Hibiscus sabdariffa Extract Inhibits Adhesion, Biofilm Initiation and Formation in Candida albicans.

Microbial biofilms act as reservoirs for pathogenic sessile microbes which reside inside the three dimensional matrix of the biofilm, and are thus protected against anti-microbial drugs. Most of the anti-microbial drugs fail to completely abolish the biofilm associated infections. In the present study, we provide evidence of Hibiscus sabdariffa (Hs) extract having possible anti-microbial activity, with emphasis on Candida albicans biofilm. The Hs extract was shown to be effective against C. albicans pre-formed biofilm at 3.125 mg/ml and was able to inhibit the hyphae initiation and adherence of cells. Furthermore, Hs extract was able to reduce the C. albicans load in C. elegans by effectively killing the Candida cells thereby reducing the viable colony count and effectively increasing the lifespan of worms. The percentage of viable hatched progeny of worms exposed to Hs extract (both at conc. 1.5 mg/ml and 6.25 mg/ml), was also comparable to that of the control untreated eggs. The Hs extract was also found to be significantly effective against fluconazole resistant C. albicans isolated from patients. Thus, we, for the first time, propose Hs extract as a prospective drug candidate and substitute for eradicating pre-formed biofilm and inhibiting the growth of C. albicans.

Issues in antifungal stewardship: an opportunity that should not be lost.

Many countries have observed an increase in the incidence of invasive fungal infections (IFIs) over the past two decades with emergence of new risk factors and isolation of new fungal pathogens. Early diagnosis and appropriate antifungal treatment remain the cornerstones of successful outcomes. However, due to non-specific clinical presentations and limited availability of rapid diagnostic tests, in more than half of cases antifungal treatment is inappropriate. As a result, the emergence of antifungal resistance both in yeasts and mycelial fungi is becoming increasingly common. The Delhi Chapter of the Indian Association of Medical Microbiologists (IAMM-DC) organized a 1 day workshop in collaboration with BSAC on 10 December 2015 in New Delhi to design a road map towards the development of a robust antifungal stewardship programme in the context of conditions in India. The workshop aimed at developing a road map for optimizing better outcomes in patients with IFIs while minimizing unintended consequences of antifungal use, ultimately leading to reduced healthcare costs and prevention development of resistance to antifungals. The workshop was a conclave of all stakeholders, eminent experts from India and the UK, including clinical microbiologists, critical care specialists and infectious disease physicians. Various issues in managing IFIs were discussed, including epidemiology, diagnostic and therapeutic algorithms in different healthcare settings. At the end of the deliberations, a consensus opinion and key messages were formulated, outlining a step-by-step approach to tackling the growing incidence of IFIs and antifungal resistance, particularly in the Indian scenario.

Multilocus sequence typing of Candida africana from patients with vulvovaginal candidiasis in New Delhi, India.

We investigated the prevalence of vulvovaginal candidiasis due to C. africana in an STD clinic in India and analysed the genetic relatedness of these C. africana isolates with those outside India. A total of 283 germ-tube-positive yeasts were identified by VITEK2. Molecular characterisation of all isolates was carried out by hwp1-gene-specific PCR. Of 283 germ-tube-positive yeast isolates, four were identified as C. africana using hwp1-gene-specific PCR. All hwp1 PCR positive C. africana were subjected to antifungal susceptibility testing, ITS and D1/D2 region sequencing and were typed by using MLST approach. Similar to C. africana isolates from the United Kingdom and unlike those from Africa, the Indian C. africana grew at 42°C. Sequencing of eight gene fragments in MLST identified all four strains to have different genotypes not reported previously. Furthermore, though the Indian C. africana isolates were susceptible to most of the 14 tested antifungal drugs, differences in susceptibility were observed among the four strains. Our results indicate genetic and phenotypic heterogeneity among C. africana from different geographical regions. Due to lack of data on epidemiology and genetic variability of this under-reported yeast, more studies using molecular methods are warranted.

