Nimbolide Inhibits SOD2 to Control Pancreatic Ductal Adenocarcinoma Growth and Metastasis.

Reactive oxygen species are frequently associated with various cancers including pancreatic ductal adenocarcinomas (PDACs). Superoxide dismutase 2 (SOD2) is an enzyme that plays an important role in reactive oxygen species (ROS) signaling. Investigating the molecular function and biological functions of SOD2 can help us develop new therapeutic options and uncover new biomarkers for PDAC diagnosis and prognosis. Here, we show that nimbolide (NB), a triterpene limonoid, effectively blocks the growth and metastasis of PDACs by suppressing the expression and activity of SOD2. To identify the role of SOD2 in NB-induced anticancer activity, we used RNA interference to silence and plasmid transfection to overexpress it. Silencing SOD2 significantly reduced the growth and metastatic characteristics like epithelial-to-mesenchymal transition, invasion, migration, and colony-forming capabilities of PDACs, and NB treatment further reduced these characteristics. Conversely, the overexpression of SOD2 enhanced these metastatic characteristics. ROS signaling has a strong feedback mechanism with the PI3K/Akt signaling pathway, which could be mediated through SOD2. Finally, NB treatment to SOD2-overexpressing PDAC xenografts resulted in significant inhibition of tumor growth and metastasis. Overall, this work suggests that NB, a natural and safe phytochemical that silences SOD2 to induce high levels of ROS generation, results in increased apoptosis and reduced growth and progression of PDACs. The role of SOD2 in regulating NB-induced ROS generation presents itself as a therapeutic option for PDACs.

Perinatal Outcomes In Monozygotic Pregnancies Resulting From Assisted Reproductive Technology Procedures: A Single-Center 6-Year Experience Based On A Large Cohort Of Pregnancies.

OBJECTIVE: Monozygotic twin (MZT) pregnancies increase the risk of maternal and infant mortality and include many complications. The present study describes our assisted reproductive technology (ART) procedures from the viewpoint of perinatal outcomes in MZT pregnancies. METHODS: In this retrospective clinical cross-sectional study, 1159 in vitro fertilization (IVF) cycles performed between October 2014 and December 2019 were reviewed and perinatal outcomes and general clinical conditions analyzed. RESULTS: Sixteen MZT pregnancies were observed, resulting in an incidence of 1.38%. The MZT pregnancy incidence for patients aged ≤35 and >35 years were 0.2% and 1.1%, respectively. Eight MZT pregnancies resulted in live births, while five ended in miscarriage. A significant positive correlation was found between the number of attempts and the age of female (r:0.674; p=0.004) and male (r:0.657; p=0.006) partners. Cumulus-Oocytes Complexes (COC) (r:0.635; p=0.008), Metaphase II Oocyte (MIIO) (r:0.627; p=0.009), Pronucleus Oocyte (PO) (r:0.585; p=0.017) were correlated with serum AMH levels. The number of MZT was positively correlated with male partner age (r:0.527; p=0.036) and negatively correlated with embryo transfer day (ETd) (r:-0.548; p=0.028). CONCLUSIONS: The incidence of MZT pregnancies observed in this study was similar to the incidence reported in the literature, although risk was more pronounced among women aged >35 years. Due to potential risks for mothers and fetuses, MZT pregnancies may become a problem as the number of individuals seeking IVF continues to increase.

Transportation as a Disease Vector in COVID-19: Border Mobility and Disease Spread.

