Application of vaginal tampon as an alternative to nasal swabs for higher recovery of DNA from sheep and goats for PCR based diagnosis of bovine tuberculosis.

A study was conducted to find the applicability of vaginal tampons as an alternative to regular cotton swabs as a nasal secretion collection tool for the higher recovery of DNA. Nasal secretions were collected from sheep and goats using regular cotton swab and tampon swab. The mean yield and purity of the DNA extracted from tampon were significantly higher than that of the DNA extracted from cotton swab. The tampon swabs resulted higher DNA recovery than the cotton swabs after they were allowed to absorb M. bovis culture. The tampon swab was also found to be more sensitive in detecting M. bovis by PCR. This study concluded that vaginal tampons are having a higher absorption capacity with more DNA yield and can be used as a nasal swab in the diagnosis of bovine tuberculosis.

Tumor preventive and antioxidant efficacy of chlorogenic acid-loaded chitosan nanoparticles in experimental skin carcinogenesis.

Oxidative stress, a pathological condition, contributes to the pathophysiology of a number of diseases including carcinogenesis. Numerous studies pointed out the disturbed antioxidants status and accumulation of oxidative stress markers in the carcinogenesis. The present study analyzed the anticancer efficacy of chlorogenic acid-loaded chitosan nanoparticles by utilizing the oxidative stress biomarkers as an endpoint in mice with skin cancer developed by 7,12-dimethylbenz(a)anthracene (DMBA). Oxidative stress markers' (lipid peroxidation by-products and antioxidants) levels or activities were measured using colorimetric assays. While mice exposed with DMBA alone showed a 100% tumor incidence, 0 and 50% tumor formation was seen in mice treated with DMBA + topical application of the nanoparticles and DMBA + orally administered nanoparticles, respectively. Also, the study noticed a 33% and 67% tumor incidence in mice treated with DMBA + topical application of free chlorogenic acid and DMBA + orally administered free chlorogenic acid, respectively. The present study noticed that the topical application of chlorogenic acid-loaded chitosan nanoparticles to DMBA-painted mice completely suppressed the tumor growth and restored the levels or activities of oxidative stress markers as compared to mice that received DMBA + oral administration of chlorogenic acid-loaded chitosan nanoparticles. The study observed that chlorogenic acid-loaded chitosan nanoparticles are more potent than free chlorogenic acid in preventing skin cancer in mice caused by DMBA. Thus, the present investigation explores the tumor-inhibiting efficacy of chlorogenic acid-loaded chitosan nanoparticles in experimental skin cancer, and the tumor preventive efficiency could be attributed to their antilipid peroxidative and antioxidant effects.

A Novel CNN-TLSTM Approach for Dengue Disease Identification and Prevention using IoT-Fog Cloud Architecture.

One of the mosquito-borne pandemic viral infections is Dengue which is mostly transmitted to humans by the Aedes agypti or female Aedes albopictis mosquitoes. The dengue disease expansion is mainly due to the different factors such as climate change, socioeconomic factors, viral evolution, globalization, etc. The unavailability of certain antiviral therapy and specific vaccine increases the risk of the dengue disease spreading even further. This arises the need for a novel technique that overcomes the complexities associated with dengue disease prediction such as low reporting level, misclassification, and incompatible disease monitoring framework. This paper mainly overcomes the above-mentioned problems by integrating the Internet of Things (IoT), fog-cloud, and deep learning techniques for efficient dengue monitoring. A compatible disease monitoring framework is formed via the IoT devices and the reports are effectively created and transferred to the healthcare facilities via the fog-cloud model. The misdiagnosis error is overcome in this paper using the novel Hybrid Convolutional Neural Network (CNN) with Tanh Long and Short Term Memory (TLSTM) based Adaptive Teaching Learning Based Optimization (ATLBO) algorithm. The ATLBO optimized CNN-TLSTM architecture mainly analyzes the dengue-related parameters such as Soft Bleeding, Muscle Pain, Joint Pain, Skin rash, Fever, Water Site, Carbon Dioxide, Water Site Humidity, Water Site Temperature, etc. for an efficient clinical decision making and timely disease diagnosis. The experimental results are conducted using a real-time dataset and its performance is validated using various performance metrics. When compared in terms of different statistical parameters such as accuracy, f-measure, mean square error, and reliability, the proposed method offers superior results in the case of dengue disease detection than other existing methods. The ATLBO optimized hybrid CNN-TLSTM shows an accuracy of 96.9%, a precision of 95.7%, recall of 96.8%, and F-measure of 96.2% which is relatively high when compared to the existing techniques. The proposed model is capable of identifying the patients in a certain geographical region and preventing the disease emergency via immediate disease diagnosis and alerting the healthcare officials to offer the stipulated services.