Risk factors associated with increasing prevalence of gonorrhea and the antimicrobial susceptibility profiles of Neisseria gonorrhoeae among adolescents: A decade-long, hospital-based study from India.

Background: Gonorrhea is a significant cause of morbidity among sexually active population. Young adults and adolescents have a high risk of contracting sexually transmitted infections (STIs) due to behavioral, biological, and cultural factors fuelling the epidemic among them. The Centers for Disease Control recommends annual STI screening for sexually active individuals under twenty-five and those at higher risk. The present study aims at determining the risk factors linked to the rising prevalence of gonorrhea among Indian adolescents and antimicrobial susceptibility profiles of Neisseria gonorrhoeae. Materials and Methods: Clinical samples from adolescents attending the STI clinics were collected over 10 years. The isolates were confirmed as N. gonorrhoeae and antimicrobial susceptibility tests were performed to various drugs using the minimum inhibitory concentration strip method. Results: A total of 7308 genital discharge specimens were collected from STI clinic attendees, of which 417 samples were positive for gonorrhea (25 among male adolescents). Seventy six percent of positive adolescents had multiple sex partners, with only 4% using condoms. Nearly 20% practiced exchange of drugs for sex. Antimicrobial susceptibility rates were 96% sensitive for azithromycin, cefixime, and ceftriaxone. Gentamicin and spectinomycin reported 100% sensitivity rates. High resistance rates were reported to penicillin, ciprofloxacin, and tetracycline at 80%, 88%, and 68%, respectively. Conclusion: Regular screening for STIs helps understand the trends and transmission of gonorrhea, which helps initiate appropriate control measures. The resistance to recommended treatment regimens such as azithromycin and cefixime seems to be escalating gradually, probably due to irrational use of antibiotics for non-STI cases and empirical treatment, which needs close monitoring.

Genital Chlamydia trachomatis: an update.

Chlamydia trachomatis is the most common cause of curable bacterial sexually transmitted infection (STI) worldwide. It manifests primarily as urethritis in males and endocervicitis in females. Untreated chlamydial infection in man can cause epididymitis and proctitis. Though most women with Chlamydia infection are asymptomatic or have minimal symptoms, some develop salpingitis, endometritis, pelvic inflammatory disease (PID), ectopic pregnancy and tubal factor infertility. It is associated with an increased risk for the transmission or acquisition of HIV and is also attributed to be a risk factor for the development of cervical carcinoma. Early diagnosis and treatment of infected individuals is required to prevent the spread of the disease and severe sequelae. Traditionally, tissue culture was considered the gold standard for the diagnosis. However, with the availability of newer diagnostic techniques particularly molecular methods which are not only highly sensitive and specific but are cost-effective also, the diagnosis has became fast and easy. The purpose of this review is to study the various aspects of genital C. trachomatis infection. Also the advances related to the clinical picture, various diagnostic modalities, prevention, treatment, drug resistance and control measures will be dealt with.

Cinnamic aldehydes affect hydrolytic enzyme secretion and morphogenesis in oral Candida isolates.

Effect of cinnamaldehyde (CD), 4-hydroxy-3-methoxy cinnamaldehyde (HMCD) and 3,5-dimethoxy-4-hydroxy cinnamaldehyde (HDMCD) on growth and virulence factors of standard (Candida albicans 90028) and 26 oral isolates of C. albicans has been investigated. Growth was significantly inhibited by all three compounds in both solid and liquid medium, no systematic difference was observed between various isolates. MIC90 ranged from 125 to 450 µg/ml for CD, 100-250 µg/ml for HMCD and 62.5-125 µg/ml for HDMCD. All oral isolates were found to be proteinase and phospholipase secretors, both proteinase and phospholipase secretion was significantly inhibited by all the three tested molecules. No systematic difference in secretion or its inhibition was observed between standard and oral isolates as also between various isolates. Average drop in proteinase and phospholipase secretion caused by ½ MIC of CD was 33% and 28%, HMCD; 46% and 44%, HDMCD; 59% and 54%. The standard strain and all the 26 oral isolates displayed morphogenesis under triggering experimental conditions; no difference was seen between standard and various isolates. In the absence of test compounds hyphae development at 300 min was 83% for standard strain whereas average hyphae development for oral isolates was 85%. Average hyphal transition was suppressed by all tested compounds. At ½ MIC concentration at 300 min average hyphal transition of standard and oral isolates was CD; 49% and 57%, HMCD; 45% and 38%, HDMCD; 5% and 5%. Average haemolytic activity of the three tested compounds varied from 10 to 15% at their highest MIC compared to 20% shown by fluconazole at typical MIC of 30 µg/ml.

