Sub-chronic cadmium and lead compound exposure induces reproductive toxicity and development of testicular germ cell neoplasia in situ in murine model: Attenuative effects of resveratrol.

Cadmium and lead are widespread, nonbiodegradable heavy metals of perpetual environmental concerns. The present study aimed to evaluate whether sub-chronic exposure to cadmium chloride (CdCl2 ) and lead acetate [Pb(CH3 COO)2 ] induces reproductive toxicity and development of testicular germ cell neoplasia in situ (GCNIS) in swiss albino mice. The effects of resveratrol to reverse the metal-induced toxicity were also analyzed. The mice were randomly divided into four groups for metal treatments and two groups received two different doses of each metal, CdCl2 (0.25 and 0.5 mg/kg) and Pb(CH3 COO)2 (3 and 6 mg/kg). The fourth group received oral doses of 20 mg/kg resveratrol in combination with 0.5 mg/kg CdCl2 or 6 mg/kg Pb(CH3 COO)2 for 16 weeks. Toxic effects of both metals were estimated qualitatively and quantitatively by the alterations in sperm parameters, oxidative stress markers, testicular histology, and protein expressions of the treated mice. Pronounced perturbation of sperm parameters, cellular redox balance were observed with severe distortion of testicular histo-architecture in metal exposed mice. Significant overexpression of Akt cascade and testicular GCNIS marker proteins were recorded in tissues treated with CdCl2 . Notable improvements were observed in all the evaluated parameters of resveratrol cotreated mice groups. Taken together, the findings of this study showed that long-term exposure to Cd and Pb compounds, induced acute reproductive toxicity and initiation of GCNIS development in mice. Conversely, resveratrol consumption abrogated metal-induced perturbation of spermatogenesis, testicular morphology, and the upregulation of Akt cascade proteins along with GCNIS markers, which could have induced the development of testicular cancer.

Lead and cadmium exposure induces male reproductive dysfunction by modulating the expression profiles of apoptotic and survival signal proteins in tea-garden workers.

The aim of this study was to evaluate the impact of Lead (Pb) and Cadmium (Cd) exposure at the molecular level on the reproductive status of tea garden workers in North-East India. Using semen samples, we experimentally determined sperm analysis as well as oxidative stress parameters in all samples and evaluated the expression levels of apoptotic and cell survival proteins [p53, phospho-Akt, nuclear factor-κB (NF-κB, p50 subunit) and B cell lymphoma 2 (Bcl2)]. Our data revealed significant differences in the average heavy metal concentrations and various semen analysis profile between the infertile and normal groups. Increasing Pb and Cd concentrations in semen samples of patients showed positive associations with increasing number of multiple defects in sperm and the level of seminal oxidative stress markers in the high Pb and Cd concentration groups. These groups also exhibited positive correlations between high metal concentrations and the average p53 expression levels, but negative correlations with the mean p-Akt cascade protein levels in sperm cells. In the low Pb and Cd concentrations groups, we also observed reverse mean range and correlation patterns. Therefore, our findings may suggest that graded levels of metal exposure significantly influence the relative fluctuation in the levels of p53 and Akt cascade proteins in the sperm cells of infertile subjects. Furthermore, this may be a regulating factor of sperm cell fate, in turn, determining the fertility outcome of the men working in the tea gardens.

Green Synthesized Ag Nanoparticles for Bio-Sensing and Photocatalytic Applications.

In this work, sensing and photocatalytic activities of green synthesized silver nanoparticles (Ag NPs) are investigated. Ag NPs have been synthesized by the reduction of silver nitrate (AgNO3) using different leaf extracts. An optimum surface plasmon resonance (SPR) behavior is obtained for neem leaf extracts because of the presence of a high concentration of diterpenoids, as evidenced from gas chromatography mass spectroscopy results. The underlying mechanism for the formation of Ag NPs is highlighted. The Ag NPs are in spherical shape and exhibit the hexagonal crystal phase and also show a good stability. The biosensing property of the Ag NPs is evaluated using mancozeb (MCZ) agro-fungicide, and the SPR peak position exhibited a linear response with MCZ concentration. The sensitivity is found to be 39.1 nm/mM. Further, the photocatalytic activity of Ag NPs is tested using 0.5 mM MCZ solution as a model under UV-visible illumination. It is observed that photocatalytic activity is caused by the formation of reactive oxygen species. Therefore, the green synthesized Ag NPs are potential candidates for biosensing and photocatalytic applications.