Diagnosing Breast Cancer Based on the Adaptive Neuro-Fuzzy Inference System.

In this work, a novel hybrid neuro-fuzzy classifier (HNFC) technique is proposed for producing more accuracy in input data classification. The inputs are fuzzified using a generalized membership function. The fuzzification matrix helps to create connectivity between input pattern and degree of membership to various classes in the dataset. According to that, the classification process is performed for the input data. This novel method is applied for ten number of benchmark datasets. During preprocessing, the missing data is replaced with the mean value. Then, the statistical correlation is applied for selecting the important features from the dataset. After applying a data transformation technique, the values normalized. Initially, fuzzy logic has been applied for the input dataset; then, the neural network is applied to measure the performance. The result of the proposed method is evaluated with supervised classification techniques such as radial basis function neural network (RBFNN) and adaptive neuro-fuzzy inference system (ANFIS). Classifier performance is evaluated by measures like accuracy and error rate. From the investigation, the proposed approach provided 86.2% of classification accuracy for the breast cancer dataset compared to other two approaches.

An Expert System for COVID-19 Infection Tracking in Lungs Using Image Processing and Deep Learning Techniques.

The proposed method introduces algorithms for the preprocessing of normal, COVID-19, and pneumonia X-ray lung images which promote the accuracy of classification when compared with raw (unprocessed) X-ray lung images. Preprocessing of an image improves the quality of an image increasing the intersection over union scores in segmentation of lungs from the X-ray images. The authors have implemented an efficient preprocessing and classification technique for respiratory disease detection. In this proposed method, the histogram of oriented gradients (HOG) algorithm, Haar transform (Haar), and local binary pattern (LBP) algorithm were applied on lung X-ray images to extract the best features and segment the left lung and right lung. The segmentation of lungs from the X-ray can improve the accuracy of results in COVID-19 detection algorithms or any machine/deep learning techniques. The segmented lungs are validated over intersection over union scores to compare the algorithms. The preprocessed X-ray image results in better accuracy in classification for all three classes (normal/COVID-19/pneumonia) than unprocessed raw images. VGGNet, AlexNet, Resnet, and the proposed deep neural network were implemented for the classification of respiratory diseases. Among these architectures, the proposed deep neural network outperformed the other models with better classification accuracy.

Diagnosis of Cervical Cancer based on Ensemble Deep Learning Network using Colposcopy Images.

Traditional screening of cervical cancer type classification majorly depends on the pathologist's experience, which also has less accuracy. Colposcopy is a critical component of cervical cancer prevention. In conjunction with precancer screening and treatment, colposcopy has played an essential role in lowering the incidence and mortality from cervical cancer over the last 50 years. However, due to the increase in workload, vision screening causes misdiagnosis and low diagnostic efficiency. Medical image processing using the convolutional neural network (CNN) model shows its superiority for the classification of cervical cancer type in the field of deep learning. This paper proposes two deep learning CNN architectures to detect cervical cancer using the colposcopy images; one is the VGG19 (TL) model, and the other is CYENET. In the CNN architecture, VGG19 is adopted as a transfer learning for the studies. A new model is developed and termed as the Colposcopy Ensemble Network (CYENET) to classify cervical cancers from colposcopy images automatically. The accuracy, specificity, and sensitivity are estimated for the developed model. The classification accuracy for VGG19 was 73.3%. Relatively satisfied results are obtained for VGG19 (TL). From the kappa score of the VGG19 model, we can interpret that it comes under the category of moderate classification. The experimental results show that the proposed CYENET exhibited high sensitivity, specificity, and kappa scores of 92.4%, 96.2%, and 88%, respectively. The classification accuracy of the CYENET model is improved as 92.3%, which is 19% higher than the VGG19 (TL) model.

