Preparation of prospective plant oil derived micro-emulsion vehicles for drug delivery.

Biocompatible oil-in-water (o/w) micro-emulsions can be prospective drug delivery vehicles for their capability to solubilize lipophilic (oil soluble) drugs in the dispersed oil. Plant oils are considered suitable for such a purpose. In this study, we have attempted to examine the dispersion of corn, cottonseed, clove, orange and peppermint oils, as well as isopropyl myristate (IPM) in water continuum in presence of surfactants Tween-20, Brij-30 and Brij-92 and co-surfactants ethanol (EtOH) and isopropyl alcohol (iPrOH). Both ternary (oil/surfactant/water) and psedoternary (oil/surfactant + co-surfactant/water) phase diagrams were constructed. The ternary systems produced larger micro-emulsion forming zones than the psedo-ternary systems. The combinations peppermint oil/iPrOH/water, IPM/iPrOH/water and 1:1 (v/v) peppermint oil + IPM/iPrOH/water were found to form fair proportion of single-phase surfactant-less micro-emulsion. The surfactant-aided ternary systems produced larger clear microemulsion zones, compared to pseudo-ternary systems, while the behaviour of surfactant-less systems was intermediate. The prepared systems had shelf life of 1 year and they withstood temperature variations in the range of 4-40 degrees C.

Cholera in its present day scenario.

Cholera existed in many parts of the world since olden days. Gangetic delta is considered as the home of the disease. Since 1970 there has been a significant development of the disease with its ecologic and epidemiologic aspects. Vibrio cholerae non-01 strain in taxonomically separated from V cholerae 01 strain. Though 01 strain causes epidemic outbreaks, still non-01 has been implicated to cause cholera like illness. While humans are long considered to be the only reservoir of V cholerae 01 strain, but the organism appears to have a free-living cycle in the natural environment. The organism survives more rapidly in the brackish water than fresh water. It has been demonstrated that V cholerae undergoes conversion to a viable but non-culturable state, whereby the cells are reduced in size, become ovoid, but in contrast to starved cells, do not grow at all on standard laboratory media. Seasonality coupled with starvation response and dormancy phenomenon, reflects the origin of V cholerae as an autochthonous estuary dweller. All biotypes of this organism can grow on media containing chitin as the sole carbon source. Outbreaks of the disease are related to plankton blooms associated with warmer sea-surface temperature. In 1997 V cholerae 01 biotype El tor continued to occur in all regions of the world. In 1982 a new classical variant initially displaced entrenched El tor in Bangladesh and coexisted with it for almost a decade. V cholerae 0139 Bangal has arisen along the Bay of Bengal and has spread in Asia.

Humoral immunity status in neonates born to pre-eclamptic toxaemia mothers.

A prospective study on 90 neonates born to age matched normal mothers (set I) and mothers (set II) with pre-eclamptic toxaemia (PET) was undertaken to assess and compare the humoral immunity status of the neonates. All of them had normal vaginal delivery. IgG, IgA and IgM were estimated by radial immunodiffusion technique from cord blood of neonates. It was observed that IgA and IgM levels were insignificant in the cord blood. IgG level was low in normal birth weight (NBW) neonates born to PET mothers, when compared to that of NBW neonates born to normal mothers. Again low birth weight (LBW) babies of both the sets showed lower values of IgG than that of NBW babies. Apgar scoring showed direct relationship with IgG levels e.g., higher the Apgar score higher the level of IgG. Thus the IgG level was directly related to the birth weight of the neonates of the respective sets as well as with the Apgar scoring of the neonates.

Pre-labour rupture of membrane: the histological study of membrane and bacteriological profile.