Successful treatment of refractory lupus erythematosus profundus with the interleukin-6 antagonist tocilizumab: a case report.

Lupus erythematosus profundus (LEP) is an unusual form of cutaneous lupus erythematosus (CLE) characterized by multiple subcutaneous induration and associated with considerable physical and psychological morbidity. The heterogeneity of CLE makes it difficult to understand its underlying pathogenesis and represents a therapeutic challenge. Recently, new insight into the pathogenesis of CLE has implicated various cytokines, opening doors to targeted biologic agents. We report a case of a 23-year-old female who presented with refractory LEP ulcers as an initial presentation of systemic lupus erythematosus. The lesions were resistant to multiple conventional therapies and remarkably responded to tocilizumab.

Fully convolutional multi-scale residual DenseNets for cardiac segmentation and automated cardiac diagnosis using ensemble of classifiers.

Deep fully convolutional neural network (FCN) based architectures have shown great potential in medical image segmentation. However, such architectures usually have millions of parameters and inadequate number of training samples leading to over-fitting and poor generalization. In this paper, we present a novel DenseNet based FCN architecture for cardiac segmentation which is parameter and memory efficient. We propose a novel up-sampling path which incorporates long skip and short-cut connections to overcome the feature map explosion in conventional FCN based architectures. In order to process the input images at multiple scales and view points simultaneously, we propose to incorporate Inception module's parallel structures. We propose a novel dual loss function whose weighting scheme allows to combine advantages of cross-entropy and Dice loss leading to qualitative improvements in segmentation. We demonstrate computational efficacy of incorporating conventional computer vision techniques for region of interest detection in an end-to-end deep learning based segmentation framework. From the segmentation maps we extract clinically relevant cardiac parameters and hand-craft features which reflect the clinical diagnostic analysis and train an ensemble system for cardiac disease classification. We validate our proposed network architecture on three publicly available datasets, namely: (i) Automated Cardiac Diagnosis Challenge (ACDC-2017), (ii) Left Ventricular segmentation challenge (LV-2011), (iii) 2015 Kaggle Data Science Bowl cardiac challenge data. Our approach in ACDC-2017 challenge stood second place for segmentation and first place in automated cardiac disease diagnosis tasks with an accuracy of 100% on a limited testing set (n=50). In the LV-2011 challenge our approach attained 0.74 Jaccard index, which is so far the highest published result in fully automated algorithms. In the Kaggle challenge our approach for LV volume gave a Continuous Ranked Probability Score (CRPS) of 0.0127, which would have placed us tenth in the original challenge. Our approach combined both cardiac segmentation and disease diagnosis into a fully automated framework which is computationally efficient and hence has the potential to be incorporated in computer-aided diagnosis (CAD) tools for clinical application.

Effect of transfer of a poor quality embryo along with a top quality embryo on the outcome during fresh and frozen in vitro fertilization cycles.

