Inhibition of growth and virulence of Vibrio cholerae by carvacrol, an essential oil component of Origanum spp.

AIMS: In the age where bacterial resistance to conventional antibiotics is increasing at an alarming rate, the use of the traditional plant, herb extracts or other bioactive constituents is gradually becoming popular as an anti-virulence agent to treat pathogenic diseases. Carvacrol, a major essential oil fraction of Oregano, possesses a wide range of bioactivities. Therefore, we aimed to study the effect of sub-inhibitory concentrations of carvacrol on major virulence traits of Vibrio cholerae. METHODS AND RESULTS: We have used in vitro as well as ex vivo models to access the anti-pathogenic role of carvacrol. We found that the sub-inhibitory concentration of carvacrol significantly repressed bacterial mucin penetrating ability. Carvacrol also reduced the adherence and fluid accumulation in the rabbit ileal loop model. Reduction in virulence is associated with the downregulated expression of tcpA, ctxB, hlyA and toxT. Furthermore, carvacrol inhibits flagellar synthesis by downregulating the expression of flrC and most of the class III genes. CONCLUSIONS: Carvacrol exhibited anti-virulence activity against V. cholerae, which involved many events including the inhibition of mucin penetration, adhesion, reduced expression of virulence-associated genes culminating in reduced fluid accumulation. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that carvacrol possesses inhibitory activity against V. cholerae pathogenesis and might be considered as a potential bio-active therapeutic alternative to combat cholera.

Rhinoscleroma Revisited

ABSTRACT Rhinoscleroma (more appropriately 'scleroma') is a chronic, disfiguring inflammatory lesion that is rarely encountered in the present-day otolaryngology practice. The diagnosis often becomes difficult especially when it recurs and presents late with complications. This report illustrates the clinical and histologic features of rhinoscleroma in a defaulter patient who represents advanced-stage disease with orbital and intracranial extensions. Special emphasis has been provided on the computed tomography impressions such that the evolution of scleroma with time can be studied.

NMR-based metabonomics for understanding the influence of dormant female genital tuberculosis on metabolism of the human endometrium.

STUDY QUESTION: Does investigation of metabolic perturbations in endometrial tissue of women with dormant genital tuberculosis (GTB) during the window of implantation (WOI) assist in improving the understanding of endometrial receptivity? SUMMARY ANSWER: In dormant GTB cases significant alterations in endometrial tissue metabolites occur, largely related to energy metabolism and amino acid biosynthesis in dormant GTB cases. WHAT IS KNOWN ALREADY: As an intracellular pathogen, Mycobacterium tuberculosis strongly influences the metabolism of host cells causing metabolic dysregulation. It is also accepted that dormant GTB impairs the receptive status of the endometrium. Global metabolic profiling is useful for an understanding of disease progression and distinguishing between diseased and non-diseased groups. STUDY DESIGN, SIZE, DURATION: Endometrial tissue samples were collected from patients reporting at the tertiary infertility care center during the period September 2011-March 2013. Women having tested positive for GTB were considered as the study group (n = 24). Normal healthy women undergoing sterilization (n = 26) and unexplained infertile women with repeated IVF failure (n = 21) volunteered to participate as controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: Endometrial tissue samples were collected 6-10 days after confirmation of ovulation. PCR and BACTEC-460 culture were used for diagnosing GTB. Proton nuclear magnetic resonance (1H NMR) spectra of tissue were recorded using a 700 MHz Bruker Avance AV III spectrometer. Following phase and baseline correction of all NMR spectra by Bruker Topspin 2.1 software, spectral peak alignment of the data was performed. Multivariate analysis was applied to all spectra and individual metabolites identified and multiple correlation analysis was performed. MAIN RESULTS AND THE ROLE OF CHANCE: Leucine, isoleucine, acetate, lactate, glutamate, glutamine, methionine, lysine, creatine, glycogen, glycine, proline and choline were found to be significantly increased (P < 0.05) in endometrial tissue of women with dormant GTB compared with unexplained infertile women with repeated implantation failure. Valine, citrate, succinate and aspartate were also observed to be significantly up-regulated (P < 0.01). Furthermore, a significant decrease in glucose (P < 0.05), threonine (P < 0.05), tyrosine (P < 0.01) and phenylalanine (P < 0.0001) was observed in women with dormant GTB. Pearson's correlation analysis between the expression of various endometrial receptivity markers and metabolites showed a significant negative correlation (-0.236 to -0.545, P < 0.05). Also, the metabolites were positively correlated with endometrial receptivity markers (0.207 to 0.618, P < 0.05). LIMITATIONS, REASONS FOR CAUTION: It is often difficult to diagnose dormant GTB because it tends to exist without any clinical signs or symptoms. In addition, the diagnosis of GTB by culture remains a challenge due to low detection rates and its paucibacillary nature. Testing for prostate-specific antigen or the Y chromosome in order to account for the possible influences of recent exposure to semen on endometrial metabolism would be important. WIDER IMPLICATIONS OF THE FINDINGS: The metabolic changes associated with the dormant tubercle infection are of potential relevance to clinicians for the treatment of dormant GTB-related infertility. STUDY FUNDING/COMPETING INTERESTS: Government of India, Indian Council of Medical Research. There are no conflicts of interest.