More than a year after COVID-19 was declared a pandemic by the World Health Organization, the U.S.A. and Mexico rank first and fourth, respectively, with regard to the number of deaths. From March 2020, nonessential travelers were not allowed to cross the border into the U.S.A. from Mexico via international land ports of entry, which resulted in a more than 50% decrease in the number of people crossing the border. However, border communities still face a higher number of cases and faster community spread compared with those without international land ports of entry. This paper established an econometric model to understand the effects of cross-border mobility and other socioeconomic parameters on the speed of spread. The model was developed at the U.S. county level using data from all 3,141 counties in the U.S.A. Additionally, a follow-up U.S. county comparative analysis was developed to examine the significance of having a border crossing between the U.S.A. and Mexico for U.S. counties. The findings of the analysis revealed that the variables having a significant effect are as follows: population density; number of people per household; population in the 15-65 age group; median household income; mask use; number of visits to transit stations; number of visits to workplace; overall mobility; and having a border crossing to Mexico within county limits. The comparative analysis found that U.S. counties with border crossings have an average of 123 cases per 1,000 population whereas their counterparts without border crossings only have 90 cases per 1,000 population.

Analysis of the Prevalence and Quantification of viral load of different human papillomavirus types in Turkish women population.

BACKGROUND: Persistent infection with high-risk (HR) Human Papilloma Virus (HPV) genotypes has been shown to play a significant role in the development of cervical intraepithelial neoplasia (CIN) and CC (cervical cancer). The present study aimed to determine the distribution and quantification of viral load of HPV genotypes in numerous genital samples obtained from women undergoing routine gynaecological care in different regions of Turkey. METHODS: HPV typing was done by HPV QUANT-21 Quantitative RT-PCR Kit®, which is intended for the specific identification and quantification of low-risk (HPV 6, 11, 44) and high-risk (HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82) from female subjects in Turkey. RESULTS: From the total of 5975 samples, 2777 were positive for at least one HPV genotype, with an overall frequency of 46.4%. 1695 patients were positive for only one or more LR-HPV genotypes (61%) and 812 patients were positive for one or more HR-HPV genotypes (29%). The frequency of LR-HPV genotypes was 31.4%, while the frequency of HR-HPV genotypes was 118.8%. Our tecnology had a positive advantage to calculate the concentration of each genotypes. Although genotype 52 ranked fifth in frequency, it showed the highest mean concentration, with a value of 5.38 log (copies/sample). CONCLUSION: The presence and genotype of viruses before HPV vaccination have also gained importance. The data obtained would provide guidance for prevention strategies, mainly of vaccination. We decided to add a new estimate to the effectiveness of currently available HPV vaccines and the development of screening programs to prevent and decrease the incidence of CC in Turkey. Further studies would be planned to measure and define the high infection level that can lead to the development of cervical neoplasia. Using this tecnology could give us a clinical desicion to degree the cytological changes.

Comparison of complication rates after transvaginal ultrasound-guided oocyte pick-up procedures with respect to ovarian response.

OBJECTIVE: The aim of this study was to compare the complication rates of oocyte pick-up (OPU) procedures via transvaginal ultrasonography in patients with different levels of ovarian reserve. METHODS: In total, 789 patients who underwent OPU procedures for in vitro fertilization (IVF) were included in the study. RESULTS: Individuals with normal ovarian reserve had a 2.947-fold higher risk of complications in OPU procedures than individuals with low ovarian reserve, and individuals with high ovarian reserve had a 7.448-fold higher risk of complications than individuals with low ovarian reserve. In addition, a higher number of IVF trials was associated with an increased risk of complications. CONCLUSION: The results of this study show that OPU has a higher risk of complications, particularly severe pain, in patients with high ovarian reserve. It is thought that complications can be reduced by preferring mild stimulation in patients with high ovarian reserve. Collecting fewer oocytes is also associated with a lower risk of complications from OPU. Even if a patient's reserve is very good, fewer and higher-quality oocytes should be targeted with the use of the lowest possible dose of drugs.

Association of increased levels of omentin-1 and carotid intima-media thickness with early signs of cardiovascular risk in patients with polycystic ovary syndrome: A prospective case control study.