Proton pumping ATPase mediated fungicidal activity of two essential oil components.

This work evaluates the antifungal activity of two essential oil components against 28 clinical isolates (17 sensitive, 11 resistant) and 3 standard laboratory strains of Candida. Growth of the organisms was significantly effected in both solid and liquid media at different test compound concentrations. The minimum inhibitory concentrations (MICs) of Isoeugenol (compound 1) against 31 strains of Candida ranged 100-250 µg/ml and those of o -methoxy cinnamaldehyde (compound 2) ranged 200-500 µg/ml, respectively. Insight studies to mechanism suggested that these compounds exert antifungal activity by targeting H(+)-ATPase located in the membranes of pathogenic Candida species. At their respective MIC(90) average inhibition of H(+)-efflux for standard, clinical and resistant isolates caused by compound 1 and compound 2 was 70%, 74%, 82% and 42%, 42% and 43%. Respective inhibition of H(+)-efflux by fluconazole (5 µg/ml) was 94%, 92% and 10%. Inhibition of H(+)-ATPase leads to intracellular acidification and cell death. SEM analysis of Candida cells showed cell membrane breakage and alterations in morphology. Haemolytic activity on human erythrocytes was studied to exclude the possibility of further associated cytotoxicity.

Interesting anticandidal effects of anisic aldehydes on growth and proton-pumping-ATPase-targeted activity.

Attention has been drawn to evaluate the antifungal activity of p-anisaldehyde (1), o-anisaldehyde (2) and m-anisaldehyde (3). To put forward this approach, antifungal activity has been assessed in thirty six fluconazole-sensitive and eleven fluconazole-resistant Candida isolates. Growth and sensitivity of the organisms were significantly effected by test compounds at different concentrations. The rapid irreversible action of compound-1, compound-2 and compound-3 on fungal cells suggested a membrane-located target for their action. We investigated their effect on H(+) ATPase mediated H(+)-pumping by various Candida species. All the compounds inhibit H(+)- ATPase activity at their respective MIC(90) values. Inhibition of H(+) ATPase leads to intracellular acidification and cell death. Scanning electron microscopy analysis revealed deep wrinkles, deformity and flowed content. Furthermore, it was also observed that position of methoxy group attached to the benzene ring decides antifungal activity of the compound. The present study indicates that compound-1, compound-2 and compound-3 have significant antifungal activity against Candida, including azole-resistant strains, advocating further investigation for clinical applications in the treatment of fungal infections.

Needle stick injuries among health care workers in a tertiary care hospital of India.

BACKGROUND & OBJECTIVES: Percutaneous injuries caused by needlesticks, pose a significant risk of occupational transmission of bloodborne pathogens. Their incidence is considerably higher than current estimates, and hence a low injury rate should not be interpreted as a non existent problem. The present study was carried out to determine the occurrence of NSI among various categories of health care workers (HCWs), and the causal factors, the circumstances under which these occur and to, explore the possibilities of measures to prevent these through improvements in knowledge, attitude and practice. METHODS: The study group consisted of 428 HCWs of various categories of a tertiary care hospital in New Delhi, and was carried out with the help of an anonymous, self-reporting questionnaire structured specifically to identify predictive factors associated with NSIs. RESULTS: The commonest clinical activity to cause the NSI was blood withdrawal (55%), followed by suturing (20.3%) and vaccination (11.7%). The practice of recapping needles after use was still prevalent among HCWs (66.3%). Some HCWs also revealed that they bent the needles before discarding (11.4%). It was alarming to note that only 40 per cent of the HCWs knew about the availability of PEP services in the hospital and 75 per cent of exposed nursing students did not seek PEP. INTERPRETATION & CONCLUSIONS: The present study showed a high occurrence of NSI in HCWs with a high rate of ignorance and apathy. These issues need to be addressed, through appropriate education and other interventional strategies by the hospital infection control committee.