OBJECTIVE: To evaluate the impact of a poor quality embryo (PQE) during double ET (DET) with a top quality embryo (TQE) on IVF outcome. DESIGN: A review of prospectively collected data. SETTING: Tertiary level fertility clinic. PATIENT(S): All patients undergoing blastocyst transfers as part of fresh IVF (n = 939) and frozen ET (n = 1,009) cycles performed between 2010 and 2016. INTERVENTION(S): Single ET (SET) with TQE (group 1) was set as control and compared with outcomes for SET with PQE (group 2), DET with 2 TQEs (group 3), PQE plus TQE (group 4), and 2 PQE (group 5). MAIN OUTCOME MEASURE(S): Live births and multiple births. RESULT(S): The live birth rates for group 4 were statistically similar to group 1 during fresh IVF (26.5% vs. 33.7%; odds ratio [OR], 0.95; 95% confidence interval [CI] 0.53-1.7) and frozen ET (24.2% vs. 32.7%; OR, 0.75; 95% CI 0.48-1.2), although there was a trend for lower success. Conversely, multiple births were higher in group 4 for fresh IVF (19% vs. 4.7%; OR, 2.9; 95% CI 1.3-6.6) and frozen ET (10.3% vs. 2.6%; OR, 2.4; 95% CI 1.2-4.9). The live birth rates for group 2 (12.2% for fresh IVF and 14.6% for frozen ET) and group 5 (21.2% for fresh IVF and 14% for frozen ET) were lower and for group 3 were higher (40.8% for fresh IVF and 40.3% for frozen ET) when compared with group 1. Multiple births were significantly higher with DET. CONCLUSION(S): This study does not support DET with one PQE along with a TQE, when there is only one TQE and one or more PQEs available for fresh IVF or frozen ET.

Deep Learning Techniques for Automatic MRI Cardiac Multi-Structures Segmentation and Diagnosis: Is the Problem Solved?

Delineation of the left ventricular cavity, myocardium, and right ventricle from cardiac magnetic resonance images (multi-slice 2-D cine MRI) is a common clinical task to establish diagnosis. The automation of the corresponding tasks has thus been the subject of intense research over the past decades. In this paper, we introduce the "Automatic Cardiac Diagnosis Challenge" dataset (ACDC), the largest publicly available and fully annotated dataset for the purpose of cardiac MRI (CMR) assessment. The dataset contains data from 150 multi-equipments CMRI recordings with reference measurements and classification from two medical experts. The overarching objective of this paper is to measure how far state-of-the-art deep learning methods can go at assessing CMRI, i.e., segmenting the myocardium and the two ventricles as well as classifying pathologies. In the wake of the 2017 MICCAI-ACDC challenge, we report results from deep learning methods provided by nine research groups for the segmentation task and four groups for the classification task. Results show that the best methods faithfully reproduce the expert analysis, leading to a mean value of 0.97 correlation score for the automatic extraction of clinical indices and an accuracy of 0.96 for automatic diagnosis. These results clearly open the door to highly accurate and fully automatic analysis of cardiac CMRI. We also identify scenarios for which deep learning methods are still failing. Both the dataset and detailed results are publicly available online, while the platform will remain open for new submissions.

Mucocele: A diagnostic dilemma!!

Oral mucocele is the most common benign lesion of minor salivary gland caused due to any form of mechanical trauma to the excretory duct of the gland. It is of two types - mucous extravasation phenomenon and mucous retention type. Extravasation type is more common. Here, we report a case of a 65-year-old male patient with a complaint of painful swelling in the lower lip for 1 month with a history of trauma in the lower lip. It was clinically diagnosed as traumatic fibroma, and an excisional biopsy was done. However, the histopathology was basically that of mucous extravasation phenomenon coexisting with the features of a reactive lesion implicating the potential role of chronic irritation in the impairment of normal healing process. Considering the fact that long-standing chronic inflammation in a benign lesion can even lead to malignant transformation of that lesion; here, we highlight the role of a good clinicopathologic correlation and the significance of prompt intervention and treatment. Furthermore, emphasizing the potential need for postoperative follow-up by the dentist, how much ever trivial the lesion may appear to be in the best interest of the patient.

Multidisciplinary management of hypohydrotic ectodermal dysplasia - a case report.

Hypohydrotic ectodermal dysplasia is a hereditary disorder, which affects ectodermal derivatives. It manifests several abnormalities of the teeth, and is commonly inherited through female carriers. This case report presents a patient with compromised esthetics and function. A multidisciplinary approach was planned involving an oral pathologist, endodontist, orthodontist and a prosthodontist.

Oral mucocutaneous lesions - a comparative clinicopathological and immunofluorescence study.