AIM: Omentin-1, an adipokine primarily secreted from visceral adipose tissue, regulates metabolic homeostasis, whereas carotid intima-media thickness (CIMT) is a valid marker for early diagnosis of atherosclerosis. We aimed to investigate whether polycystic ovary syndrome (PCOS) is related to levels of omentin-1 and CIMT and demonstrate the association with cardiovascular risk. METHODS: A prospective case-control study was performed on 47 PCOS patients and 35 healthy subjects. Hormonal and metabolic parameters, levels of serum omentin-1, and CIMT were evaluated in the two groups. RESULTS: The right, left, total CIMT, and levels of serum omentin-1 were significantly higher in the PCOS group than in the healthy controls (p = 0.015, p = 0.009, p = 0.021, and p = 0.002, respectively). Using multiple linear and backward elimination regression analyses, serum omentin-1 was found to be independently associated with age and Ln-transformed homeostasis model assessment of insulin resistance, and cardiovascular parameters were independently associated with body mass index (BMI). In the PCOS group, the means of right, left, and total CIMT were significantly higher in the subgroup with BMI levels ≥25 kg/m2 compared to the subgroup with BMI levels <25 kg/m2 (p < 0.001 for all comparisons). CONCLUSION: A significant increase in CIMT is associated with cardiovascular risk in patients with PCOS. Levels of serum omentin-1 were found to be significantly higher in early PCOS cases at a younger age, acting as a protective acute-phase reactant.

Phytochemicals as an Alternative or Integrative Option, in Conjunction with Conventional Treatments for Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is the most abundant form of liver cancer. It accounts for 75-85% of liver cancer cases and, though it ranks globally as the sixth most common cancer, it ranks second in cancer-related mortality. Deaths from HCC are usually due to metastatic spread of the cancer. Unfortunately, there are many challenges and limitations with the latest HCC therapies and medications, making it difficult for patients to receive life-prolonging care. As there is clearly a high demand for alternative therapy options for HCC, it is prudent to turn to plants for the solution, as their phytochemicals have long been used and revered for their many medicinal purposes. This review explores the promising phytochemical compounds identified from pre-clinical and clinical trials being used either independently or in conjunction with already existing cancer therapy treatments. The phytochemicals discussed in this review were classified into several categories: lipids, polyphenols, alkaloids, polysaccharides, whole extracts, and phytochemical combinations. Almost 80% of the compounds failed to progress into clinical studies due to lack of information regarding the toxicity to normal cells and bioavailability. Although large obstacles remain, phytochemicals can be used either as an alternative or integrative therapy in conjunction with existing HCC chemotherapies. In conclusion, phytochemicals have great potential as treatment options for hepatocellular carcinoma.

Investigating Fetuin-A and Paraoxonase-1 Activity as Markers in Polycystic Ovary Syndrome Based on Body Mass Index: A Prospective Case-Control Study.

Introduction Polycystic ovary syndrome (PCOS) may have an increased risk for the development of systemic and metabolic pathogenesis such as cardiovascular diseases, insulin resistance (IR), diabetes mellitus (DM) and dyslipidemia. However, there is no reliable marker to show the relation. Fetuin-A is an adipokine whereas paraoxonase-1 (PON-1) is a high-density lipoprotein-linked enzyme to demonstrate oxidative stress. This study aimed to evaluate serum fetuin-A and PON-1 levels in infertile PCOS women based on body mass index (BMI). Methods A prospective case-control study in a university setting was designed. A total of 88 patients admitted to the Giresun University Faculty of Medicine Gynecology Clinic between February and April 2021 were included in the study. The subjects were divided as follows: PCOS-low-BMI (BMI≤25) vs. Controls-low-BMI (BMI≤25) and PCOS-high-BMI (BMI>25) vs. Controls-high-BMI (BMI>25). Those who had at least two criteria of the 2003 Rotterdam Consensus were diagnosed with PCOS. Serum fetuin-A and PON-1 levels were compared. Results The mean levels of fetuin-A were not significantly different in the groups (p=0.955). Serum PON-1 levels were lower in the PCOS group (109.1±61.4 vs. 140.1±80.0; p=0.040), but it lost significance with adjusted values for covariants as age and BMI. Although PON-1 was not significantly different in the PCOS group of BMI<25 kg/m2 subgroup, it was significantly lower in the PCOS group of BMI≥25 kg/m2 subgroup (p=0.820 vs. p=0.048). Conclusion Serum fetuin-A activity did not differ with PCOS. Serum PON-1 might be a promising and research-worthy marker, especially for obese PCOS patients.

miRNome and Functional Network Analysis of PGRMC1 Regulated miRNA Target Genes Identify Pathways and Biological Functions Associated With Triple Negative Breast Cancer.

