Molecular characterization and serodiagnostic evaluation of the Echinococcus ortleppi recombinant glutaredoxin 1 protein for cystic echinococcosis in buffalo (Bubalus bubalis).

Cystic echinococcosis (CE), caused by the metacestode of Echinococcus granulosus sensu lato (s.l.), adversely affects the physiology of the vital organs in which they grow. Condemnation of meat causes substantial economic loss to the livestock industry. Conventionally the infection is detected by necropsy as serological diagnosis of the infection in livestock is ambiguous. Identification of specific diagnostic antigens would be a substitute for the cyst fluid antigens which lack adequate diagnostic sensitivity and specificity. BLAST analysis supported by the negligible pairwise nucleotide distance of the 389 nt COX1, 489 nt NAD1, and 425 nt ITS1 with the related sequences of E. ortleppi ascertained the association of E. ortleppi with CE in buffaloes. Given the extensive distribution of glutaredoxin 1 in every developmental stage of Echinococcus granulosus s.l that makes it an ideal serodiagnostic antigen for CE, we expressed the 14 kDa E. ortleppi glutaredoxin 1 (rEoGrx1) protein in E. coli BL21 (DE3) and tested a total of 225 sera samples, including 126 sera samples from the necropsy-positive buffalo, by the rEoGrx1 IgG-ELISA. The ELISA could detect a total of 82/126 sera samples as positive. The diagnostic sensitivity and specificity of the rEoGrx1 IgG-ELISA were 65.1 % and 51.5 %, respectively. The protein showed serological cross-reaction against Fasciola gigantica, Toxoplasma gondii, and Sarcocystis sp. The in silico bioinformatics analysis of the E. ortleppi, F. gigantica, and T. gondii glutaredoxin sequences revealed fully conserved amino acids at positions 11 and 21, the substitution of conserved amino acids at positions 14 and 6, and semi-conserved substitutions at positions 3 and 4, respectively. The findings partly explain the molecular basis of the serological cross-reactivity of the protein.

Universal Hearing Screening in Newborns Using Otoacoustic Emissions and Brainstem Evoked Response in Eastern Uttar Pradesh.

The objectives were to determine the incidence of hearing impairment in a standardized population of neonates and to determine the significance of association of epidemiological and risk factors with neonatal hearing loss. A cohort of 600 newborns was selected for study and divided into two groups-525 in 'No Risk' group and remaining 75 in 'At Risk' group. The study protocol was carried out in three steps: (a) Screening of Hearing Loss with TOAE, done from 36 h after birth to 28 days of life, (b) Re-screening of hearing loss in newborns (of 4-12 weeks of age), who were tested positive for hearing loss in the first screening, done with DPOAE, (c) Confirmation of hearing loss with BERA, in those who were tested positive in both the first and second screening. In the study the incidence of hearing impairment in 600 infants screened was 6.67 per 1000 screened; 3.81 per 1000 screened in the Not at Risk group and 26.67 per 1000 screened in At Risk group. In At Risk group, admitted to the NICU, severe birth asphyxia and hyperbilirubinemia were found to be major risk factors. Loss to follow up was more in Not at Risk group and False Positive cases with TEOAE were more than DPOAE. BERA was found to be must for confirmation of hearing loss. Neonatal Hearing Screening of only At Risk population is likely to miss some hearing loss. Universal Hearing Screening should be the preferred strategy. Good follow up in the 'At Risk' group suggests that initial interventions in NICU had sensitized the parents for the possibility of hearing loss. This study recommends the introduction of two stage neonatal screening-rescreening protocol, using OAE and BERA, in the country in phased manner.

Regulation of alternative splicing of Bcl-x by BC200 contributes to breast cancer pathogenesis.

BC200 is a long non-coding RNA (lncRNA) that has been implicated in the regulation of protein synthesis, yet whether dysregulation of BC200 contributes to the pathogenesis of human diseases remains elusive. In this study, we show that BC200 is upregulated in breast cancer; among breast tumor specimens there is a higher level of BC200 in estrogen receptor (ER) positive than in ER-negative tumors. Further experiments show that activation of estrogen signaling induces expression of BC200. To determine the significance of ER-regulated BC200 expression, we knockout (KO) BC200 by CRISPR/Cas9. BC200 KO suppresses tumor cell growth in vitro and in vivo by expression of the pro-apoptotic Bcl-xS isoform. Mechanistically, BC200 contains a 17-nucleotide sequence complementary to Bcl-x pre-mRNA, which may facilitate its binding to Bcl-x pre-mRNA and recruitment of heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1, a known splicing factor. Consequently, hnRNP A2/B1 interferes with association of Bcl-x pre-mRNA with the Bcl-xS-promoting factor Sam68, leading to a blockade of Bcl-xS expression. Together, these results suggest that BC200 plays an oncogenic role in breast cancer. Thus, BC200 may serve as a prognostic marker and possible target for attenuating deregulated cell proliferation in estrogen-dependent breast cancer.