BACKGROUND: Oral mucosa is often affected by many diseases including mucocutaneous disorders. The diagnoses of these disorders are primarily based on history, clinical features, and histopathology. For the past few years' immunofluorescence techniques emerged as an important tool to study the pathogenesis and in the diagnosis of oral mucocutaneous and vesiculobullous disorders. The present study was designed to carry out retrospective and prospective analysis of oral mucocutaneous lesions to elucidate the clinicopathologic features and its immunofluorescence findings. MATERIALS AND METHODS: A total of 70 subjects with oral mucocutaneous lesions were retrieved from the oral pathology files of Tamil Nadu Govt. Dental College and their clinical features were evaluated, and the histopathology was also evaluated with the help of hematoxylin and eosin stained sections. For the prospective study, biopsy from 12 subjects with oral mucocutaneous lesions was subjected to routine histopathological examination and DIF to evaluate the consistency of the methods. RESULTS: In the retrospective analysis of 70 subjects with oral mucocutaneous lesions in relation to clinical features and histopathology, most of the findings were similar to the previous studies except for few criteria like male predilection in lichen planus and mucous membrane pemphigoid (MMP) and more prevalence of pemphigus vulgaris than MMP (2:1). In the prospective analysis of 12 subjects, the histopathological diagnosis was consistent with DIF study in 66% of cases. CONCLUSION: The diagnostic efficiency of oral mucocutaneous lesions can be improved by modern tools like DIF studies in addition to traditional methods like clinical and histopathology.

OMICS: Current and future perspectives in reproductive medicine and technology.

Many couples present fertility problems at their reproductive age, and although in the last years, the efficiency of assisted reproduction techniques has increased, these are still far from being 100% effective. A key issue in this field is the proper assessment of germ cells, embryos and endometrium quality, in order to determine the actual likelihood to succeed. Currently available analysis is mainly based on morphological features of oocytes, sperm and embryos and although these strategies have improved the results, there is an urgent need of new diagnostic and therapeutic tools. The emergence of the - OMICS technologies (epigenomics, genomics, transcriptomics, proteomics and metabolomics) permitted the improvement on the knowledge in this field, by providing with a huge amount of information regarding the biological processes involved in reproductive success, thereby getting a broader view of complex biological systems with a relatively low cost and effort.

Effective control of polymyalgia rheumatica with tocilizumab.

Despite their disadvantages, glucocorticoids (GCs) remain a mainstay of therapy for polymyalgia rheumatica (PMR). Second-line antirheumatic and immune-modulatory drugs are not infrequently required because of disease relapses during GC tapering and GC adverse effects. Therapy with methotrexate or with an anti-tumor necrosis factor drug showed modest efficacy in this situation. Tocilizumab (TCZ) is an anti-interleukin 6 receptor antibody that is being recently studied in the treatment of PMR patients who are intolerant or refractory to GCs, especially after failure of a second-line agent. We report a case of PMR in which GCs were stopped because of adverse effects despite good response. The condition responded to neither methotrexate nor etanercept. Treatment with TCZ has led to significant improvement of the patient's clinical and biochemical PMR activity parameters, and she was kept in a solid remission for 1 year without any TCZ-related adverse effects. Tocilizumab is a promising drug in the management of PMR. Further studies are required to clearly define the indications and duration of TCZ therapy in the management of PMR.

Laboratory handling of epididymal and testicular spermatozoa: What can be done to improve sperm injections outcome.