Variable Presentations of Sinonasal Polypoid Masses: A Tertiary Institution Experience.

Background Lesions of the sinonasal area are varied, but they mostly present as polypoid masses which require meticulous work-up to reach at the most probable diagnosis. Objective Analysis of polypoid sinonasal masses in terms of etiology, clinical presentations, brief demographic profile, clinico-histologic correlate where possible, and follow-up results. Method In this descriptive, longitudinal study, 198 patients with polypoid sinonasal masses attending the otolaryngology clinic of a tertiary teaching institute were selected using proper selection criteria and analyzed through a pre-set proforma and algorithm for a diagnostic work-up (that included histopathology where necessary). Result Common presentations were nasal obstruction (~89%), discharge (~70%) and hyposmia (~22%). Though nearly 87% was clinically benign and 8% indeterminate, therapeutic and diagnostic interventions (including histopathology) showed 91% truly benign, of which polyposis formed the bulk. Sensitivity of clinical detection was 75% for benign lesions and 62% for malignancies. Diagnosis depended on histopathology in 52.52% cases, including the clinically malignant, the "grey zone", and more than 40% of the clinically benign lesions. There was male predilection (2.16 for benign lesions and 1.57 for malignant), rural preponderance, and above 60% of the patients were within 50-70 years. There was ~26% recurrence in the follow-up period of a minimum of one year, predominantly in polyposis (29.55%) and malignancies (~39%). Conclusion Presentations of polypoid sinonasal masses are variable, etiology of which is mostly benign. Proper clinico-histologic correlate is necessary for correct diagnosis. A low threshold of suspicion is required because of this variability, necessitating follow-up for further evaluation.

Detection, discrimination and discovery of a new Tobacco streak virus strain.

Soybean plants that exhibited symptoms of virus infection were sampled from different counties of Oklahoma. These plants were tested serologically for 15 viruses known to infect soybean plants. Fifty-seven samples that exhibited typical virus-like symptoms did not test positive for any of the 15 viruses used in a dot-immunobinding assay (DIBA). Four samples were pooled and used for next generation sequencing using the 454-Roche protocol. Sequence and phylogenetic analysis of the sequences obtained revealed infection with a distinct strain of Tobacco streak virus (TSV). TSV was one of the 15 viruses initially tested for using DIBA and had tested negative. TSV belongs to the genus Ilarvirus and has been reported as a causal agent of bud blight in soybean crops in Brazil and the United States. Out of 10 reported primer pairs for TSV reverse transcription-polymerase chain reaction (RT-PCR), only two had the potential, based on sequence similarity, to amplify part of the genome of the distinct strain of TSV found in Oklahoma and only one was actually able to amplify the region. In this study, a new primer pair, specific to all known TSV and capable of amplifying the Oklahoma strain (TSV-OK), was designed from a highly conserved region of coat protein (CP) sequences and end-point PCR and quantitative RT-PCR detection methods were developed and their sensitivity assayed. This is the first report of specific primers designed from this highly conserved region in the CP of TSV for detection of TSV. Twenty-three of the 57 DIBA soybean samples that initially tested negative were retested with the new specific end-point PCR method and found positive for TSV infection.

Primary soft tissue Ewing's sarcoma of the maxillary sinus in elderly patients: presentation, management and prognosis.

Nonosseous or soft tissue Ewing's sarcoma is a rare form of Ewing's sarcoma/primitive neuroectodermal tumour that seldom affects the head and neck region. Involvement of the nose and paranasal sinuses is extremely uncommon, with only eight of such patients being reported to date, mostly affecting adolescents and young adults. To our knowledge, this study is the first comprehensive report of primary soft tissue Ewing's sarcoma involving the paranasal sinuses in an elderly patient who successfully completed treatment. We herein discuss the pathogenesis, management and factors affecting the prognosis of this rare group of tumours involving the nose and paranasal sinuses, in relation to the available literature.

Radio-decontamination efficacy and safety studies on optimized decontamination lotion formulation.