Profile of sexually transmitted infections among attendees of special clinics (Suraksha) at an apex regional sexually transmitted disease centre in North India ­ a five-year study

Introduction: Sexually transmitted infections (STIs) present significant global and national health challenges, particularly in India. Objective: To estimate the prevalence and characteristics of STIs among attendees at the Suraksha Clinic in the Apex Regional STD Centre, Safdarjung Hospital. Methods:Retrospective data from January 2018 to December 2022 were statistically analyzed using Excel and SPSS. The study included the examination for diagnosis of various STIs, such as syphilis, human immunodeficiency virus (HIV), gonorrhoea, chlamydiasis, trichomoniasis, candidiasis, bacterial vaginosis, chancroid, and genital herpes. Gender distribution and syndromic diagnoses, including vaginal/cervical discharge and genital ulcers, were also considered. Referrals to Integrated Counseling and Testing Centres for HIV testing were analyzed. Results: The outcomes reveal a substantial burden of STIs, with 3.06% showing reactivity to syphilis, 1.74% testing positive for HIV, 3.36% for gonorrhoea, 11.78% for chlamydiasis, 1.05% for trichomoniasis, 26.24% for candidiasis, 9.97% for bacterial vaginosis, 7.80% for chancroid, 11.64% for herpes genitalis, and 4.01% for other non-STIs. Attendees' interactions included 34.36% of referrals to Integrated Counseling and Testing Centres for HIV testing. The gender distribution showed 58.92% male and 40.94% female attendees. Conclusion: Syndromic diagnoses, including vaginal/cervical discharge (21.22%) and genital warts (8.00%), highlight prevalent conditions, necessitating routine screening, early detection, and targeted interventions for effective disease control and prevention. These findings underscore the significance of integrated screening, patient education, and proactive strategies to safeguard public health in the face of rising STI rates. (AU)

Introdução: As infecções sexualmente transmissíveis (IST) apresentam desafios de saúde globais e nacionais significativos, particularmente na Índia. Objetivo: Estimar a prevalência e as características das IST entre os pacientes atendidos na Clínica Suraksha do Centro Regional de doenças sexualmente transmissíveis (DST) Apex, Hospital Safdarjung. Métodos: Dados retrospectivos de janeiro de 2018 a dezembro de 2022 foram analisados estatisticamente utilizando Excel e Statistical Package for the Social Sciences (SPSS). O estudo inclui o exame para diagnóstico de diversas IST, como sífilis, HIV, gonorreia, clamídia, tricomoníase, candidíase, vaginose bacteriana, cancroide e herpes genital. A distribuição por gênero e os diagnósticos sindrômicos, incluindo corrimento vaginal/cervical e úlceras genitais, também foram considerados. Foram analisados encaminhamentos para Centros Integrados de Aconselhamento e Testagem para testagem de HIV. Resultados: Os resultados revelam uma carga substancial de IST, com 3,06% apresentando reatividade à sífilis, 1,74% testando positivo para HIV, 3,36% para gonorreia, 11,78% para clamídia, 1,05% para tricomoníase, 26,24% para candidíase, 9,97% para vaginose bacteriana, 7,80% para cancroide, 11,64% para herpes genital e 4,01% para outras infecções não IST. As interações dos participantes incluíram 34,36% de encaminhamentos para Centros Integrados de Aconselhamento e Testagem para testes de HIV. A distribuição por gênero mostrou 58,92% de participantes do sexo masculino e 40,94% do sexo feminino. Conclusão: Os diagnósticos sindrômicos, incluindo corrimento vaginal/cervical (21,22%) e verrugas genitais (8,00%), destacam condições prevalentes, necessitando de exames de rotina, detecção precoce e intervenções direcionadas para controle e prevenção eficazes de doenças. Estas conclusões sublinham a importância do rastreio integrado, da educação dos pacientes e de estratégias proativas para salvaguardar a saúde pública diante do aumento das taxas de IST. (AU)

