Prevalence of Stigma Among TB Patients and Its Associated Factors - A Community Based Cross-Sectional Study in Puducherry, India

Background: Tuberculosis (TB) is a disease that has both medical as well as social dimensions. Stigma and associated discrimination experienced by persons affected by TB is a barrier to the interventions towards TB elimination. A better understanding of stigma will improve the effectiveness of the interventions aimed to al-leviate the effects of TB stigma. The objectives were to measure the prevalence of self, anticipated, experi-enced stigma and its associated factors and to explore the impact of tuberculosis related stigma among TB pa-tients. Methodology: A cross-sectional study was conducted in Puducherry district, Southern India among 420 adult drug sensitive non-HIV TB patients registered under NTEP. Data was collected by face-to-face interview using standardised questionnaire. Results: The mean(±SD) age of the study participants was 44.5 (±15.03) years. Majority were males (267, 63.6%) and were married (330, 78.6%). The prevalence of stigma among TB patients was found to be 69.3%. Perceived stigma was noted in 47.1%, 33.6% had self- stigma and 26.0% had experienced stigma. On measur-ing the impact of stigma, 52.6% reported participation restriction. Illiteracy and lower-socio economic status were found to be significantly associated with TB stigma. Conclusions: Stigma among TB patients was high. Tailored patient-centric interventions to address stigma and discrimination using culturally appropriate and locally available resources are necessary

A retrospective study on timeliness of vaccination among children aged 0 to 23 months in a rural area of Pondicherry

Background: Immunization is the cost-effective public health intervention that prevents and protects against vaccine preventable diseases. The objective was to estimate the timeliness in receiving age appropriate vaccines and to study selected factors influencing the timeliness of age appropriate vaccines as per national immunization schedule among children aged 0 to 23 months in a rural area of Pondicherry. Methods: A retrospective study was done at a Community Health Centre, Karikalampakkam, Pondicherry using data from immunization registers of children aged 0 to 23 months, who were born between July 01, 2013 to July 31, 2015. If the child was vaccinated within 7 days of the scheduled time, it was considered as timely vaccination. Results: Out of 679 children, 52% were males and 48% were females. The median days of delay in vaccination were ranged from 1-171 days. The proportion and the median days of delay were increased progressively as the age of the child increased. The place of delivery was significantly associated with birth doses of OPV, Hepatitis B and BCG vaccination. There was a significant difference in timeliness of vaccination across the birth order of the children for the first, second and third doses of OPV and Pentavalent vaccines (p=0.02). Birth weight of the children was not statistically associated with vaccination delay. Conclusions: Delay in vaccination in varying frequency was observed for the vaccines administered under the national immunisation schedule. Hence, the age-appropriate vaccinations should be given up-to-date as well as on time.

A cross sectional study on optimal infant and young child feeding practices with reference to WHO indicators in a rural area of Puducherry

Background: Optimal infant and young child feeding practices (IYCF) are essential to address the increasing burden of malnutrition and for the overall development of the children. The present study was conducted to estimate the proportion of optimal infant and young child feeding practices among rural children aged 0 to 23 months and study the associated socio demographic factors.Methods: A community based cross-sectional study was conducted among 360 children in the age group of 0 to 23 months in a rural field practice area of a medical college in Puducherry. Data on IYCF practices were collected using a standardized tool developed by WHO. Core and optional IYCF indicators were calculated. Chi-square test and Fishers’ exact test were used as tests of significance.Results: Almost 88.0% of infants were initiated early on breastfeeding and 90.8% were exclusively breastfed for six months. Children who were continuously breastfed at one year and two years were 77.4% and 22.4% respectively. About 75.0% were introduced with solid or semisolid foods at 6 to 8 months of age. Among children aged 6 to 23 months, 77.3% had the recommended minimum dietary diversity, 81.3% had the minimum meal frequencies, while 57.7% received the minimum acceptable diet and only 39.4% consumed iron rich foods. Gender was significantly associated with the practice of continued breastfeeding at one year, adequate minimum dietary diversity and minimum meal frequency.Conclusions: The core and optional IYCF indicators were acceptably good in the initial six months of life but thereafter showed suboptimal levels, which should be emphasized among the mothers or primary care givers.