BACKGROUND: Increased expression of the progesterone receptor membrane component 1, a heme and progesterone binding protein, is frequently found in triple negative breast cancer tissue. The basis for the expression of PGRMC1 and its regulation on cellular signaling mechanisms remain largely unknown. Therefore, we aim to study microRNAs that target selective genes and mechanisms that are regulated by PGRMC1 in TNBCs. METHODS: To identify altered miRNAs, whole human miRNome profiling was performed following AG-205 treatment and PGRMC1 silencing. Network analysis identified miRNA target genes while KEGG, REACTOME and Gene ontology were used to explore altered signaling pathways, biological processes, and molecular functions. RESULTS: KEGG term pathway analysis revealed that upregulated miRNAs target specific genes that are involved in signaling pathways that play a major role in carcinogenesis. While multiple downregulated miRNAs are known oncogenes and have been previously demonstrated to be overexpressed in a variety of cancers. Overlapping miRNA target genes associated with KEGG term pathways were identified and overexpression/amplification of these genes was observed in invasive breast carcinoma tissue from TCGA. Further, the top two genes (CCND1 and YWHAZ) which are highly genetically altered are also associated with poorer overall survival. CONCLUSIONS: Thus, our data demonstrates that therapeutic targeting of PGRMC1 in aggressive breast cancers leads to the activation of miRNAs that target overexpressed genes and deactivation of miRNAs that have oncogenic potential.

Prolonged luteal phase support with progesterone may increase papules and plaques of pregnancy frequency in pregnancies through in vitro fertilization

Abstract Background: Pruritic urticarial papules and plaques of pregnancy development may have a strong relationship with hormone treatments during in vitro fertilization and hormonal changes during pregnancy. Objectives: The aim of this study was to evaluate and compare the frequency of papules and plaques of pregnancy and related factors in in vitro fertilization pregnancies and spontaneous pregnancies. Methods: In this study, 517 in vitro fertilization pregnancies and 1253 spontaneous pregnancies were retrospectively reviewed for papules and plaques of pregnancy frequency. The diagnosis of papules and plaques of pregnancy was performed by referral to the dermatology department and according to the typical clinical manifestations of the disease. Results: The papules and plaques of pregnancy was more common in all in vitro fertilization pregnancies (including single pregnancies) than in spontaneous pregnancies. Age, Rh positivity, mother weight gain, onset of disease during gestation, duration of disease, birth weight and the frequency of male fetus were similar between the two groups (p > 0.05). The rate of multiple pregnancies was higher in in vitro fertilization pregnancies with papules and plaques of pregnancy than in vitro fertilization pregnancies without papules and plaques of pregnancy (p < 0.001). Duration of progesterone treatment was also significantly longer in in vitro fertilization pregnancies with papules and plaques of pregnancy compared to in vitro fertilization pregnancies without papules and plaques of pregnancy (p < 0.001). Study limitations: The limitations of the study were the retrospective and single-centered design. Conclusion: The results of this study indicate that increased progesterone dosage or prolonged treatment may play a role in the pathogenesis papules and plaques of pregnancy.

Spectroscopy analysis of endometrial metabolites is a powerful predictor of success of embryo transfer in women with implantation failure: a preliminary study.