One hundred two (102) cases of pre-labour rupture of membrane (PROM) were studied and special attention was given to the histological study of the amniotic membrane as well as to the bacteriological study of high vaginal flora, cervical flora and flora of amniotic fluid, in search of probable causes or factors leading to PROM. The incidence was found to be 3.16% in the age group of 20-25 years without any relation to parity; and the duration of gestation was 38 to 40 weeks in most of the cases. The histological study revealed: (a) Focally denuded amniotic epithelium, focally separated amniotic epithelium from chorion layer, lesser density of focal squamoid change of the epithelium and thicker chorion layer probably indicating focal immaturity of the chorio-amnion, (b) lesser thickness of collagen layer, focal hydropic degeneration and mild cellular infiltrate, (c) presence of focal hyaline degeneration and focal calcification of chorio-amnion. Microbial culture revealed: (a) Higher rate of positive culture in high vaginal swab, cervical swab and amniotic fluid showing presence predominantly of Esch coli, Strept haemolyticus, klebseilla species, Staph aureus, Strept non-haemolyticus, proteus species and pseudomonas species against that of positive cultures in the control cases, (b) no anaerobic bacteria from high vaginal swab, cervical swab or from amniotic fluid. It was presumed that focal immaturity of chorio-amnion or focal irregularity in the chorio-amnion at the microscopical level, focal degeneration of collagen superadded with bacterial infection, however mild, could be the factors leading to weakness in the tensile strength of chorio-amnion, again leading to PROM, in the face of stress factors of foetal origin.

A comparative study of silver binding nucleolar organiser regions (AgNORs) of breast lesions in histological sections and fine needle aspiration smears.

The study presents a comparative profile of AgNOR dot counting in different types of breast lesions in histopathological (HP) sections and fine needle aspiration cytology (FNAC) smears. The breast lesions chosen were non-neoplastic lesion like fibroadenosis, benign neoplastic lesion like fibroadenoma and malignant neoplastic lesion like infiltrating duct carcinoma-grade 2. The AgNOR counts of non-neoplastic lesion were significantly less in number than the neoplastic lesions--both benign and malignant, in both the HP section and FNAC smear. But the counts did not show significant difference in the two neoplastic lesions eg, fibroadenoma and infiltrating duct carcinoma-grade 2, in both the HP section and FNAC smear. The appearance of the dots, as felt by the observers, were more discriminating between the three lesions, eg, uniform small compact centrally placed in fibroadenosis; mostly uniform small compact but occasional large irregular in fibroadenoma and large irregular marginally located in infiltrating duct carcinoma. Counting was easier and the appearance of the dots more easily discernible in FNAC smear than the HP section as the smear was monolayer and the malignant cells were easily detected from macrophages and stromal cells. But the tissue fluid or secretions or blood when present in the smear gave the smear a dirty background which was disturbing to the observers. Thus this AgNOR technique, when applied in HP section or FNAC smear, appears cost ineffective, lengthy and tedious procedure; did not offer absolute histochemical discriminant for malignancy from benignancy. But the shape and size distribution and appearance of the dots showing much variability in FNAC smear than the HP section, might be of some help in the diagnosis of malignancy and discriminating from benignancy.

Study of myopathies by histological and histochemical methods with special reference to staining for desmin expression.

An attempt was made to study the histological and histochemical changes as well as immunohistochemical changes in desmin expression occurring in four types of clinical myopathies e.g. Chronic ischaemic myopathy due to Buerger's disease (Group I), Carcinomatous myopathy (Group II), Metabolic myopathy (Group III) and Muscular dystrophy (Group IV). The number of cases studied were 16 cases, 15 cases, 4 cases and 5 cases respectively. The study revealed: (i) a combination of normal, degenerated, necrotic and regenerating fibres in different proportions in all the four groups having maximum number of degenerated fibres in Group I and Group IV, relatively more number of regenerating fibres in groups III and absence of necrotic fibres in Group I. (ii) Altered tinctorial property in most of the fibres indicating degenerated and regenerating fibres in all the groups with Masson's trichrome staining against inconstant staining with PTAH appear to be a good indicator for myopathy. (iii) The Desmin expression was week and irregular in most of the cases with most of the fibres probably due to reduction of desmin content probably indicating degenerated fibres, appear to be a good indicator for myopathy. (iv) Chronic ischaemic myopathy showed close resemblance with muscular dystrophy though no typical or distinct distinguishing feature could be identified in these four groups.

Cell mediated immune status in malignancy--pretherapy and post-therapy assessment.