A novel polyherbal microbicide with inhibitory effect on bacterial, fungal and viral genital pathogens.

A polyherbal cream (Basant) has been formulated using diferuloylmethane (curcumin), purified extracts of Emblica officinalis (Amla), purified saponins from Sapindus mukorossi, Aloe vera and rose water along with pharmacopoeially approved excipients and preservatives. Basant inhibits the growth of WHO strains and clinical isolates of Neisseria gonorrhoeae, including those resistant to penicillin, tetracycline, nalidixic acid and ciprofloxacin. It has pronounced inhibitory action against Candida glabrata, Candida albicans and Candida tropicalis isolated from women with vulvovaginal candidiasis, including three isolates resistant to azole drugs and amphotericin B. Basant displayed a high virucidal action against human immunodeficiency virus HIV-1NL4.3 in CEM-GFP reporter T and P4 (Hela-CD4-LTR-betaGal) cell lines with a 50% effective concentration (EC50) of 1:20000 dilution and nearly complete (98-99%) inhibition at 1:1000 dilution. It also prevented the entry of HIV-1(IIIB) virus into P4-CCR5 cells (EC50 approximately 1:2492). Two ingredients, Aloe and Amla, inhibited the transduction of human papillomavirus type 16 (HPV-16) pseudovirus in HeLa cells at concentrations far below those that are cytotoxic and those used in the formulation. Basant was found to be totally safe according to pre-clinical toxicology carried out on rabbit vagina after application for 7 consecutive days or twice daily for 3 weeks. Basant has the potential of regressing vulvovaginal candidiasis and preventing N. gonorrhoeae, HIV and HPV infections.

CD4/CD8 lymphocyte counts in healthy, HIV-positive individuals & AIDS patients.

BACKGROUND & OBJECTIVES: The enumeration of CD4 and CD8 positive cells, surrogate markers for HIV disease progression, is helpful in management and follow up of immunocompromised HIV-positive patients. In assessing the degree of immune deficiency in HIV-positive patients of a particular region, knowledge of reference range of T-cell subset counts of healthy individuals of that particular region is essential. The present cross-sectional study was undertaken to determine the reference range of T-cell subsets in healthy north Indians and to compare the values with those in HIV-positives. METHODS: Blood samples from 125 HIV seronegative healthy volunteers comprising group I (88 males, 37 females) and 452 HIV- positive patients, divided into group II of asymptomatic (n=137; 93 males, 44 females) and group III of AIDS patients (n=315; 253 males, 62 females) in the age group of 17-60 yr, were analysed for enumeration of CD4+, CD8+ cells/microl by flow cytometry. RESULTS: In group I, the CD4 and CD8 levels were 687 +/- 219 and 611 +/- 288 cells/microl in males and 740 +/- 255 and 546 +/- 246 cells/microl in females. Overall, a significant depressed level of CD4 (525 +/- 207 cells/microl) and elevated level of CD8 (1174 +/- 484 cells/microl) in group II and (170 +/- 115 and 1051 +/- 586 cells/microl) respectively in group III were observed. Group II patients had highest level of CD8 cells. No asymptomatic women had CD4 count of <200 cells/microl. INTERPRETATION & CONCLUSION: Our findings on T-cell subset reference ranges of normal healthy north Indians validate the utility of determination of CD4 cell count as a useful predictor of AIDS in Indian conditions and confirm that a significant per cent of AIDS patients had CD4 cell count below 200/microl.