OBJECTIVE: To investigate whether prior endometrium spectroscopy predicts the success of embryo transfer in patients with recurrent implantation failure (RIF). MATERIALS AND METHODS: Twenty women with RIF who were scheduled for IVF-ET and six fertile women were included the study. All women with RIF and fertile controls underwent endometrium spectroscopy before embryo transfer. A multi-voxel spectroscopy sequence was used for the detection of choline (Cho), creatine (Cr), lactate, and lipids. Women with RIF were divided into two subgroups as successful (n = 8) and unsuccessful RIF (n = 12) according to both Cho and Cr metabolite characteristics and whether pregnancy was achieved. Associations between the metabolite intensities of the RIF subjects and the implantation rate (IR) and clinical pregnancy rate (CPR) were assessed. RESULTS: While 8 of 20 RIF cases became pregnant, pregnancy could not be achieved in 12 cases. The common spectroscopy finding in all 8 cases who conceived was high Cho and low lactate. The main metabolite change detected in 12 patients who could not conceive was the increase in lactate and lipid signals. The cutoff value of Cho, Cr, lactate and lipid were 1.01 ppm, 1.44 ppm, 0.86 ppm and 1.22 ppm respectively in patients who achieved pregnancy following ET. A positive and significant correlation was found between Cho and Cr intensities and IR or CPR. CONCLUSIONS: Receptive endometrium represents some specific metabolites in spectroscopy that can be used for prediction of the success or failure of embryo transfer in women suffering implantation failure.

Prolonged luteal phase support with progesterone may increase papules and plaques of pregnancy frequency in pregnancies through in vitro fertilization.

BACKGROUND: Pruritic urticarial papules and plaques of pregnancy development may have a strong relationship with hormone treatments during in vitro fertilization and hormonal changes during pregnancy. OBJECTIVES: The aim of this study was to evaluate and compare the frequency of papules and plaques of pregnancy and related factors in in vitro fertilization pregnancies and spontaneous pregnancies. METHODS: In this study, 517 in vitro fertilization pregnancies and 1253 spontaneous pregnancies were retrospectively reviewed for papules and plaques of pregnancy frequency. The diagnosis of papules and plaques of pregnancy was performed by referral to the dermatology department and according to the typical clinical manifestations of the disease. RESULTS: The papules and plaques of pregnancy was more common in all in vitro fertilization pregnancies (including single pregnancies) than in spontaneous pregnancies. Age, Rh positivity, mother weight gain, onset of disease during gestation, duration of disease, birth weight and the frequency of male fetus were similar between the two groups (p>0.05). The rate of multiple pregnancies was higher in in vitro fertilization pregnancies with papules and plaques of pregnancy than in vitro fertilization pregnancies without papules and plaques of pregnancy (p<0.001). Duration of progesterone treatment was also significantly longer in in vitro fertilization pregnancies with papules and plaques of pregnancy compared to in vitro fertilization pregnancies without papules and plaques of pregnancy (p<0.001). STUDY LIMITATIONS: The limitations of the study were the retrospective and single-centered design. CONCLUSION: The results of this study indicate that increased progesterone dosage or prolonged treatment may play a role in the pathogenesis papules and plaques of pregnancy.

Enhancement of Zika virus infection by antibodies from West Nile virus seropositive individuals with no history of clinical infection.

BACKGROUND: Recent outbreaks of Zika Virus (ZIKV) infection and associated microcephaly has raised multiple scientific questions. The close antigenic relatedness between flaviviruses makes diagnosis of specific infection difficult. This relatedness also raises the potential of Antibody Dependent Enhancement (ADE) via cross reactive antibodies to flaviviruses like West Nile Virus (WNV) and Dengue Virus (DENV). Asymptomatic WNV infections are endemic throughout the US creating a large proportion of the population that is seropositive for WNV antibodies. Whether these sero-positive individuals potentially carry ZIKV enhancing antibodies remains unknown. RESULTS: Serum samples obtained from human subjects with symptomatic or asymptomatic WNV infection from a WNV endemic region in Texas were tested for their ability to enhance or neutralize ZIKV infection. Sero-surveillance data demonstrated a ~ 7% prevalence for WNV antibodies in the population. Sera from both symptomatic and asymptomatic WNV seropositive donors effectively neutralized WNV and to some extent DENV infection. Interestingly, WNV+ sera failed to inhibit ZIKV while significantly enhancing infection. Conversely, ZIKV specific sera effectively neutralized ZIKV, with ADE only evident at lower concentrations. The enhancement of ZIKV via WNV antibody positive sera was likely due to non-neutralizing Envelope (E) antibodies as seen with monoclonal ZIKV E antibodies. CONCLUSIONS: Overall, our findings suggest that WNV antibodies in the sera significantly enhance ZIKV infection in Fc receptor positive cells with limited neutralization activity. Further studies in more relevant models of ADE will be needed to confirm the relevance of these findings in vivo.