Twenty-eight cases of malignancies of different kinds were studied to assess T-cell activity and population before and after institution of therapy. Fifteen cases were diagnosed as non-metastasising squamous cell carcinoma of larynx, pharynx, laryngopharynx, hypopharynx and tonsils. Seven cases were non-metastasising infiltrating duct carcinoma of breast and 6 cases were non-Hodgkin's lymphoma (NHL). It was observed that 3 out of 15 cases (20%) of squamous cell carcinoma cases were Mantoux test (MT) negative with a T-cell population of less than 40%, 2 out of 7 cases (28.6%) of infiltrating duct carcinoma of breast were MT negative with a T-cell population of less than 40% and 3 out of 6 cases (50%) of NHL were MT negative with a T-cell population of less than 40%. The normal controls, consisting of apparently normal healthy adults, had a T-cell population of more than 40% and were all MT positive. The patients who showed a negative skin test and a T-cell population less than 40% were further subjected to assessment of T-cell population and activity after appropriate therapy, and clinical cure of the disease. It was observed that 2 out of 3 cases (66.66%) of squamous cell carcinomas, 2 out of 2 cases (100%) of adenocarcinomas and one out of 3 cases (33.33%) of NHL showed positive conversion with a T-cell population of more than 40%.

Enterotoxicity, haemolytic activity and antibiotic susceptibility of Aeromonas eucrenophila strains isolated from water and infected fish.

Strains of A. eucrenophila isolated from fresh water (2 strains) and infected fish (4 strains) were tested for haemolytic activity and enterotoxicity and any correlation between them. Also, the resistance patterns of A. eucrenophila were tested especially in relation to ampicillin. None of the A. eucrenophila strains caused fluid accumulation in the initial tests, however, they did so only after one to four sequential passages through the gut of a susceptible host. All the strains of A. eucrenophila showed beta-haemolytic activities. Production of beta-haemolysin could be correlated with enterotoxicity. Since all the strains of A. eucrenophila were resistant to ampicillin, media containing this antibiotic may be used for their isolation from diverse sources.

Near ultraviolet radiation induced changes in goat lens.

The possibility that the ultraviolet radiation from sunlight and other ambient sources as a major causative factor for the onset of cataract processes and photolytic changes of the eye lens constituents was studied. Normal goat lenses exposed in vitro to near UV radiation in the region of 315-400 nm (UV-A) revealed distinct morphological changes in the ultrastructure, increase in the inorganic elements; C, H, N, and a sharp shift in the intrinsic fluorescence spectra. UV exposure resulted in an alteration in the lambda max of the excitation spectra, a red shift in the emission absorption maxima and also an increase in the absolute fluorescence intensity. Scanning electron microscopic study showed a significant increase in the interfibrillar distances of the lens structural proteins. It is argued that the UV light induced covalent modifications of the lens proteins and their aggregations might have occurred due to the generation of photolytic products which then lead to oxidative damages.

Isolation and characterization of the basal cell membranes of the human term placenta.

A relatively simple and rapid method is described for the isolation of basal cell membranes (BCM) from the human placenta at term which showed considerable improvement in the yield, purity and membrane characteristics as compared to the earlier described methods. The method is based on thorough washings of the syncytium in balanced salt solution, selective grinding, hypotonic lysis, sonication, incubation with EDTA and then more conventional differential centrifugation and ultracentrifugation. The isolated material showed smooth surfaced vesicular structure of various sizes as revealed by both positive and negative staining and transmission electron microscopic analysis. The membrane was highly enriched in Na+/K+, Ca2+ and Mg2+ dependent ATPase activities while the cross contamination with brush border surfaces was low as revealed by the marker enzyme assays specific for the brush border membrane (BBM) such as the disaccharide hydrolases, aminopeptidase and alkaline phosphatase. The membrane showed a relatively low lipid/protein ratio and the lipid composition represented by a variety of phospholipids (phosphatidyl choline, phosphatidyl ethanolamine, sphingomyelin, phosphatidyl inositol and phosphatidyl serine), neutral lipids (cholesterol, triacyl glycerol, free fatty acids) and glycosphingolipids (ganglioside, cerebroside and sulfatide). It also contained plasmalogens. On SDS-PAGE analysis and Coomassie blue staining reaction, the isolated membrane showed 14 major bands with as many minor ones with a molecular weight ranging between 30-110 kDa.