Health and quality of life among a cohort of patients having lateral internal sphincterotomy for anal fissures.

AIM: The aim of this study is to report changes in health-related quality of life attributable to lateral internal sphincterotomy for treatment of anal fissure. There is very little evidence on whether the overall health-related quality of life of patients is detrimentally affected by the condition, or which aspects of self-perceived health status improve after lateral internal sphincterotomy. This study will articulate which aspects of health tend to improve and guide postoperative expectations appropriately. Knowledge gained from this study may also identify gaps in an individual patient's episode of care. METHOD: Patients were prospectively identified when they consented to surgical treatment of their anal fissure and were contacted by phone to participate. Participants completed a number of patient-reported outcomes preoperatively and 6 months postoperatively. Faecal incontinence-related quality of life, pain and depression were measured at both time points. The severity of faecal incontinence was measured at both times. RESULTS: Participants reported high levels of pain preoperatively. Postoperatively, improvement in pain exceeded the threshold of clinical relevance (P < 0.01). Thirty-five per cent of participants reported significant effects of faecal incontinence preoperatively, while 26% did so postoperatively. Participants with multiple comorbidities were more likely to report faecal incontinence postoperatively than preoperatively. CONCLUSION: This study reports that lateral internal sphincterotomy improved pain symptoms without adverse effects on continence. Not all domains of health-related quality of life were similarly positively affected by anal fissure repair.

Duck plague outbreak in a Chara-Chemballi duck farm.

BACKGROUND: Duck rearing is one of the important livelihoods of rural people. Duck plague is one of the diseases causing heavy mortality resulting in economic losses. CASE DESCRIPTION: An outbreak of duck plague in a farm in Kadavakathi Village near Tenkasi, Tirunelveli Dt., is reported. FINDINGS/TREATMENT AND OUTCOME: Two thousands out of 4500 Chara-Chemballi breed of ducks which were recently purchased from Chenganacherry in Kerala died, with a mortality rate of 44.4%. Clinical signs of inappetence, partial closure of eyelid, conjunctivitis, corneal opacity, oculo-nasal discharge, soiled vent with green white watery diarrhoea, ataxia, incoordination and sudden death were observed. Necropsy examination revealed diphtheritic membrane in the oesophagus, congestion, petechial haemorrhages and multifocal gray white areas on the surface of the liver, epicardial haemorrhages, congested trachea, lung, kidneys, splenomegaly with mottled appearance and enteritis. Microscopical examination revealed presence of eosinophilic intranuclear and intracytoplasmic inclusions in the epithelial cells of the intestine and hepatocytes, degeneration and necrosis of enterocytes, dilated crypt epithelial cells with presence of eosinophilic intranuclear and intracytoplasmic inclusions, congestion and lymphoid cell depletion in the spleen, vasculitis, congestion, and haemorrhages in the trachea and lungs, proventriculitis, and congested kidneys. Polymerase chain reaction (PCR) also confirmed the duck plague viral infection by the amplification of polymerase gene fragment (446 bp). CONCLUSION: Based on the above findings, the Chara-Chemballi duck disease outbreak was diagnosed as duck viral enteritis infection.

Two-stage hypospadias repair with a free graft for severe primary and revision hypospadias: A single surgeon's experience with long-term follow-up.

