Role of unstained smears in determining sample adequacy

OBJECTIVE: To determine adequacy with air-dried, unstained smears. STUDY DESIGN: The study was conducted on a total of 70 cases. The cases were divided into 2 groups. Group I consisted of all 70 and was analyzed by a consultant, observer A. Group II consisted of 41 cases from group I. In addition to observer A, a junior resident with 3 months' experience in pathology (observer B), analyzed the slides independently. The results were compared with those on stained smears. RESULTS: When correlated with stained smears, in group I, 55 of 58 (94.8%) cases were reported as adequate, and 11 of 12 cases (91.7%) were labeled inadequate. All were proven correct. In group II, stained smears confirmed that 33 of 35 (94.3%) were labeled adequate by observer A and 33 of 36 (91.7%) by observer B. Stained smears did not confirm 1 of 6 (16.6%) cases labeled inadequate by observer A and 1 of 5 (20%) cases by observer B. CONCLUSION: Unstained smear examination is an effective technique for determining adequacy. An inexperienced practitioner can perform it as well.     

Development and evaluation of a rapid flow-through immuno filtration test using recombinant filarial antigen for diagnosis of brugian and bancroftian filariasis

There is an imperative need to develop a rapid antibody test that can be used for diagnosis of clinical cases in travelers and expatriates, primary surveillance in areas of unknown endemicity, detection of early infection in childhood and for monitoring chemotherapeutic programs. A rapid-format, simple and qualitative flow through immuno filtration test has been developed for the identification of total IgG antibodies to recombinant filarial antigen WbSXP-1. This test system employs colloidal gold-protein A reagent as the antibody capture reagent. The sensitivity and specificity of the test was evaluated in a total of 1,230 serum samples. The sensitivity of the test was found to be 90.8% with brugian (n = 70) and 91.4% with bancroftian (n = 140) microfilaraemic subjects. The test showed minimum reactivity (4/10) with Loa loa microfilaria (MF) positive sera and no reactivity (0/20) with Onchocerca MF positive sera. This rapid diagnosis is found to be non-reactive with individuals having other parasitic diseases including schistosomiasis (n = 10), soil-transmitted helminthiases (n = 34) and protozoan infections (n = 33) indicating the potential of this test as a prospective method of diagnosis for both brugian and bancroftian lymphatic filariasis. Stability kinetics was studied at different temperatures and different time periods. The rapid flow-through immuno filtration test is advantageous since it can be stored at room temperature, is user friendly and is particularly applicable in the field as an initial screening method, for epidemiological monitoring of filarial infections in bancroftian and brugian endemic regions of the world.     

Bruchid (Coleoptera: Bruchidae) ovicidal phenylbutanoid from Zingiber purpureum

The larvicidal activity of the dichloromethane extract of Zingiber purpureum Roscoe (Zingiberaceae) rhizome against the second instar of Aedes aegypti (L.) (Diptera: Culicidae) is shown to be due to 4-(3',4'-dimethoxyphenyl)buta-1,3-diene. The diene also showed ovicidal activity against the bruchid Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Most of the eggs laid by bruchids on treated cowpea seeds were transparent, and very few of them contained developing embryos. The few larvae produced from these embryos were unable to penetrate the seed coat and enter the seed. Similar effects were seen when adults were exposed to the compound and then placed on untreated cowpea seeds, suggesting that a new type of maternally mediated ovicidal effect was involved. Coated and impregnated granular formulations of the extract were evaluated for use in the control of bruchid infestation of stored cowpea seeds. Coated granules showed activity similar to that of the crude extract but were found to lose activity rapidly. Impregnated granules were found to be less active than the crude extract.     

Determinants of exclusive breastfeeding in rural South India

Background

While breastfeeding rates have improved globally, disparities in breastfeeding practices persist particularly in rural and low resource settings. In India, only 56% of Indian mothers practice exclusive breastfeeding (EBF) for the recommended six months. As India leads the world in the number of preterm births, under 5 years of age malnutrition and neonatal mortality, understanding the factors associated with EBF can help improve the nutritional status for millions of infants. We assessed the factors associated with EBF in rural Mysore, India.

Methods

This cross-sectional analysis was nested within a cohort study assessing the feasibility and uptake of mobile prenatal care and HIV counseling and testing intervention in Mysore District. Multivariable logistic regression was used to identify the factors associated with EBF for infants between birth and six months. Exclusive breastfeeding was defined as breastfeeding with no other liquids or breastfeeding substitutes given to infants exceptfor medicine or oral rehydration solution, between birth and 6 months and was assessed at six months postpartum.