Spermatozoa from azoospermic males can be retrieved from either the epididymis or the testis, depending on the type of azoospermia, using different surgical methods such as percutaneous epididymal sperm aspiration (PESA), testicular sperm aspiration (TESA), testicular sperm extraction (TESE), and microsurgical testicular sperm extraction (micro- TESE). After collecting the epididymal fluid or testicular tissue, laboratory techniques are used to remove contaminants, cellular debris, noxious microorganisms, and red blood cells. Processed spermatozoa may be used for intracytoplasmic sperm injection or eventually be cryopreserved. However, spermatozoa collected from either the epididymis or the testis are often compromised and more fragile than ejaculated ones. Therefore, sperm processing techniques should be used with great caution to avoid jeopardizing the sperm fertilizing potential in treatment cycles. In this review, we describe the current methods for processing surgically-retrieved specimens, either fresh or frozen- thawed, and provide the tips and pitfalls for facilitating the handling of such specimens. In addition, we present the available laboratory tools to aid in the identification of viable immotile spermatozoa to be used in conjunction with assisted reproductive techniques. Review of the literature was carried out using PubMed and Science Direct search engines.

Oocyte developmental competence and embryo development: impact of lifestyle and environmental risk factors.

Oocyte development is the end result of a sophisticated biological process that is hormonally regulated and produced by highly specialized cellular lines that differentiate in early embryo/fetal development. Embryo development is initially regulated by maternal transcripts until replaced by embryonic genomic expression. Then, an assortment of hormones and local environmental factors in various concentrations along the reproductive tract (e.g. fallopian tube, endometrial lining) provide the protection, nutrients and means of communication for the embryo to implant and develop. Both oocytes and embryos are susceptible to environmental, occupational and lifestyle exposures that can exert direct toxic effects and disrupt hormones. While some exposures may produce reversible changes, others, especially those damaging germinal cells in utero or during prepuberty, may result in permanent sequelae that continue in future generations. This article reviews the main factors that affect female fertility and their possible influence on human reproduction. Some lifestyles, xeno-oestrogens and heavy metals are already known to compromise female reproductive function. Nonetheless, many questions remain and little is known about the effect of many other factors on female fertility.

The genetic causes of male factor infertility: a review.

OBJECTIVE: To illustrate the necessity for an enhanced understanding of the genetic basis of male factor infertility, to present a comprehensive synopsis of these genetic elements, and to review techniques being utilized to produce new insights in fertility research. BACKGROUND: Male factor infertility is a complex disorder that affects a large sector of the population; however, many of its etiologies are unknown. By elucidating the underlying genetic basis of infertile phenotypes, it may be possible to discover the causes of infertility and determine effective treatments for patients. METHOD(S): The PubMed database was consulted for the most relevant papers published in the last 3 years pertaining to male factor infertility using the keywords "genetics" and "male infertility." RESULT(S): Advances have been made in the characterization of the roles of specific genes, but further research is necessary before these results can be used as guidelines for diagnosing and treating male factor infertility. The accurate transmission of epigenetic information also has considerable influence on fertility in males and on the fertility of their offspring. CONCLUSION(S): Analysis of the genetic factors that impact male factor infertility will provide valuable insights into the creation of targeted treatments for patients and the determination of the causes of idiopathic infertility. Novel technologies that analyze the influence of genetics from a global perspective may lead to further developments in the understanding of the etiology of male factor infertility through the identification of specific infertile phenotype signatures.

The in vitro effect of benzo[a]pyrene on human sperm hyperactivation and acrosome reaction.

OBJECTIVE: To evaluate the in vitro effect of benzo[a]pyrene on sperm hyperactivation and acrosome status in normozoospermic semen samples of nonsmokers analyzed by computer-assisted semen analysis (CASA). DESIGN: Experimental in vitro study. SETTING: Andrology laboratory. PATIENT(S): Thirteen proven fertile, normozoospermic, and nonsmoking men. INTERVENTION(S): Spermatozoa were washed free of seminal plasma and were treated with different concentrations of benzo[a]pyrene and compared with controls treated with medium alone. The benzo[a]pyrene concentrations were: 100, 50, 25, and 12.5 microg/mL. MAIN OUTCOME MEASURE(S): Effect of varying concentrations of benzo[a]pyrene on sperm hyperactivation and acrosomal reaction. RESULT(S): A statistically significant increase in sperm hyperactivation was observed in presence of benzo[a]pyrene at concentrations of >or=50 microg/mL. The result of the acrosome halo test showed that concentrations of benzo[a]pyrene >or=25 microg/mL statistically significantly decreased the percentage of halo formation, indicating an inappropriate (false) acrosome reaction. CONCLUSION(S): Benzo[a]pyrene statistically significantly affected sperm functional competence as evidenced by increased hyperactivation as well as premature acrosomal reaction.