A cross sectional study on screening for depression among elderly in rural areas of Puducherry, India

Background: Depression among elderly has been a neglected health issue. Various factors hinder its early diagnosis. The objectives of this study are to screen for depression among elderly using geriatric depression scale-15 and to determine its influencing factors.Methods: A community based cross sectional study on geriatric depression was done in villages catered by a primary health centre in Puducherry. Permanent residents of the study setting, aged ≥60 years consenting for the study were included. Line listing of elderly individuals in the study setting was done and subjects were selected by simple random sampling. A pretested structured questionnaire was used to collect socio-demographic and clinic-psychological variables. Hindi mental state examination scale was used to screen for cognitive impairment (score <23). Geriatric Depression Scale (GDS-15) was used to screen for depression (score >5) among geriatric individuals. Appropriate statistical tests of significance were done.Results: Among the 360 elderly individuals screened using GDS-15 tool, 41.4% of them were found to be at a higher risk of developing depression. Significant factors for risk of depression in univariate analysis include being female, illiterate, unemployed, widow/single, having sleep problems, dependency in activities of daily living and cognitive impairment. Predictors for risk of depression evident by multivariate analysis include presence of unemployment, sleep problems and cognitive impairment.Conclusions: A higher proportion of elderly was observed to be at risk of depression.  Primary care physicians may consider screening for depression in elderly with identified risk factors and act accordingly.

Trichoderma harzianum MTCC 5179 impacts the population and functional dynamics of microbial community in the rhizosphere of black pepper (Piper nigrum L )

Abstract Employing Illumina Hiseq whole genome metagenome sequencing approach, we studied the impact of Trichoderma harzianum on altering the microbial community and its functional dynamics in the rhizhosphere soil of black pepper (Piper nigrum L.). The metagenomic datasets from the rhizosphere with (treatment) and without (control) T. harzianum inoculation were annotated using dual approach, i.e., stand alone and MG-RAST. The probiotic application of T. harzianum in the rhizhosphere soil of black pepper impacted the population dynamics of rhizosphere bacteria, archae, eukaryote as reflected through the selective recruitment of bacteria [Acidobacteriaceae bacterium (p = 1.24e-12), Candidatus koribacter versatilis (p = 2.66e-10)] and fungi [(Fusarium oxysporum (p = 0.013), Talaromyces stipitatus (p = 0.219) and Pestalotiopsis fici (p = 0.443)] in terms of abundance in population and bacterial chemotaxis (p = 0.012), iron metabolism (p = 2.97e-5) with the reduction in abundance for pathogenicity islands (p = 7.30e-3), phages and prophages (p = 7.30e-3) with regard to functional abundance. Interestingly, it was found that the enriched functional metagenomic signatures on phytoremediation such as benzoate transport and degradation (p = 2.34e-4), and degradation of heterocyclic aromatic compounds (p = 3.59e-13) in the treatment influenced the rhizosphere micro ecosystem favoring growth and health of pepper plant. The population dynamics and functional richness of rhizosphere ecosystem in black pepper influenced by the treatment with T. harzianum provides the ecological importance of T. harzianum in the cultivation of black pepper.

Influence of temperature on symptom expression, detection of host factors in virus infected Piper nigrum L.

Expression of symptoms in black pepper plants (Piper nigrum) infected with Piper yellow mottle virus (PYMoV) vary depending on the season, being high during summer months. Here, we explored the influence of temperature on symptom expression in PYMoV infected P. nigrum. Our controlled environment study revealed increase in virus titer, total proteins, IAA and reducing sugars when exposed to temperature stress. There was change in the 2-D separated protein before and after exposure. The 2-D proteomics LC-MS identified host and viral proteins suggesting virus-host interaction during symptom expression. The analysis as well as detection of host biochemical compounds may help in understanding the detailed mechanisms underlying the viral replication and damage to the crop, and thereby plan management strategies.