Objective of the present study was to optimize decontamination lotion and to evaluate its relative decontamination efficacy using three radio-isotopes (Technetium-99m, Iodine-131 and Thallium-201) as contaminants with varying length of contaminant exposure (0-1h). Experiments were performed on Sprague Dawley rat's intact skin and human tissue equivalent models. Rat's hair was removed by using depilator after trimming with scissors. Relative decontamination efficacy of the optimized lotion was investigated and compared with water as control. Static counts were recorded before and after decontamination using single photon emission computed tomography (SPECT). Measured decontamination efficacy (DE) values were analyzed using one way ANOVA and Student's t-test (p value<0.05) and were found statistically significant. Decontamination efficacy of the lotion was observed to be 90 ± 5%, 80 ± 2% and 85 ± 2%, for the (131)I, (201)Tl and (99m)Tc radio-contaminants respectively on skin. Reduced contaminant removal was recorded for the skin which was cleaned by depilator (50-60%). Skin decontamination was found more efficacious for rat skin decontamination than the human tissue equivalent model. Decontamination efficacy of the lotion against (99m)Tc was recorded 70 ± 15% at 0-1h on the tissue equivalent model. In vitro chelation efficacy of the lotion was also established by using the instant thin layer chromatography-slica gel (ITLC-SG) and >95% of (99m)Tc was recorded. Neither erythema nor edema was scored in the primary skin irritancy test visually observed for two weeks.

Large, solitary angiokeratoma in the posterior third and base of the tongue: case report.

OBJECTIVE: We present an extremely rare case of isolated angiokeratoma of the tongue. METHOD: Case report and review of related literature. RESULTS: An 18-year-old, male adolescent presented with a fleshy, intermittently bleeding mass in the posterior third and base of the tongue. The lesion was initially suspected to be a lingual thyroid or haemangioma, but histopathological features were consistent with angiokeratoma. Magnetic resonance imaging revealed that the lesion extended up to the vallecula and involved the lamina propria and superficial tongue musculature. No similar lesions were found elsewhere in the body. No metabolic derangements were identified in the patient or his family. The 2.6 × 1.5 × 0.5 cm mass was excised under general anaesthesia. CONCLUSION: We present the 1st case of isolated lingual angiokeratoma in a male, the 4th such case overally, the largest ever documented. The lesion was situated in the posterior third and base of the tongue, a position not previously described.

Response to growth hormone therapy in Indian patients.

OBJECTIVE: To analyse response to growth hormone therapy on Indian patients with short stature. METHODS: Data were collected on 71 patients of short stature on GHT. All patients underwent clinical and hormonal evaluation. GHD was diagnosed in the presence of short stature (height SDS < 2) and peak GH levels < 10 ng/ml. Bone age was estimated using Tanner Whitehouse 3 method (TW3). RESULTS: Primary GHD (73%) was the commonest diagnosis among patients on GHT, followed by organic GHD (12.6%), genetic syndromes (8.4%) and systemic diseases (5.4%). Mean chronological age at presentation was 10.07+/-3.26 years (median-11 years, range 3-15 years), mean height age was 6.98+/-2.82 years (median 7.5 years, range 1-13 years) and mean bone age (available for 55 patients) was 7.19+/-3.1 years (median 8.2 years, range 1.3-13 years). Patients with systemic diseases (6.75+/-3.5 years) presented earlier, compared to patients with GHD (10.27+/-3.16 years) and genetic syndromes (10.18+/-3.20 years) (p=0.349). Most of the patients on GHT were in the age group 9-15 years (60.6%). Mean height gain with GHT was 8.7+/-2.7 cm (median 8.3 cm, range 3.0-13 cm) during 1st year then decreased to 6.9+/-2.4 cm (median 7.0 cm, range 3.0-12.5 cm) in the second year, and was maintained through the third year (mean 7.1+/-3.0 cm, median 7.0, range 3.0-13 cm). Among patients with GHD, those with primary deficiency had significantly better response to GHT in 1st year than secondary deficiency (9.0+/-2.65 vs 6.8+/-3.03 cm, p = 0.026). Response to GHT was negatively correlated with CA (r-0.27, p = 0.05), HA (r-0.47, p = 0.027) and BA (r-0.31, p=0.022) at presentation. Four patients (5.6%) developed hypothyroidism and one patient each developed disseminated tuberculosis and rickets. One patient of Turner's syndrome died of adrenal carcinoma. Short follow up and absence of measurement of IGF-1 and IGFBP3 were major limitations of this study. CONCLUSIONS: Response to GHT in Indian patients is comparable to western counterparts. Maximum height gain on GHT is during the first year than decreases in second year, but is maintained through third year. Patients with primary GHD had better response than secondary GHD. Response to GHT is negatively correlated with chronological, height and bone age at presentation.

Neonatal characteristics as risk factors for preschool asthma.