Combination gene therapy for HIV using a conditional suicidal gene with CCR5 knockout.

BACKGROUND: Gene therapy approaches using hematopoietic stem cells to generate an HIV resistant immune system have been shown to be successful. The deletion of HIV co-receptor CCR5 remains a viable strategy although co-receptor switching to CXCR4 remains a major pitfall. To overcome this, we designed a dual gene therapy strategy that incorporates a conditional suicide gene and CCR5 knockout (KO) to overcome the limitations of CCR5 KO alone. METHODS: A two-vector system was designed that included an integrating lentiviral vector that expresses a HIV Tat dependent Thymidine Kinase mutant SR39 (TK-SR39) and GFP reporter gene. The second non-integrating lentiviral (NIL) vector expresses a CCR5gRNA-CRISPR/Cas9 cassette and HIV Tat protein. RESULTS: Transduction of cells sequentially with the integrating followed by the NIL vector allows for insertion of the conditional suicide gene, KO of CCR5 and transient expression of GFP to enrich the modified cells. We used this strategy to modify TZM cells and generate a cell line that was resistant to CCR5 tropic viruses while permitting infection of CXCR4 tropic viruses which could be controlled via treatment with Ganciclovir. CONCLUSIONS: Our study demonstrates proof of principle that a combination gene therapy for HIV is a viable strategy and can overcome the limitation of editing CCR5 gene alone.

The utility of detecting ovulation to predict success in ovulation induction and intrauterine insemination cycles - a prospective observational study.

OBJECTIVES: The success of ovulation induction-intrauterine insemination (OI-IUI) procedures may be limited by the absence of ovulation detection. The aim of this study was to evaluate the empirical use of ultrasonography and luteal phase progesterone (P4) as ovulation indicators and determine its effect on pregnancy outcome in OI-IUI cycles. MATERIAL AND METHODS: This prospective observational study, which was performed in a university setting, included 107 women with unexplained infertility. Following OI, IUI was performed 36 hours after human chorionic gonadotropin (hCG). P4 was measured 72-96 hours after hCG. At the same time, the appearance of ovaries and signs indicative of ovulation, which are decreased follicle dimensions, irregularity of follicular walls, and the presence of free fluid in the Douglas pouch, were noted. RESULTS: In 58 patients (54.2%), ovulation was detected at the P4 level of > 10 ng/mL. Eighty-nine patients had ultrasound images suggestive of ovulation. However, only 50 of these were confirmed ovulation as indicated using P4. Implantation was observed in a total of 13 patients (12.1%). All patients were in the ovulation detected group with P4 > 10 ng/mL (AUC: 0.750; p = 0.004). P4 of > 21.5 ng/mL detected successful ovulation and was strongly associated with implantation with 77% sensitivity and 61% specificity (OR: 9.9; 95% CI: 2.4-41.2). Body mass index (BMI) > 23.9 kg/m2 was a reliable anovulation indicator as a secondary outcome (AUC: 0.696; p = 0.02). CONCLUSIONS: In 45.8% of the patients, ovulation did not occur even with OI treatment. The association of progesterone measurement and ovarian ultrasound scanning between 72 and 96 hours after hCG treatment can be used to detect ovulation. In doing so, we can find the optimal treatment for patients with infertility in their next cycle.

Comparison of patients undergoing vaginal hysterectomy with sacrospinous ligament fixation, laparoscopic hysterectomy with sacrocolpopexy and abdominal hysterectomy with sacrocolpopexy in terms of postoperative quality of life and sexual function.