Rotary endodontics in primary teeth - a review

Endodontic treatment in primary teeth can be challenging and time consuming, especially during canal preparation, which is considered one of the most important steps in root canal therapy. The conventional instrumentation technique for primary teeth remains the "gold-standard" over hand instrumentation, which makes procedures much more time consuming and adversely affects both clinicians and patients. Recently nickel-titanium [Ni-Ti] rotary files have been developed for use in pediatric endodontics. Using rotary instruments for primary tooth pulpectomies is cost effective and results in fills that are consistently uniform and predictable. This article reviews the use of nickel-titanium rotary files as root canal instrumentation in primary teeth. The pulpectomy technique is described here according to different authors and the advantages and disadvantages of using rotary files are discussed

An indirect ELISA for the diagnosis of melioidosis.

BACKGROUND & OBJECTIVES: Very little information is available on melioidosis in India. This disease caused by Burkholderia pseudomallei is not often considered as a differential diagnosis and patients are not usually investigated for it. Thus we are unaware of its prevalence in India. This study was undertaken to detect the presence of melioidosis in patients presenting with pyrexia of unknown origin (PUO) using an indirect ELISA. METHODS: The well established ELISA technique was adapted to detect melioidosis in patients attending the Christian Medical College and Hospital, Vellore and to provide a serological test using reagents with a reasonable shelf-life. The ELISA is designed to detect IgG antibodies to B. pseudomallei in serum samples. RESULTS: A cut-off optical density (OD) of 0.36 (mean +/- 2 SD of healthy controls) was chosen as diagnostic criterion for the diseased group. The mean OD values in the sera of patients with culture proven melioidosis was significantly (P < 0.001) higher than that of healthy controls. INTERPRETATION & CONCLUSION: The indirect ELISA was simple to perform and may be recommended as a diagnostic serological test when melioidosis is considered as a differential diagnosis.

Status of Calpain In gene deleted carriers of Duchenne Muscular Dystrophy (DMD).

Though DNA testing gives precise diagnosis, molecular heterogeneity of disease makes testing an elaborate effort in Duchenne Muscular Dystrophy (DMD) in spite of the fact that 60°% of the cases are caused by deletions. The molecular diagnosis of carriers is elaborate needing quantitation of amplifed exonic products. Use of polymorphic markers for other mutations are also involved. We had reported in a large pedigree of DMD with one exonic deletion, the status of calpain by ELISA, CPK and deletion assessed by quantitative PCR (QPCR) in mothers and female siblings. The results pointed to the fact that calpain test is positive in true carriers (showing gene deletion). Status of catpain was assessed in many unrelated female members of DMD families carrying other exonic deletions, which was positive to calpain changes in true carriers.

Bacteremia due to beta haemolytic streptococci.

The Beta haemloytic streptococci (BHS) are well recognised human pathogens causing a variety of infections, including septicemia. It is important to ensure their isolation from clinical specimens by using optimum media. Moreover, since the different groups have different pathogenic potential, it is equally important to routinely serogroup them; this is emphasized here. Since, BHS are uniformly will greatly decrease morbidity and mortality due to BHS infection.

Deletion analysis & calpain status for carrier detection in a family with Duchenne muscular dystrophy.

Eight females with a family history of Duchenne muscular dystrophy (DMD) were analysed for their carrier status by m-calpain test, which monitors the m-calpain (milli-calpain), a proteolytic enzyme in the platelets, using an ELISA technique. Four of the eight females were identified as carriers by virtue of their elevated enzyme levels as compared to control. DNA samples of these members were analysed to ascertain the carrier status, by PCR followed by dosage analysis by densitometry. DNA analysis confirmed the findings by calpain test, which underlines the reliability of this phenotypic test for carrier detection in DMD. Calpain test has been informative in a large group of patients and carriers tested so far. Since the calpain test is cost and labour effective, it is suited for routine and widespread screening purposes.