A situational analysis of sexual and reproductive health issues in physically challenged people, attending a tertiary care hospital in New Delhi.

BACKGROUND AND OBJECTIVES: Physically challenged people constitute the most stigmatized sections of society, and are excluded from outreach programs, besides being considered sexually inactive. They have unaddressed sexual and reproductive healthcare (SRH) issues, predisposing them to sexual abuse and sexually transmitted infections (STIs). The huge paucity of data in this field prompted us to undertake this study. MATERIALS AND METHODS: A cross-sectional survey was conducted on 100 people with more than 40% of permanent disability, attending various out/inpatient facilities of a tertiary care hospital in New Delhi, India. A structured, pretested questionnaire was used to assess SRH issues. Samples were collected from consenting individuals for diagnosis of various STIs, wherever relevant. Statistical analysis was done using Pearson's Chi-square test, considering significant at P <0.05. RESULTS: Most people were in the age group of 15-30 years. Limbs were most commonly affected, and the use of assistive devices was statistically related to income levels (P = 0.045), 43% was married and 41% had children. Contraceptive usage was 33%, with a significant association (P = 0.03) with education levels. Issues related to sexual health included conditions ranging from nerve sensation loss in genitalia to fertility and gynecological issues, only 10% had received sexual counseling during rehabilitation. There were several misconceptions prevalent regarding HIV and STIs; 35% of the samples tested positive for chlamydia IgG. INTERPRETATION AND CONCLUSIONS: This is a pioneer study on a grossly neglected issue in India. There is a dire need to overcome hurdles and address the SRH issues of physically challenged people to achieve the universal WHO goal of "Health for All."

Molecular methods in the laboratory diagnosis of sexually transmitted infections.

Sexually transmitted infections (STIs) are a public health problem, and their prevalence is rising even in developed nations, in the era of HIV/AIDS. While the consequences of STIs can be serious, the good news is that many of these complications are preventable if appropriate screening is done in high-risk individuals, when infection is strongly suspected. The diagnostic tests for STIs serve many purposes. Apart from aiding in the diagnosis of typical cases, they help diagnose atypical cases, asymptomatic infections and also multiple infections. But, the test methods used must fulfill the criteria of accuracy, affordability, accessibility, efficiency, sensitivity, specificity and ease of handling. The results must be rapid, cost-effective and reliable. Most importantly, they have to be less dependent on collection techniques. The existing diagnostic methods for STIs are fraught with several challenges, including delay in results, lack of sensitivity and specificity. With the rise of the machines in diagnostic microbiology, molecular methods offer increased sensitivity, specificity and speed. They are especially useful for microorganisms that cannot be, or are difficult to cultivate. With the newer diagnostic technologies, we are on the verge of a major change in the approach to STI control. When diagnostic methods are faster and results more accurate, they are bound to improve patient care. As automation and standardization increase and human error decreases, more laboratories will adopt molecular testing methods. An overview of these methods is given here, including a note on the point-of-care tests and their usefulness in the era of rapid diagnostic tests.

Diversity and antifungal resistance patterns of prevalent opportunistic pathogenic yeasts colonizing the oral cavities of asymptomatic human immunodeficiency virus-infected individuals, and their relation to CD4(+) counts.