INTRODUCTION: Repair of severe primary and revision hypospadias is a demanding procedure. Debate continues as to whether a two-stage approach or single-stage technique is superior. The two-stage procedure with a free graft involves penile straightening followed by application of a graft for the neourethral plate at stage one; with tubularization at stage two after graft maturation. OBJECTIVE: To report the outcomes of a single surgeon's experience with the two-stage repair using a free graft for both severe primary and revision hypospadias with long-term follow-up. MATERIALS AND METHODS: Between July 1998 and January 2010, 301 boys underwent a two-stage reconstruction. The surgical technique is described in the manuscript. Primary repairs (n = 208): indications for a two-stage approach with a free graft included meatal position, presence of corporal chordee, and poor glans development. Median follow-up from completion of the second stage was 75 months. Revision repairs (n = 93): indications included urethral fistula, excessive scarring/meatal stenosis, balanitis xerotica obliterans (BXO), and residual or untreated chordee. Median follow-up from completion of the second stage was 85 months. RESULTS: For the primary repairs (n = 208), the graft took well in all but one case. Second-stage complications included fistula (7), meatal stenosis (3), partial glans dehiscence (3), and all were re-operated (13). For the revision repairs (n = 93), the graft took well in all but four cases. Second-stage complications included fistula (5), meatal stenosis (3), breakdown (1) and reoperation (8). DISCUSSION: In a systematic review of 20 years of publications on the repair of primary severe hypospadias, the two-stage procedure with a free graft demonstrated an overall complication rate of 22%; this was a distinct overall benefit when compared with the single-stage procedures in terms of lower complication rates (Castagnetti and El-Ghoneimi, 2010). Our results for the severe primary repairs revealed significantly lower complication rates than those in the literature, with an overall re-operation rate of 6.3%, a fistula rate of 3.4%, and meatal stenosis and partial glans dehiscence at 1.4% each. Several papers have documented outcomes following the single-stage tubularized incised plate urethroplasty for re-operative hypospadias, giving overall complication rates ranging from 15.4 to 30%. Our data show a re-operative rate of 8.6%, a fistula rate of 5.3%, breakdown in 1.1%, and meatal stenosis in 3.2%. CONCLUSION: The two-stage repair with a free graft for correction of both severe primary and failed primary hypospadias is a safe, viable, and durable procedure offering low morbidity and excellent cosmetic results. The authors advocate the two-stage repair with a free graft as the technique of choice for treatment of both of these challenging groups of the deformity.

Alzheimer's Disease and Medicinal Plants: An Overview.

Alzheimer's disease (AD) is progressive neurodegenerative disorder and identified as a major health concern globally. Individuals with AD and their care givers are affected in personal, emotional, financial, and social levels. Due to its significant impact and heavy burden on the individual, the patients' families, and society, it is highly needed to search for cost effective, long-time retention therapeutic targets. In recent decades, there are lots of research conducted the possible benefit of natural products and their active components on AD and other neurodegenerative disease, which are discussed here.

Testicular atrophy following paediatric primary orchidopexy: A prospective study.

BACKGROUND: With the Nordic consensus statement advocating orchidopexy at an earlier age, the present study sought to investigate the outcomes of primary paediatric orchidopexy at a tertiary UK centre. OBJECTIVE: To prospectively assess testicular atrophy following primary orchidopexy for undescended testes in a paediatric population. Secondary outcomes were complication rates and whether outcomes were dependent on grade of operating surgeon. STUDY DESIGN: Prospective data regarding age at operation, classification of the undescended testis, length of follow-up, and subjective comparison of intraoperative and postoperative testicular volumes compared with the contralateral testis were collected. Testicular atrophy was defined as >50% loss of testicular volume or a postoperative testicular volume <25% of the volume of the contralateral testis. Patients were excluded for incomplete data and follow-up <6 months. RESULTS: Data for 234 patients were analysed. Testicular atrophy occurred in 2.6% of cases. There was no reported testicular re-ascent. All secondary acquired cases underwent a previous ipsilateral hernia repair. There was no significant difference in outcomes comparing the grade of surgeon (consultant n = 8, trainee/staff-grade surgeon n = 7-8). There was a trend towards postoperative catch-up growth in approximately one fifth of cases. DISCUSSION: Previous studies have reported a testicular atrophy rate of 5%. The present study reported a similar rate of 2.6%. In agreement with a previous publication, it was also found that testicular atrophy was not dependent on the grade of operating surgeon. The mechanism for testicular catch-up growth is not well understood. Animal studies have supported the hypothesis that increased temperature has a detrimental effect on testicular volume. However, follow-up in the present cohort was short (median 6.9 months), making interpretation of this finding difficult. It is acknowledged that clinical palpation alone to determine testicular volume potentially introduces intra-observer and inter-observer error. However, prospective studies using ultrasound to determine testicular volumes following orchidopexy have reported catch-up growth. CONCLUSION: This study represented one of the larger collections of prospective assessments of outcomes following primary orchidopexy. It was acknowledged that subjectively assessing testicular volume is not ideal; however, the data correlated with similar studies.