Regulation of breast tumorigenesis through acid sensors.

The low extracellular pH in the microenvironment has been shown to promote tumor growth and metastasis; however, the underlying mechanism is poorly understood. Particularly, little is known how the tumor cell senses the acidic signal to activate the acidosis-mediated signaling. In this study, we show that breast cancer cells express acid-sensing ion channel 1 (ASIC1), a proton-gated cation channel primarily expressed in the nervous system. RNA interference, knockout and rescue experiments demonstrate a critical role for ASIC1 in acidosis-induced reactive oxidative species and NF-κB activation, two key events for tumorigenesis. Mechanistically, ASIC1 is required for acidosis-mediated signaling through calcium influx. We show that as a cytoplasmic membrane protein, ASIC1 is also associated with mitochondria, suggesting that ASIC1 may regulate mitochondrial calcium influx. Importantly, interrogation of the Cancer Genome Atlas breast invasive carcinoma data set indicates that alterations of ASIC1 alone or combined with other 4 ASIC genes are significantly correlated with poor patient survival. Furthermore, ASIC1 inhibitors cause a significant reduction of tumor growth and tumor load. Together, these results suggest that ASIC1 contributes to breast cancer pathogenesis in response to acidic tumor microenvironments, and ASIC1 may serve as a prognostic marker and a therapeutic target for breast cancer.

Characterization of excretory-secretory antigens of adult Toxocara canis by western blotting.

Toxocara canis is one of the most common helminth worm of dogs which continues to stimulate both public health concern alongside the higher scientific interest. It may cause visceral and ocular damage in humans especially in children. The identification of specific antigens of T. canis is important so as to develop better diagnostic techniques. Excretory-secretory (ES) antigens were prepared by culturing the adult T. canis worms in RPMI 1640 medium without serum supplementation followed by ammonium sulphate precipitation. These antigens were separated using sodium dodecyl sulphate-electrophoresis (SDS-PAGE). Recovered proteins ranged from 30 to 384 kDa. The specific reactivity of the T. canis excretory-secretory (TC-ES) proteins was checked by western blotting. The immuno-reactivity of the naturally infected dog sera with the TC-ES antigens showed five bands at 43, 57,105, 139 and 175 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against TC-ES antigens was observed with ten polypeptides of 21, 25, 30, 37, 45, 50, 57, 69, 77 and 105 kDa. Common antigens band were observed at 57 and 105 KDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocariasis in humans and adult dogs.

Evaluation of the effectiveness of diagnostic & management decision by teleophthalmology using indigenous equipment in comparison with in-clinic assessment of patients.

BACKGROUND & OBJECTIVES: There is a concern on the quality and the usefulness of teleophthalmology images, particularly those using indigenous equipment, in making a diagnosis and treatment decisions in ophthalmology. The present study was done to compare the level of agreement and sensitivity and specificity of diagnosis and management decisions of various eye diseases by teleophthalmology using indigenous equipment, compared to the in-clinic assessment. METHODS: Patients having different eye diseases were evaluated by two ophthalmologists - one ophthalmologist examined the patient in clinic setting while the other ophthalmologist made the diagnosis and management decision based on images sent by teleophthalmology. The images were taken by the ophthalmic technician using digital imaging system and fundus camera. The clinical findings and management decisions by the two ophthalmologists were masked to each others. RESULTS: In diagnosis of anterior segment eye diseases such as cataract and corneal diseases there was good to very good agreement (kappa values of 0.68 and 0.91 for cataract and corneal diseases respectively) between in-clinic assessment and assessment by teleophthalmology. There was moderate agreement (kappa values of 0.52 and 0.48 for glaucoma and retinal diseases respectively) between in-clinic assessment and assessment by teleophthalmology for the diagnosis of glaucoma and retinal diseases. For the management decisions of patients, there was moderate level of agreement in all groups of eye diseases. INTERPRETATION & CONCLUSIONS: Teleophthalmology, using indigenous equipment was found to be effective in diagnosis and management decision of anterior segment eye diseases such as cataract and cornea, and with some modification and continuous training to the technicians could become an effective tool for screening and referral of glaucoma and retinal diseases.

Pentoxifylline therapy in the management of oral submucous fibrosis.