BACKGROUND: Yeasts are important opportunistic pathogens, in individuals infected with human immunodeficiency virus (HIV). Yeast species inhabiting the oral mucosa of HIV-infected persons can act as source of oral lesions, especially as the individual progresses towards immunocompromised state. Present study was conducted to evaluate the diversity of yeasts in oral cavities of asymptomatic HIV-infected persons and their association with CD4(+) cell counts. MATERIALS AND METHODS: 100 HIV seropositive subjects and 100 healthy controls were screened for oral yeast carriage using standard procedures. RESULTS: Of the 100 HIV-seropositive persons screened, 48 were colonized by different yeasts, either alone or in association with another species. Candida albicans was the most common species (56.90%) while non C. albicans Candida (NCAC) accounted for 39.65%. Among NCAC, Candida tropicalis and Candida krusei were most common. One isolate each of rare opportunistic pathogenic yeasts, Geotrichum candidum and Saccharomyces cereviseae, was recovered. The control group had an oral candidal carriage rate of 23%; C. albicans was the predominant species, followed by Candida glabrata, C. tropicalis and Candida parapsilosis. Antifungal susceptibility testing revealed no resistance in C. albicans, to the commonly used antifungal agents, whereas resistance or dose dependent susceptibility to fluconazole was observed in some of the NCAC species. CONCLUSION: Oral carriage of opportunistic pathogenic yeasts was greater in HIV-seropositive persons heading towards immunocompromised state, as evidenced by their CD4(+) cell count. The predominant yeast isolated in this study (C. albicans), was found to be susceptible to commonly used antifungals.

Evaluation of three methods to determine the antimicrobial susceptibility of Mycobacterium tuberculosis.

BACKGROUND & OBJECTIVES: Tuberculosis continues to be a major public health problem in India, especially with the emergence of drug resistance. A study was carried out to establish a rapid and accurate method of susceptibility testing for Mycobacterium tuberculosis using three methods viz., proportion method by agar dilution on Middlebrook 7H11 agar, proportion method using the conventional Lowenstein-Jensen (L-J) medium and E test strip method. METHODS: A total of seventy five clinical isolates from pulmonary and extrapulmonary sites were characterised and speciated by biochemical tests, growth and other standard parameters, and eight random isolates, also by polymerase chain reaction (PCR). Antimicrobial susceptibility of M.tuberculosis was performed by proportion method on L-J medium and Middlebrook 7H11 agar medium for isoniazide (INH), rifampicin (RIF), ethambutol (EMB), streptomycin (STM) and ciprofloxacin (CIP) using recommended critical concentrations. The two methods were compared with the E test method. RESULTS: The 75 M.tuberculosis strains were isolated from sputum (47), pus (23), aspirate fluid (2), skin tissue (2) and gastric aspirate (1). Of these 49 (65.3%) isolates were sensitive and one (1.3%) was resistant to all the five drugs tested and by all the three methods. Eleven (14.7%) isolates were resistant to INH alone by the three methods. The E test method detected one isolate resistant to INH and 2 to RIF which were missed by the other two methods. The results obtained by all the three methods compared well. INTERPRETATION & CONCLUSION: The three methods viz., proportion methods with L-J, Middlebrook 7H11 agar and the E test concurred fully in 57 isolates (76%). Association between L-J and Middlebrook 7H11 agar methods was 59 per cent. E test and the L-J methods did not differ significantly for all the drugs. The finding show that the E test method is superior to the other two methods in terms of simplicity of performance and the rapidity of results. Another advantage is that the MIC values can also be obtained simultaneously by this method.

Sexual resilience within intimate relations among unmarried adolescent girls seeking abortion in an abortion clinic of Delhi, India.

BACKGROUND AND OBJECTIVES: Sexually experienced, unmarried adolescent women, usually commence sex, with marriage in mind. Initially, they resist sex but are unsuccessful due to lack of resilience to end an oppressive relationship, or convince their partners to postpone sex or use protection. To retain partners, they accept unprotected sex and suffer its consequences such as unwanted pregnancies, sexually transmitted diseases, HIV/AIDS and cervical cancer. Considerable numbers of adolescents face this emotional stress and suffer the consequences, while some manage to endure and emerge from the ordeal. This study attempts to determine this resilience and extrapolate it to others who might not be so successful without an intervention. METHODOLOGY: The study compares sexual resilience in 100 unmarried adolescent abortion-seekers, in terms of time taken from meeting the partner, to sexual debut and correlates this with background factors such as age, education, family income, self-esteem, sexual knowledge, attitude toward pre-marital sex (PMS) and partner pressure. RESULTS: Mean debut age was 17.32 years. Partners took more initiative to form emotional (64%) and physical relationship (78%). Adolescent girls' initiative reduced markedly from emotional (22%) to physical relation (5%). Correlation of sexual resilience with age, family income education and knowledge was not significant. It was significantly correlated with attitude toward PMS, self-esteem and partner pressure. INTERPRETATION AND CONCLUSION: It is desirable to improve self-esteem and attitude of young women to build negotiation skills in intimate relations. Counselors have to reorient the perspectives to improve attitude toward abstinence and increase self-esteem to resist pressure from partners.