Childhood asthma usually begins early in life. Neonatal characteristics are reportedly predictive of symptom onset. This investigation utilized data from a provincial health organization to evaluate the effect of several birth characteristics on asthma incidence and hospitalization for asthma during age 0-4. Using logistic regression, the odds ratios (OR) for the following variables indicate a significant (p < 0.05) association with physician-diagnosed preschool asthma: male gender (OR = 1.72), birthweight < 1500 g (OR = 2.11), prematurity (OR = 1.34), respiratory distress syndrome (RDS) in the presence (OR = 2.95) or absence (OR = 1.61) of bronchopulmonary dysplasia (BPD), and transient tachypnea of the newborn (TTN; OR = 1.36). Male gender (OR = 1.91), birthweight < 1500 g (OR = 2.56), RDS with and without BPD (OR = 3.35 and 2.50, respectively), TTN (OR = 2.08), and severe birth asphyxia (OR = 1.94) showed an important association with hospitalization due to asthma. Neonatal characteristics are important determinants for the risk of preschool asthma, even after mutual adjustment.

Neoglycosylated liposomes as efficient ligands for the evaluation of specific sugar receptors on macrophages in health and in experimental leishmaniasis.

Receptors interacting with terminal sugars as ligands are involved in the binding of Leishmania donovani promastigotes to the macrophage surface and their subsequent internalization. Mannose and glucose are specifically involved in the binding process. Decreased binding occurs to macrophages already infected with L. donovani either in vivo or in vitro. When mannose- or glucose-bearing liposomes are used as ligands the binding shows similar trends and the percentage inhibition of binding with mannose-bearing liposomes increases when compared to that for the glucose-bearing ones. The decreased binding of the ligand seems to be due to a decrease in the number of receptors after infection. The affinity of the ligands for the binding sites either on the normal macrophages or on the infected macrophages apparently remains the same. The results based on the incorporation of [3H]phenyl alanine and supported by the binding of glycosylated liposomes to both infected and non-infected macrophages suggest that protein synthesis, in general, is suppressed in L. donovani-infected macrophages thus affecting also mannose/glucose receptor protein synthesis, resulting in fewer receptors on the macrophage surface.

An investigation of the increase in preschool-age asthma in Manitoba, Canada.

Asthma has long been a major cause of illness and disability among young Canadians. From 1970-71 to 1987-88, hospital admissions for asthma increased significantly among Canadian children under the age of fourteen. Many hypotheses may explain this increase in asthma prevalence. There could be a true increase in the number of people developing symptoms of the disease or increased asthma rates could be an artifact due to changes in detection, diagnosis, treatment, or coding. This study reviews hypotheses put forward to explain the increase in asthma prevalence, and tests some of them in Manitoba for children aged 0-4. Physician claims data and hospital separation data were merged to create unique person oriented medical records. These records were used to estimate the number of children seeking medical services for asthma during a five-year period (1984-85 to 1988-89) and the change in rates over this time period. From 1984-85 to 1988-89, both prevalence and incidence rates for children less than five years of age increased. Prevalence rates showed strong seasonal peaks in the spring and the fall. There is no indication that asthma increased in severity. The hospitalization rate (the number visiting a hospital for asthma divided by the total number seeking medical care for asthma), the average number of hospital admissions per year, and the average number of days spent in a hospital per year did not increase. Levels of ozone (O3) and nitrogen dioxide (NO2) in downtown Winnipeg increased over the study period and asthma prevalence increased twice as fast in Winnipeg as in the rest of the province. For Manitoba, the increase in preschool-aged asthma does not appear to be due to increased use of medical services, a change in ICD coding, an increase in the severity of the cases, or a decrease in income levels. The increases appear to be at least partly due to changes in diagnostic practices. The relationship between asthma and air pollution needs more detailed study as pollution is likely to be an important factor, particularly during the spring. Other areas for further investigation are changes in allergy and virus precursors, maternal smoking, and increased levels of pollens, molds and dust mites.

A case control study of cancer cervix patients attending Command Hospital, Pune.

A case control study was undertaken with the objective to determine the association of certain host factors like marital status, age at first marriage, duration of married life, parity, literary status and genital hygiene, in the subsequent development of cancer cervix. A total number of ninety-two cases and an equal number of control subjects, under matched case control design, were studied at Command Hospital, Pune over a period of nine months. Salient findings of this epidemiological study have been highlighted in this communication. Few of the risk factors viz early age at first marriage, longer duration of married life, increased and early parity, low educational status and poor genital hygiene were found to have played significant role in the subsequent development of carcinoma cervix.

Fluidity-dependent Mg2(+)-ATPase activity in membranes from Leishmania donovani promastigotes.

The state of the lipid phase of the membrane plays a key role in the exposure of various receptors, antigens and enzymes on the membrane surface. The fluidity of membranes of Leishmania donovani promastigotes was monitored by two independent methods, i.e. influx of sterol from liposomes and removal of phospholipids by treatment with phospholipase C. The altered sterol/phospholipid ratio, in both cases, provided evidence that the activity of the functionally important membrane-bound enzyme Mg2(+)-ATPase is modulated by the state of the lipid phase of the membrane.