Thirty-day readmissions following adult spinal deformity surgery: an analysis of the National Surgical Quality Improvement Program (NSQIP) database.

BACKGROUND: Adult spine deformity surgery (ASDS) is a significantly invasive procedure with a relatively high complication rate. The thirty-day hospital readmission rate following surgery is an important quality measure monitored by multiple quality reporting agencies. PURPOSE: This study seeks to determine the risk factors for 30- day readmission rate in patients undergoing ASDS and identify the risk factors associated with readmission. STUDY DESIGN: This is a retrospective multicenter study. METHODS: The National Surgical Quality Improvement Program database, which is a large multi-institutional database, was searched for patients that underwent ASDS from 2011 to 2013. The patients were identified by searching seven Current Procedural Terminology codes most commonly used for spinal deformity surgery. Twenty-seven preoperative variables, including patient demographics and comorbidities, intraoperative parameters, and postoperative complications were analyzed to identify risk factors for readmission. RESULTS: A total of 747 adult patients who underwent ASDS were identified. Of the 747 patients, 7.5% (56/747) were readmitted within 30 days. The most common causes of readmission were infection (n=11), hematoma or seroma formation (n=5), and postoperative pain (n=3). Univariate analysis revealed male gender (p=.038, odds ratio [OR]=1.83) and pulmonary embolism before discharge (p=.048, OR=8.44) to be associated with readmission. In multivariate analysis, obesity (p=.047, OR=1.80), peripheral vascular disease (p=.045, OR=17.52), pulmonary embolism before discharge (p=.012, OR=10.35), and total or partial dependent preoperative functional health status (p=.041, OR=2.45), were found to be independent risk factors for readmission. Age, smoking, and resident involvement during surgical procedure were among the many factors not associated with increased risk of readmission. CONCLUSIONS: The 30-day readmission rate for ASDS is increasingly becoming a significant health-care quality indicator. Patients with the aforementioned significant risk factors should be closely followed up, which can potentially avoid subsequent readmission.

Enicostemma littorale prevents tumor formation in 7,12-dimethylbenz(a)anthracene-induced hamster buccal pouch carcinogenesis.

Oral cancer is one of the most common malignancies worldwide, and India has recorded the highest annual incidence of oral cancer in comparison with other countries. Altered lipid peroxidation and antioxidant status along with defect in detoxification cascade have been implicated in the pathogenesis of several cancers including oral cancer. The aim of this study was to investigate the chemopreventive potential of ethanolic extract of Enicostemma littorale leaves (ElELet) in 7,12-dimethylbenz(a)anthracene (DMBA)-induced hamster buccal pouch carcinogenesis. Oral tumor was developed in the buccal pouches of male golden Syrian hamsters by painting with 0.5% DMBA three times a week for 14 weeks. We observed 100% tumor formation with increase in tumor volume and tumor burden in the hamsters treated with DMBA alone. Imbalance in phase I (cytochrome P450 and cytochrome b5) and phase II (glutathione reductase, glutathione-S-transferase, glutathione, and Deoxythymidine-diaphorase (DT)-diaphorase) detoxification agents and lipid peroxidation by-products (thiobarbituric acid reactive substances) and antioxidant (superoxide dismutase, catalase, glutathione peroxidase, and vitamins E and C) status was noticed in hamsters treated with DMBA alone. Oral administration of ElELet at a dose of 250 mg/kg body weight to hamsters treated with DMBA significantly prevented both precancerous and cancerous lesions in the oral cavity. ElELet modulated the status of phase I and II detoxification agents and antioxidants in favor of the suppression of oral carcinogenesis. This study thus suggests that E. littorale might have inhibited the oral carcinogenesis in DMBA-treated hamsters through its antioxidant potential. The present findings are also substantiated by histological studies during DMBA-induced oral carcinogenesis.