Semen quality and age-specific changes: a study between two decades on 3,729 male partners of couples with normal sperm count and attending an andrology laboratory for infertility-related problems in an Indian city.

OBJECTIVE: To compare the semen quality and age-specific changes in men between the 1980s and 2000s. DESIGN: Prospective study. SETTING: Andrology laboratory, University of Calcutta, India. PATIENT(S): A semen sample was obtained from 3729 men presenting for infertility problems in two distinct decades, that is, between 1981-85 and 2000-2006. INTERVENTION(S): Subjects with sperm count >20 x 10(6)/mL without any extreme pathological disorders were selected. Samples having a major liquefaction problem were excluded. MAIN OUTCOME MEASURE(S): A standard World Health Organization procedure for semen analysis was performed that included assessment of volume, sperm concentration, and percentage motility. The motility parameters were further classified into forward progressive motility and nonprogressive motility. RESULT(S): The present large-scale study confirms a significant decline in the sperm motility parameters and seminal volume in the present decade. However, no change in overall sperm concentration was noted. A decline was seen in sperm motility with increasing age in both decades. CONCLUSION(S): There are significant changes in sperm motility and volume between the two decades, and the age-related changes in semen parameters are also different in the two decades.

Expression of metallothionein-1 (MT-1) mRNA in the rat testes and liver after cadmium injection.

Metallothioneins (MTs) belong to the family of stress proteins that are present in the majority of living organisms. The MTs play an important task in detoxifying heavy metals. The mammalian scrotal testis is known to be susceptible to cadmium (Cd) exposure. The present work focuses on the MT-1 isoform and aims to ascertain and confirm previous findings to answer whether rodent testes indeed contain MT-1 mRNA, whether its level is increased with Cd injection in liver and testes, and lastly what is the relative difference in the expression of MT-1 mRNA in liver and testes both with and without Cd injection. Adult male Wistar rats weighing 270-290 g received a subcutaneous injection of 4.0 mumol Cd/kg and were sacrificed by cervical dislocation 6 h later. RNA was isolated from testes as well as the liver. There were 2 replicates per treatment for RNA analyses. MT-1 mRNA levels were determined by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis and then assessed by densitometry scanning. The results of RT-PCR clearly demonstrated that the rodent testes express MT-1 mRNA. The densitometry data shows that the expression of MT-1 mRNA increases with Cd treatment in testes. The relative level of MT1-mRNA is greater in the control-liver than in the control-testes. However, upon Cd injection, the level of testes MT-1 mRNA increases 2.16 fold. These results suggest that the testes respond to Cd for at least 6 h post injection through a transcriptional mechanism.

Markers of oxidative stress and sperm chromatin integrity.

Oxidative stress (OS) is an imbalance between the amount of reactive oxygen species (ROS) produced and the ability of the antioxidants to scavenge these. OS has been established as a major etiological cause of male infertility. High levels of ROS are harmful and cause damage to sperm nuclear DNA. Evaluation of OS-related damage to spermatozoa is therefore highly relevant in assisted reproductive techniques (ART) such as intracytoplasmic sperm injection (ICSI). ICSI is an effective therapy for severe male factor infertility that bypasses the majority of reproductive tract deficiencies. Despite the controversial findings in the existing literature, there is now enough evidence to show that sperm DNA damage is detrimental to reproductive outcomes. In addition to impairment of fertility, such damage might increase the transmission of genetic diseases to the offspring. Standardization of protocols to assess ROS, antioxidant status, and DNA damage is very important for implementation of these tests in clinical practice. Estimation of seminal ROS levels and extent of sperm DNA damage, especially in an infertile male, may help develop new therapeutic strategies and improve the success of ART.