ID) and prick (PT) skin tests were conducted on 172 patients having respiratory allergy. The period from July to October has been recorded as having a higher concentration of Fomes spores. The maximum counts (67 spores/m3) were observed from the North Delhi site in the month of July, 1989, compared with 550 spores/m3 in the South Delhi site. Marked skin positivity (2+ and above) varied from 9.8% to FSE to 22% to FWBE. Nine out of twelve ID positive patients (2+ to 3+) to FSE also gave PT positive response. For FWBE, similar ID and PT response was obtained in 80% of cases. The soluble protein content of FSE was 0.37 mg/ml, whereas, for FWBE it was 0.70 mg/ml. It was observed through ELISA that almost all patients had significantly raised FP specific IgE levels in their sera. The current study, therefore, indicates that Fomes pectinatis may be a prevalent aeroallergen in India.

Modulation in activity of some epididymal glycosidases by prolactin.

Activity of glycosidases in the epididymis was influenced by several factors originating in the testis. Activities of all the three glycosidases studied viz., beta-glucosidase, beta-galactosidase and alpha-mannosidase were found to be significantly lower in the epididymis of orchidectomized animals than in sham operated rats. However, an enhanced activity of epididymal beta-galactosidase and alpha-mannosidase was noticed in prolactin treated orchidectomized rats compared to orchidectomized rats given vehicle alone. On the other hand, activity of these two enzymes in bromocriptine treated orchidectomized rats was even lower than that found in orchidectomized rats given vehicle. Neither prolactin nor bromocriptine treatment had any significant influence on the epididymal beta-glucosidase. The results suggest a selective but definite action of prolactin on epididymal glycosidases.

Impact of prolactin on epididymal lipid profile in castrated rats.

Prolactin treatment to castrated rats led to accumulation of triacylglycerol and esterified cholesterol. There was no appreciable drift in epididymal cholesterol: phospholipid ratio between the prolactin treated and control animals. However, further analysis of phospholipids showed a build up of phosphatidyl inositol, phosphatidyl choline and phosphatidyl ethanolamine but a drop in the levels of phosphatidyl serine and sphingomyelin in prolactin treated castrated rats as compared to those castrated animals injected with vehicle alone. Changes in phospholipids reported above were prominently seen in the group of castrated rats that received 100 micrograms oPRL/100 g body weight but not in those animals which received either lower or higher doses of the hormone. Interestingly, bromocryptine treatment in castrated rats produced a general depletion in the levels of all lipid classes studied in the epididymis. It is suggested that this may be due to impaired synthesis and/or increased breakdown of lipids in this organ.