WITHDRAWN:Development of Dot Enzyme Linked Immuno Sorbent Assay Using Recombinant Hexon Protein for Detecting Antibodies to Hydropericardium Syndrome Disease Virus.

Ahead of Print article withdrawn by publisher.

Co-culture: A quick approach for isolation of street rabies virus in murine neuroblastoma cells.

BACKGROUND: Laboratory detection of rabies in most cases is based on detection of the antigen by fluorescent antibody test, however, in weak positive cases confirmative laboratory diagnosis depends on widely accepted mouse inoculation test. Cell lines like neuroblastoma have been used to isolate the virus with greater success not only to target for diagnosis, but also for molecular studies that determine the epidemiology of the circulating street rabies strains and in studies that look at the efficiency of the developed monoclonal antibodies to neutralize the different rabies strains. Due to the recent issues in obtaining ethical permission for mouse experimentation, and also the passages required in the cell lines to isolate the virus, we report herewith a co-culture protocol using the murine neuroblastoma (MNA) cells, which enable quicker isolation of street rabies virus with minimum passages. OBJECTIVE: This study is not to have an alternative diagnostic assay, but an approach to produce sufficient amount of rabies virus in minimum passages by a co-culture approach in MNA cells. MATERIALS AND METHODS: The MNA cells are co-cultured by topping the normal cells with infected cells every 48 h and the infectivity was followed up by performing direct fluorescent-antibody test. RESULTS: The co-culture approach results in 100% infectivity and hence the use of live mouse for experimentation could be avoided. CONCLUSION: Co-culture method provides an alternative for the situations with limited sample volume and for the quicker isolation of virus which warrants the wild type strains without much modification.

Metastatic spinal cord compression as a result of the unknown primary tumour.

PURPOSE: Patients presenting with metastatic spinal cord compression (MSCC) due to an unknown primary tumour (UPT) present an interesting problem with limited literature available to provide guidance on management. Our aim was twofold-first, to analyse all our patients with MSCC due to a UPT pre-operatively, to review their treatment and outcome; second, make comparisons with those patients who presented with MSCC due to a known primary tumour (KPT) during the same period. METHODS: All data was collected retrospectively from October 2004 to October 2009, then prospectively from October 2009 to October 2012 (8 years). We reviewed all patient records held on the database, including patient demographics, primary tumour, neurological outcome (Frankel grade), complications and survival. RESULTS: During the 8-year study period, out of the 382 patients who underwent emergency surgery for MSCC, 285 patients were included in whom complete information was available. Of these, 17 patients presented with MSCC due to a UPT (6 %; mean age 61 years, 5 M, 12 F). When compared to those with a known primary, the UPT group trended to a longer duration of symptoms prior to surgery (200 vs. 156 days, p = 0.86). They had a similar neurological outcome (88 % remained the same or improved post-operatively vs. 90 % in KPT group; p = 0.42), similar complication rate (23.5 vs. 33.6 %; p = 0.32) and survival (222 vs. 251 days, p = 0.42). The primary site in the UPT group was confirmed in 10/17 (58.8 %)-all 10 were adenocarcinoma [lung (6) and GI (4)]. DISCUSSION: In our series, the incidence of MSCC due to an unknown primary was 6 %. They had similar overall outcome (neurology post-operatively, complications and survival) to those patients with MSCC from a known primary. Our experience would suggest that we need to treat these patients expeditiously with thorough evaluation and urgent treatment.

Diagnostic evaluation of RT-PCR-ELISA for the detection of rabies virus.