OBJECTIVES: Oral submucous fibrosis is a common premalignant condition in the Indian subcontinent and is caused by chewing areca nut and other irritants in various forms. Its medical treatment is not yet fully standardized. In this study we compared the efficacy of Pentoxifylline as compared to placebo. MATERIALS AND METHODS: 75 patients suffering from oral submucous fibrosis were randomly divided into two groups A and B. Group A patients received placebo, while Group B patients received 400 mg. Pentoxifylline for a period of 7 months. Treatment outcome was evaluated on the basis of improvement in symptom and sign scores. Student's t test was applied for comparing the results. RESULTS: The improvement in total (i.e. symptoms + sign) score was 25% in group A and 49.15% in group B. This difference was found to be statistically significant. (p < 0.05) CONCLUSION: Treatment regimen of group B was more effective. No significant side effects were seen. A follow up study is required to assess long term outcome of this therapy.

Effect of cigarette smoking on the periodontal health status: A comparative, cross sectional study.

CONTEXT: Survey. AIMS: The objective of the study was to evaluate the periodontal health status among cigarette smokers and non cigarette smokers, and oral hygiene measures. SETTINGS AND DESIGN: Cross sectional study. MATERIALS AND METHODS: The study included 400 male (200 cigarette smokers and 200 non smokers) aged 18-65 years. The subjects were randomly selected from the patients attending dental out-patient department of civil hospital and Himachal Dental College, Sundernagar. Community Periodontal Index (CPI) score was recorded for each patient and a questionnaire was completed by each patient. STATISTICAL ANALYSIS USED: Chi square and t-test. RESULTS: Periodontal condition as assessed by CPI score showed that there was statistically significant difference in the findings between cigarette smokers and non-smokers. CONCLUSIONS: Within the limits of this study, positive association was observed between periodontal disease and cigarette smoking. It was found that cigarette smoking was associated with lesser gingival bleeding and deeper pockets as compared to non-smokers.

Solitary nasal schwannoma clinically presenting as an angiofibroma of the nasopharynx.

Benign schwannomas of the nasal cavity are rare. When they do occur, their clinical presentation is variable, and some are fairly vascular. We report the case of a 17-year-old boy who presented with epistaxis associated with a unilateral nasal mass that extended into the nasopharynx. The tumor was removed via a transpalatal approach. In most such cases, a clinical diagnosis of angiofibroma is made. In this case, however, histopathologic examination identified the lesion as a schwannoma.

Efficacy of hydrocortisone acetate/hyaluronidase vs triamcinolone acetonide/hyaluronidase in the treatment of oral submucous fibrosis.

BACKGROUND & OBJECTIVES: Oral submucous fibrosis is a common premalignant condition caused by chewing arecanut and other irritants in various forms. Its medical treatment is not yet fully standardized, although the optimal doses of its medical treatment is in the form of hydrocortisone acetate combined with hyaluronidase. The problem with the prevailing treatment was injections at weekly interval. In this study we compared the efficacy of hydrocortisone acetate and hyaluronidase at weekly interval versus triamcinolone acetonide and hyaluronidase at 15 days interval. METHODS: Patients of OSMF (100) were randomly divided into two groups A and B. Group A patients received combination of hydrocortisone acetate (1.5 ml)/hyaluronidase (1500 IU) at weekly interval submucosally in pterygomandibular raphe, half dose on each side for 22 wk. Group B patients received combination of triamcinolone acetonide (10 mg/ml)/ hyaluronidase (1500 IU) at 15 days interval for 22 wk. Treatment outcome was evaluated on the basis of improvement in symptom score, sign score and histopathological improvement. Student's 't' test was applied for comparing the results. RESULTS: No statistically significant difference in symptom score, sign score and histopathological improvement was seen between the two groups. INTERPRETATION & CONCLUSION: Treatment regimen of group B was more convenient to the patients because less number of visits required and cheap. No side effects were seen. A follow up study is required to see long term effects.

Role of measurement of nasal mucociliary clearance by saccharine test as a yard stick of success of functional endoscopic sinus surgery.