Incidence of methicillin resistant coagulase positive & coagulase negative staphylococci in blood cultures.

Isolates of Staphylococcus aureus and coagulase negative staphylococci (CONS) from blood culture of bacteraemic patients were studied for methicillin resistance is 1993 and 1996. An increase in methicillin resistance among these isolates was observed in 1996. In 1993, 32.6 per cent isolates of S. aureus were methicillin resistant, this increased to 45.7 per cent in 1996. Methicillin resistance in CONS were 1.6 and 14.6 per cent respectively in 1993 and 1996. This increase in methicillin resistance may pose therapeutic problems and requires more effective drugs based on susceptibility testing of such isolates.

Analysis of sickle cell gene using polymerase chain reaction & restriction enzyme Bsu 361.

A 772bp DNA fragment from human beta-globin gene has been amplified by polymerase chain reaction (PCR) and subjected to restriction enzyme analysis using Bsu 361, an isoschizomer of restriction enzyme Mst II. This protocol has been designed basically to enhance the analytical facility for the detection of sickle cell mutation. A 430bp DNA fragment was found to be associated with the mutant locus, whereas 228bp and 202bp DNA fragments were generated from the normal locus. This difference of about 202bp in the resulting fragments from the mutant and normal loci has improved discriminatory power in the genotype analysis of the sickle cell mutation.

Measurement of dehabilitation in patients of leprosy--a scale.

Leprosy interferes with the psychological and social life of the patients leading to their 'dehabilitation'. Therefore, it is necessary to assess the extent and direction of dehabilitation in order to make the treatment plan comprehensive and effective. The objective of this work was to: (a) construct a scale for measuring dehabilitation and, (b) to standardize it. The methodology included preparation of 52 statements (in English) spread over four subareas of life, namely family relations, vocational condition, social interaction and self-esteem. It was administered to 122 randomly selected respondents. Scores were awarded by summing up the weights of each statement, a high score indicating low dehabilitation. Statistical tests were applied for standardizing the scale. To establish reliability, split-half reliability test and item discriminant analysis were used. Factor analysis was used to test the validity. The results show, that the split-half reliability coefficient ranged high (from 0.64 to 0.83) in all four subareas. The item discriminant analysis had a level of significance of 0.001 for 42 statements while the factor analysis explained variance covered over 70 percent. Hence the scale can be an useful intervention strategy for counselling, case work or rehabilitation.

Calcium activated neutral protease & calcium ATPase in glucose-6-phosphate dehydrogenase deficiency hemizygotes.

To study the mechanism of haemolysis in G6PD deficient erythrocytes, studies were undertaken in G6PD deficiency and in normal erythrocytes artificially loaded with calcium. Significantly increased concentrations of calcium, calcium activated neutral protease (CANP) and calcium ATPase were found in patients of G6PD deficiency. However, the membrane bound calcium, the total glycoprotein and sulphydryl groups of membrane were observed to be decreased. Similar results were also observed in the normal erythrocytes when loaded with calcium. These results point to the role of the proteolytic process in membrane modification, and altered membrane permeability during the haemolytic process. Our observations in G6PD deficiency and in in vitro point to the existence of a calcium dependent proteolytic preconditioning of erythrocyte accelerating the haemolysis.

Effect of Ep475 on changes induced by calcium activated neutral protease on erythrocyte membrane.

Increase in cytosolic calcium leads to activation of calcium activated neutral protease (CANP). CANP is known to cause many membrane abnormalities that aid erythrocyte destruction. An epoxy compound Ep475 causes the reversal of such changes induced by calcium and CANP. In absence of Ep475, CANP caused reduction in free sulfhydryl groups and glycoprotein content of the membrane to 61, and 50% respectively, compared to untreated membranes. Calcium ATPase and membrane associated CANP were increased 3 and 1.4 times respectively. Significant reversal of these changes by Ep475 suggests a possible role of this compound in reversing the calcium dependent alterations in RBC including the action of CANP.