Rabies is primarily a disease of terrestrial and airborne mammals. In most cases, rabies is diagnosed primarily on the basis of clinical symptoms and signs, and a corroborative history of or evidence of an animal bite, death of an animal and incomplete or no vaccination following exposure. The facility for laboratory diagnosis and confirmation of rabies is available in only a few institutions in India. Diagnostic tests using conventional assays like fluorescent antibody test (FAT) are unreliable at times, despite the clinical diagnosis. Currently, there are a number of molecular tests that can be used to complement conventional tests in rabies diagnosis. We have developed and evaluated an RT-PCR-ELISA using a panel of brain tissue samples from rabies suspected animals of various species. This assay was able to detect rabies virus genome in all the 43 samples that were previously tested positive for rabies. Moreover this assay was shown to be 100 % sensitive and specific in detecting the rabies virus genome in post-mortem brain tissue samples from different species of animals. Our pilot study shows the potential of this assay as an alternative diagnostic test when the samples are unsuitable for use in FAT and also a supplementary test to FAT. In addition, the region of nucleoprotein gene amplified using this assay can be used for the molecular investigation of geographical origin of the field strains.

Short duration of lamivudine for the prevention of hepatitis B virus transmission in pregnancy: lack of potency and selection of resistance mutations.

This study sought to assess the antiviral efficacy of lamivudine (LMV) administered during third trimester to reduce maternal viraemia and to identify the emergence of LMV resistance. A prospective observational analysis was performed on 26 mothers with high viral load (>107 IU/mL). Twenty-one women received LMV (treated group) for an average of 53 days (range 22-88 days), and the remaining five formed the untreated control group. Serum samples from two time points were used to measure HBV DNA levels and antiviral drug resistance. The LMV-treated women achieved a median HBV DNA reduction of 2.6-log10 IU/mL. Although end-of-treatment (EOT) HBV DNA in four (18%) LMV-treated women remained at >10(7) IU/mL (± 0.5 log IU/mL), no mother-to-baby transmission was observed. In contrast, a baby from the untreated mother was HBsAg positive at 9 months postpartum. Four technologies were used for drug resistance testing. Only ultra-deep pyrosequencing (UDPS) was sufficiently sensitive to detect minor viral variants down to <1%. UDPS showed that LMV therapy resulted in increased viral quasispecies diversity and positive selection of HBV variants with reverse transcriptase amino acid substitutions at sites associated with primary LMV resistance (rtM204I/V and rtA181T) in four (19%) women. These viral variants were detected mostly at low frequencies (0.63-5.92%) at EOT, but one LMV-treated mother had an rtA181T variant that increased from 2.2% pretherapy to 25.59% at EOT. This mother was also infected with the vaccine escape variant (sG145R), which was inhibited by LMV treatment. LMV therapy during late pregnancy only reduced maternal viraemia moderately, and drug-resistant viral variants emerged.

Raman spectroscopic investigation of the chemopreventive response of naringenin and its nanoparticles in DMBA-induced oral carcinogenesis.

Raman spectroscopy is a vibrational spectroscopic technique that can be used to optically probe the biomolecular changes associated with tumor progression. The aim of the present study is to investigate the biomolecular changes in chemopreventive response of prepared naringenin-loaded nanoparticles (NARNPs) relative to efficacy of free naringenin (NAR) during 7,12-dimethyl benz(a)anthracene (DMBA)-induced oral carcinogenesis by Fourier Transform Raman (FT-Raman) spectroscopy. Oral squamous cell carcinoma (OSCC) was developed in the buccal pouch of golden Syrian hamsters by painting with 0.5% DMBA in liquid paraffin three times a week for 14weeks. Raman spectra differed significantly between the control and tumor tissues, with tumors showing higher percentage signals for nucleic acids, phenylalanine and tryptophan and a lower in the percentage of phospholipids. Moreover, oral administration of free NAR and NARNPs significantly increased phospholipids and decreased the levels of tryptophan, phenylalanine and nucleic acid contents. On a comparative basis, NARNPs was found to have a more potent antitumor effect than free NAR in completely preventing the formation of squamous cell carcinoma and in improving the biochemical status to a normal range in DMBA-induced oral carcinogenesis. The present study further suggest that Raman spectroscopy could be a valuable tool for rapid and sensitive detection of specific biomolecular changes in response to chemopreventive agents.