AIM: The present study was done to establish normal nasal mucociliary clearance time (NMCT) in people of our region, study variations in NMCT in patients of chronic rhinosinusitis, to see change in NMCT after functional endoscopic sinus surgery (FESS) and to see whether saccharine test can be used as a yard stick of success of FESS. MATERIALS AND METHODS: NMCT of 100 normal individual was measured twice to establish control value. Fourtythree patients of chronic rhinosinusitis were divided into polypoidal and non-polypoidal. Their symptom score, endoscopic appearance score, CT scan score and NMCT was measured preoperatively and postoperatively. RESULTS: NMCT in control group was 6.61 ± 0.84 minutes, in unilateral (u/l) polypoidal sinusitis 13.45 ± 2.07 minutes, in bilateral (b/l) polypoidal sinusitis 21.31 ± 0.76 minutes, u/l non-polypoidal sinusitis 9.54 ± 1.00 minutes and in b/l non-polypoidal sinusitis 11.34 ± 0.93 minutes. Linear relationship was seen between preoperative NMCT and preoperative symptom score, endoscopic appearance score, CT scan score. NMCT improved significantly after FESS. CONCLUSION: NMCT measurement helps in selection of appropriate patients for surgery. It can be used as preoperative indicator of success of FESS.

PLGA nanoparticles stabilized with cationic surfactant: safety studies and application in oral delivery of paclitaxel to treat chemical-induced breast cancer in rat.

PURPOSE: This study was carried out to formulate poly(lactide-co-glycolide) (PLGA) nanoparticles using a quaternary ammonium salt didodecyl dimethylammonium bromide (DMAB) and checking their utility to deliver paclitaxel by oral route. METHODS: Particles were prepared by emulsion solvent diffusion evaporation method. DMAB and particles stabilized with it were evaluated by MTT and LDH cytotoxicity assays. Paclitaxel was encapsulated in these nanoparticles and evaluated in a chemical carcinogenesis model in Sprague Dawley rats. RESULTS: MTT and LDH assays showed the surfactant to be safe to in vitro cell cultures at concentrations <33 microM. PLGA nanoparticles prepared using this stabilizer were also found to be non-toxic to cell lines for the duration of the study. When administered orally to rats bearing chemically induced breast cancer, nanoparticles were equally effective/better than intravenous paclitaxel in cremophor EL at 50% lower dose. CONCLUSIONS: This study proves the safety and utility of DMAB in stabilizing preformed polymers like PLGA resulting in nanoparticles. This preliminary data provides a proof of concept of enabling oral chemotherapy by efficacy enhancement for paclitaxel.

Lipid profile in oral submucous fibrosis.

BACKGROUND: Changes in lipid profile have long been associated with malignancies as lipids play a key role in maintenance of cell integrity. This study evaluated the alterations in extended lipid profile in untreated patients of oral submucous fibrosis (OSMF) and studied the correlation between lipid levels with tobacco consumption. MATERIALS AND METHODS: In this hospital-based study, 65 clinically diagnosed and histopathologically proven patients of OSMF and 42 age and sex matched controls were studied. In these samples serum lipids including: (i) Total cholesterol, (ii) LDL cholesterol (LDLC), (iii) HDL cholesterol (HDLC) (iv) VLDL cholesterol (VLDLC) (v) triglycerides (vi) Apo-A1 (viii) Apo-B and (viii) LPa were analyzed. RESULTS: A significant decrease in plasma total cholesterol, HDLC and Apo-A1 was observed in patients with OSMF as compared to the controls. Thus an inverse relationship between plasma lipid levels and patients was found in OSMF. CONCLUSION: The lower levels of plasma cholesterol and other lipid constituents in patients might be due to their increased utilization. The findings strongly warrant an in-depth study of alterations in plasma lipid profile in patients with oral precancerous conditions.

Improvement of in vitro oocyte maturation with lectin supplementation and expression analysis of Cx43, GDF-9, FGF-4 and Fibronectin mRNA transcripts in Buffalo (Bubalus bubalis).

PURPOSE: To optimize In vitro maturation (IVM) of quality oocytes for embryo production through IVF and SCNT. METHODS: Buffalo oocytes were in vitro matured in the presence of the pokeweed lectin (Phytolacca americana), a potent lymphocyte mitogen. Lectin was supplemented in TCM + 10% FBS at the doses of 0, 1, 5, 10, 15, 20 and 40 microg/ml and cumulus expansion and gene expression patterns were characterized. RESULTS: The degree of cumulus expansion in different lectin treatment levels improved from 1.1 at 1 Ag/ml level to 3.60 at 10 microg/ml level and then decreased in higher concentration 20 microg/ml (1.66) and 40 microg/ml (0.64). IVF embryos showed highest cleavage rate (88.8%) in 10 microg/ml lectin treatment. Expression of all mRNA transcript studied (Cx43, GDF 9, FGF-4 and Fibronectin) was positively correlated with cumulus expansion and polar body extrusion. CONCLUSIONS: Mitogenic lectin supplemented maturation media improves oocyte quality for in vitro embryo production.

Assessment of deafmute patients: a study of ten years.