Ca(2+)-transport across brush border and basal surfaces of human term [correction of terms] placenta.

Brush border microvillous (BBM) and basal cell membranes (BCM) were isolated from syncytiotrophoblast of human term placenta by homogenization, sonication, prolonged stirring and differential centrifugation. Uptake of 45Ca(2+)-CaCl2 in membrane vesicles in morpholino propane sulphonic acid (MOPS) buffer was studied up to 60 min. Maximum uptake of the radioisotope was recorded at 10 and 15 min of incubation of the BBM and BCM, respectively. Radioisotopic uptake was also dependent on the Ca(2+)-concentration, linear up to 3 mu mole and then assuming hyperbolic substrate saturation kinetics. The Lineweaver-Burk transformation of the data gave Kt value for BBM and BCM, 0.85 and 1.08 microM, respectively while the Vmax of uptake (Jmax) in the same were 105.26 and 188.68 pmole Ca2+/microgram protein/20 min. Ca(2+)-Uptake in placental BBM and BCM vesicles was inhibited by two Ca(2+)-channel blockers, nifedipine and verapamil to as much as 50% while Ca(2+)-ionophore A23187 enhanced the uptake process significantly.

Fine needle aspiration cytology of ophthalmic lesions.

Forty cases of ophthalmic lesions were subjected to fine needle aspiration cytology for pre-operative diagnosis which included retinoblastomas (5 cases), inflammatory and neoplastic lesions of the eyelid and lacrimal glands (29 cases) and extra-ocular orbital lesions (6 cases) inclusive of 3 cases of rhabdomyosarcomas. The lesions were distributed in both the eyes almost in equal proportions. Histological confirmation was obtained in 85% of cases; false positive in only one case with malignant neoplasm of the eyelid and false negative also in only one case with inflammatory pseudotumour which were, histologically, proved to be adenoma of tarsal gland and lymphocytic lymphoma respectively.

Effect of Ca(2+)-channel blockers, Ca(2+)-ionophore and increased pyrene excimer formation on the microsomal glucose-6-phosphatase.

The dependence of microsomal glucose-6-phosphatase (G-6-Pase) activity on Ca2+ as well as the membrane lipid microviscosity was studied by the effect of Ca(2+)-channel blockers (namely verapamil and nifedipine), Ca(2+)-ionophore, A23187 and pyrene excimer formation. Channel blockers depressed the G-6-Pase and Ca(2+)-ATPase while the ionophore increased these activities. Dimethyl sulfoxide, a known membrane surface active agent showed no change. Ca(2+)-uptake into the membrane has expectedly been lowered by the channel blockers while the ionophores facilitated the ion flux. Excimer formation of the fluorescent probe, pyrene as an indicator of increased membrane fluidity, and microviscosity calculated from there on, showed that Ca(2+)- and lipid microenvironment in the membrane significantly influenced the activity of G-6-Pase. Membrane lipid composition such as phospholipid/cholesterol molar ratio which also indicates an increased membrane fluidity is markedly increased with the ionophore but decreased with the channel blockers, while protein/phospholipid ratio remained unchanged. Microsomal G-6-Pase is a multicomponent multifunctional protein. It is argued that Ca2+ may play the role of an obligatory cofactor not only for the hydrolysis of G-6-P (catalytic part of the enzyme) but also involved in the regulation of substrate and product transport in or out of the endoplasmic reticulum lumen.

Fine needle aspiration cytology of bone and joint lesions: an assessment.

Fine needle aspiration cytology and subsequent correlation with histology and therapeutic follow-up was done in 198 cases of bone and joint lesions. Overall accuracy was found to be 87.34%; false diagnosis was recorded in 4.5% of cases; 8.08% cases were excluded from correlation as they did not turn up for further treatment or follow-up after the diagnosis. False diagnosis was encountered mostly with inflammatory lesions and primary neoplasms, diagnosing osteosarcoma and Ewing's sarcoma as chronic inflammatory lesion, chondroma as chondrosarcoma and vice versa. The possible cause of default has been discussed. The benign cystic lesions could be diagnosed with almost 100% accuracy by fine needle aspiration cytology where 10% cases were excluded as they could not be followed up.