Results

We surveyed mothers who delivered in rural Mysore taluk between 2008 and March 2011. A total of 1292 mothers participated in the study. The overall breastfeeding rate at six months postpartum was 74.9% and the EBF rate was 48.5%. Factors associated with EBF included higher maternal age (Adjusted Odds Ratio[aOR] 1.04; 95% Confidence Interval [CI] 1.00, 1.09), lower maternal education (aOR1.56, 95% CI 1.10, 2.21), and 7–10 antenatal visits (aOR 1.57; 95% CI 1.09, 2.27). The most common reason for non-exclusive breastfeeding was the mother’s feeling that she did not have enough milk (23.7%). Infants that were not exclusively breastfed were most commonly fed formula/animal milk (42.6%) or castor oil/ghee (18.4%).

Conclusions

Less than half of the mothers in our sample reported exclusive breastfeeding in a rural region of Karnataka, India in the first six months, a rate lower than national and state level rates. Future interventions should evaluate whether antenatal education can improve breastfeeding outcomes. The only modifiable factor was number of antenatal visits. Breastfeeding education should be emphasized at every antenatal visit so that even mothers with fewer than 7–10 antenatal visits can learn the best techniques and benefits of breastfeeding.

     

An efficient method for the sequential production of lipid and carotenoids from the Chlorella Growth Factor-extracted biomass of <Emphasis Type="Italic">Chlorella vulgaris</Emphasis>

Efficient methodology for simultaneous extraction of multiple bioactive compounds from microalgae still remains a major challenge. The present study provides a method for the sequential production of three major products: Chlorella Growth Factor (CGF, a nucleotide-peptide complex enriched with vitamins, minerals, and carbohydrates), lipid, and carotenoids from Chlorella vulgaris biomass in an economically feasible manner. After protein-rich CGF was extracted, the spent biomass was found to contain 12% lipid and 3% carotenoids when extracted individually, compared to that of the un-utilized (fresh) biomass (lipid, 14%; carotenoids, 4%). When extracted simultaneously using conventional methods, the yield of lipid from “CGF and carotenoids-extracted biomass,” and carotenoids from “CGF and lipid-extracted biomass” were significantly reduced (50%). However, simultaneous extraction using different solvent mixtures such as hexane:methanol:water and pentane:methanol:water mixture-augmented lipid yield by 38.5% and carotenoids by 14%, and additionally retained chlorophyll and its derivatives. Column chromatographic approach yielded sequential production of lipid (18%), lutein (9%) with better yields as well as without chlorophyll interference. Different geometric isomers of lutein all-E-(trans)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol, 9Z(cis)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol, and 13Z(cis)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol were purified by HPLC and elucidated by CD, UV, NMR, FT-IR, and Mass spectra. In conclusion, the study provides an efficient and economically viable methodology for sequential production of lipid and lutein along with its geometrical isomers without chlorophyll influence and yield loss from the protein-rich CGF-extracted spent biomass of marine microalga, Chlorella vulgaris.     

Neurodegenerative diseases: model organisms,pathology and autophagy

A proteostasis view of neurodegeneration (ND) identifies protein aggregation as a leading causative reason for damage seen at the cellular and organ levels. While investigative therapies that aim at dissolving aggregates have failed, and the promises of silencing expression of ND associated pathogenic proteins or the deployment of engineered induced pluripotent stem cells (iPSCs) are still in the horizon, emerging literature suggests degrading aggregates through autophagy-related mechanisms hold the current potential for a possible cure. Macroautophagy (hereafter autophagy) is an intracellular degradative pathway where superfluous or unwanted cellular cargoes (such as peroxisomes, mitochondria, ribosomes, intracellular bacteria and misfolded protein aggregates) are wrapped in double membrane vesicles called autophagosomes that eventually fuses with lysosomes for their degradation. The selective branch of autophagy that deals with identification, capture and degradation of protein aggregates is called aggrephagy. Here, we cover the workings of aggrephagy detailing its selectivity towards aggregates. The diverse cellular adaptors that bridge the aggregates with the core autophagy machinery in terms of autophagosome formation are discussed. In ND, essential protein quality control mechanisms fail as the constituent components also find themselves trapped in the aggregates. Thus, although cellular aggrephagy has the potential to be upregulated, its dysfunction further aggravates the pathogenesis. This phenomenon when combined with the fact that neurons can neither dilute out the aggregates by cell division nor the dead neurons can be replaced due to low neurogenesis, makes a compelling case for aggrephagy pathway as a potential therapeutic option.