Present prospective study was conducted on 350 patients presenting with chief complaints of impaired hearing and delayed speech from 1996-2006. The aim of the present study was to find out the prevalence of deafmutism in our area, the aetiology of childhood deafness and to ascertain the role of acoustic reflex test (ART) for hearing screening considering brain stem evoked response audiometry (BSERA) as gold standard. A detailed history, clinical and other relevant systemic examination and investigations were done to find out the cause. All patients were subjected to ART test and BSERA. Male to female ratio was 2.1:1. Prevalence was found to be 5.59 per one lac population in our district. The commonest age of presentation was in the second decade. The causes for childhood deafness were genetic (15.8%), embryopathies (10%), perinatal (10.8%) and postnatal problems (12.5%). In 50.6% cases it was aediopathic. Congenital syndromic abnormalities were found in 5.4% patients. In BSERA 21% patients were found to have residual hearing varying from 60 dB to 100dB. The positive predictive value of ART was found to be low (10.4%). Hence it was recommended that ART should not be used as screening tool for childhood deafness.

Our experiences with parapharyngeal space tumors and systematic review of the literature.

OBJECTIVES: To find out the geographical distribution, age, sex, mode of presentation, diagnostic modalities, surgical approaches, postoperative complications, follow-up and histopathology of parapharyngeal space tumors (PFT) through world wide web search (WWWS) and PFT of our series. MATERIALS AND METHODS: A retrospective study of all PFT seen and treated in S.R.N Hospital, Allahabad between 1990 and 2006 was done. A systematic review of the literature was done through WWWS. RESULTS: PFT were commonest in the Asian zone. Incidence was highest in the 3rd and 4th decade. Oropharyngeal (100%) and cervical (91.6%) swelling were the commonest presentation. MRI is better than CT scan in evaluating fat plane. Transcervical approach was the commonest. In our series, benign tumors (83.4%) were common, constituting 50% of neurogenic and 16.6% of salivary gland origin. CONCLUSION: PFT were more prevalent in the Asian zone. Neurogenic tumors were commonest in our study which contradicts the WWWS of having maximum PFT of salivary gland in origin.

Effects of radiotherapy on parotid salivary sialochemistry in head and neck cancer patients.

AIM: To determine the effects of high dose irradiation on parotid salivary sodium and pH concentration at subsequent duration of 1.5, 3 and 6 months following radiotherapy. MATERIALS AND METHODS: Eighty parotid glands of head and neck cancer patients were irradiated with mean dose of 66 Gy. The stimulated parotid flow (PF) was collected by a cannulation of Stenson's duct followed by analysis of sodium (PF sodium) by Easylyte Sodium/Potassium auto analyzer and pH by litmus narrow band pH paper. RESULTS: A steep elevation of PF sodium was found in post-RT period after 1.5 months of starting RT followed by gradual increase up to 6 months and pH changed towards acidity. CONCLUSIONS: A high dose of 66 Gy causes irreversible damage to parotid salivary duct system.

Effectiveness of red cell osmotic fragility test with varying degrees of saline concentration in detecting beta-thalassaemia trait.

INTRODUCTION: This study was performed with the objective of determining the efficacy of naked eye single tube red cell osmotic fragility test (NESTROFT) as a screening test for beta-thalassaemia trait, and to standardise a saline concentration which could give best results with minimum error and maximum sensitivity and specificity. METHODS: Five concentrations (0.35 percent, 0.36 percent, 0.37 percent, 0.38 percent and 0.39 percent) of buffered saline solutions were used. NESTROFT was applied to three groups of subjects: 24 normal individuals, 87 subjects with genetically-proven beta-thalassaemia trait and 13 patients with proven iron-deficiency anaemia. RESULTS: The results demonstrated that 0.36 percent was the best saline concentration for NESTROFT. It could detect 97.7 percent of heterozygous beta-thalassaemia patients, compared to 94.25 percent, 91.95 percent, 88.51 percent and 82.76 percent detection rates obtained with 0.35 percent, 0.37 percent, 0.38 percent and 0.39 percent saline concentrations, respectively. Specificity of NESTROFT with 0.36 percent saline was also higher at 83.3 percent, whereas that of 0.35 percent, 0.37 percent, 0.38 percent and 0.39 percent was 79.17 percent, 79.17 percent, 70.83 percent and 62.5 percent, respectively. This test with 0.36 percent saline concentration was also positive for three (23.08 percent) patients with iron-deficiency anaemia. CONCLUSION: NESTROFT done with 0.36 percent buffered saline solution provides more accurate results compared to the other concentrations tested. Since the test is sensitive, cost-effective, rapid and reliable, it may be considered as the single screening test to be used in areas with limited laboratory facilities and economic resources.