Importance of monitoring calcium & calcium related properties in carrier detection for Duchenne muscular dystrophy.

Calcium and calcium dependent enzymes viz., calcium ATPase, protein kinase C and calcium activated neutral protease (milli CANP mCANP) were studied in the erythrocytes, platelets and lymphocytes of obligate carriers, in order to assess the usefulness of these indices for detection of carriers for Duchenne muscular dystrophy (DMD). With the exception of mCANP and lymphocyte calcium ATPase, other calcium dependent enzyme activities showed considerable overlap between carriers and control. Since the increase in the level of platelet mCANP was found in all affected boys (no false negatives) and obligate carriers, and patients with other myopathic conditions and some neurogenic causes did not show high platelet mCANP activity, this parameter could be considered as a good phenotypic index. Unlike SCK, the platelet mCANP of carriers did not overlap that of controls, hence tests are to be carried out to verify its usefulness as an index of carrier state in mutations other than DNA deletion since testing of non-deletion is both costly and has practical limitations.

Endogenous inhibitor of calcium activated neutral proteinase from human placenta: Purification and possible mechanism of proteinase regulation.

An endogenous inhibitor of calcium activated neutral proteinase has been purified from human placenta. The procedure included chromatography on DEAE cellulose, Ultrogel AcA 22 and milli calcium activated neutral proteinase-sepharose in succession. Endogenous calcium activated neutral proteinase inhibitor was a tetramer with identical subunits of molecular weight 68 kDa. It was specific for milli calcium activated neutral proteinase (Calpain II) which is inhibited by the formation of an inactive enzyme-inhibitor complex and not by sequestering Ca 2+ from the medium. Although micro calcium activated neutral proteinase (Calpain I) was not inhibited by endogenous calcium activated neutral proteinase inhibitor, it was protected from autolysis in the presence of the inhibitor. The placental endogenous calcium activated neutral proteinase inhibitor thus regulates Ca2+ activated proteolysis by ensuring micro calcium activated neutral proteinase activity, while inhibiting milli calcium activated neutral proteinase.

Human placental calcium activated neutral proteinase: Separation and functional characterization of subunits.

The subunits of human placental milli calcium activated neutral proteinase and micro calcium activated neutral proteinase have been separated by partial denaturation with urea followed by molecular sieving, with a recovery of 82–91% of activity. The separated subunits were homogeneous, as judged by sodium dodecyl sulphatepolyacrylamide gel electrophoresis. Their molecular sizes, catalytic activities and sulphydryl contents suggest that both the subunits of these two calcium activated neutral proteinases are distinct. The subunits were highly specific and could not be interchanged. Both the subunits of micro calcium activated neutral proteinase were catalytically active whereas only the 80 k subunit of milli calcium activated neutral proteinase was active. 30 k subunit of milli calcium activated neutral proteinase has a regulatory role since maximum activity of the 80 k subunit was elicited only in its presence. Activity of the reassociated subunits indicated that interaction is essential for the expression of optimum activity. Interaction of subunits rendered the enzymes less susceptible to inhibition by endogenous calcium activated neutral proteinase inhibitor.

Calcium activated neutral protease in blood cells from patients & carriers of Duchenne muscular dystrophy.

As blood cells such as platelets, lymphocytes and erythrocytes from patients with Duchenne muscular dystrophy show evidence of membrane alterations and elevation of intra-cellular calcium, one of the calcium related changes i.e., the activity of calcium activated neutral protease (CANP) was monitored and found to be elevated in erythrocytes, lymphocytes and platelets. As similar changes were observed in platelets of carriers of this disease, CANP in platelets may serve as a useful index for carrier detection.