Influences of cinnamic aldehydes on H⁺ extrusion activity and ultrastructure of Candida.

The antifungal effects of cinnamaldehyde, 4-hydroxy-3-methoxycinnamaldehyde (coniferyl aldehyde) and 3,5-dimethoxy-4-hydroxycinnamaldehyde (sinapaldehyde) were investigated against 65 strains of Candida (six standard, 39 fluconazole-sensitive and 20 fluconazole-resistant). MICs of cinnamaldehyde, coniferyl aldehyde and sinapaldehyde ranged from 100 to 500 µg ml(-1), 100 to 300 µg ml(-1) and 100 to 200 µg ml(-1), respectively. All tested isolates showed a marked sensitivity towards these aldehydes in spot and time-kill assays. Sinapaldehyde was found to be the most effective, followed by coniferyl aldehyde and cinnamaldehyde. At their respective MIC(90) values, the three compounds caused mean inhibition levels of glucose-stimulated H(+)-efflux of 36, 34 and 41 % (cinnamaldehyde), 41, 42 and 47 % (coniferyl aldehyde) and 43, 45 and 51 % (sinapaldehyde) for standard-sensitive, clinical-sensitive and clinical-resistant isolates, respectively. Inhibition levels of H(+)-efflux caused by plasma membrane ATPase inhibitors N,N'-dicyclohexylcarbodiimide (100 µM) and diethylstilbestrol (10 µM) were 34, 45 and 44 %, and 57, 39 and 35 %, for standard-sensitive, clinical-sensitive and clinical-resistant isolates, respectively. Intracellular pH (pHi) was found to decrease by 0.34, 0.42 and 0.50 units following incubation with three tested aldehydes from the control pHi of 6.70. Scanning electron microscopy and transmission electron microscopy analysis was performed on a representative strain, C. albicans 10261, showing alterations in morphology, cell wall, plasma membrane damage and lysis. Haemolytic activity of the three compounds varied from 10 to 15 % at their highest MIC compared to an activity level of 20 % shown by fluconazole at 30 µg ml(-1). In conclusion, this study shows significant activity of cinnamic aldehydes against Candida, including azole-resistant strains, suggesting that these molecules can be developed as antifungals.

Genital Ulcer Disease: How Worrisome Is It Today? A Status Report from New Delhi, India.

Background and Objectives. Genital ulcer diseases represent a diagnostic dilemma, especially in India, where few STI clinics have access to reliable laboratory facility. The changing STI trends require that a correct diagnosis be made in order to institute appropriate treatment and formulate control policies. The objective of this study was to determine recent trends in aetiology of genital ulcers, by using accurate diagnostic tools. Methods. Specimens from 90 ulcer patients were processed for dark field microscopy, stained smears, culture for H. ducreyi, and real-time PCR. Blood samples were collected for serological tests. Results. Prevalence of GUD was 7.45 with mean age at initial sexual experience as 19.2 years. Use of condom with regular and nonregular partners was 19.5% and 42.1%, respectively. Sexual orientation was heterosexual (92.2%) or homosexual (2.2%). There were 8 cases positive for HIV (8.9%). Herpes simplex virus ulcers were the commonest, followed by syphilis and chancroid. There were no cases of donovanosis and LGV. Conclusions. A valuable contribution of this study was in validating clinical and syndromic diagnoses of genital ulcers with an accurate aetiological diagnosis. Such reliable data will aid treatment and better define control measures of common agents and help eliminate diseases amenable to elimination, like donovanosis.