AIM: To investigate whether there is a significant difference among the patients who underwent vaginal hysterectomy with sacrospinous ligament fixation (VH + SSLF), laparoscopic hysterectomy with sacrocolpopexy (LH + SCP) and abdominal hysterectomy with sacrocolpopexy (AH + SCP) in terms of sexual function and quality of life. MATERIAL AND METHOD: Sixty-five patients undergoing vaginal hysterectomy with sacrospinous ligament fixation (VH + SSLF), laparoscopic hysterectomy with sacrocolpopexy (LH + SCP) or abdominal hysterectomy with sacrocolpopexy (AH + SCP) participated in the study. The Quality of Life Scale and Sexual Function Scale Index (PISQ-12) were used to see whether there is a significant difference among the three groups (VH + SSLF, LH + SCP, AH + SCP) at least 1 year after surgery. The Pelvic Floor Distress Inventory-20 (PFDI-20) Scale consisting of Pelvic Organ Prolapse Distress Inventory (POPDI-6), Urinary Distress Inventory (UDI-6), and Colorectal-Anal Distress Inventory (CRADI-8) was used to evaluate the functional outcomes. RESULTS: The participants had a mean age of 60 ± 8.79 years. The mean PFDI-20 score in the VH + SSLF groups is higher than that in the AH + SCP group (p = 0.047). There is no significant difference among three VH + SSLF, LH + SCP and AH + SCP groups regarding scores of POPDI-6, UDI-6, CRADI-8, and PISQ-12. CONCLUSION: In the present study, it was concluded that AH + SCP group had a higher quality of life than the VH + SSLF group did while the sexual function was not affected significantly by the vaginal or abdominal surgical procedures. The pelvic surgeon should skillfully choose different prolapse surgical techniques to tailor the surgical treatment to the patient's needs.

Virus Like Particles (VLP) as multivalent vaccine candidate against Chikungunya, Japanese Encephalitis, Yellow Fever and Zika Virus.

Mosquito borne viral diseases are an emerging threat as evident from the recent outbreak of Zika virus (ZIKV) as well as repeated outbreaks of Chikungunya (CHIKV), Yellow fever (YFV) and Japanese encephalitis (JEV) virus in different geographical regions. These four arboviruses are endemic in overlapping regions due to the co-prevalence of the transmitting mosquito vector species Aedes and Culex. Thus, a multivalent vaccine that targets all four viruses would be of benefit to regions of the world where these diseases are endemic. We developed a potential Virus Like Particle (VLP) based multivalent vaccine candidate to target these diseases by using stable cell lines that continuously secrete VLPs in the culture supernatants. Moreover, inclusion of Capsid in the VLPs provides an additional viral protein leading to an enhanced immune response as evident from our previous studies with ZIKV. Immunization of Balb/c mice with different combinations of Capsid protein containing VLPs either as monovalent, bivalent or tetravalent formulation resulted in generation of high levels of neutralizing antibodies. Interestingly, the potential tetravalent VLP vaccine candidate provided strong neutralizing antibody titers against all four viruses. The 293 T stable cell lines secreting VLPs were adapted to grow in suspension cultures to facilitate vaccine scale up. Our stable cell lines secreting individual VLPs provide a flexible yet scalable platform conveniently adaptable to different geographical regions as per the need. Further studies in appropriate animal models will be needed to define the efficacy of the multivalent vaccine candidate to protect against lethal virus challenge.

Capsid containing virus like particle vaccine against Zika virus made from a stable cell line.

Zika virus infection during pregnancy is associated with severe birth defects including microcephaly in the new born. The lack of specific treatment calls for the development of a safe and effective vaccine for use in pregnant women. We recently tested the efficacy of a Virus Like Particle (VLP) vaccine for Zika virus in mice and found that Capsid-preMembrane-Env (CprME) VLPs generated a better neutralizing antibody response than preMembrane-Env (prME) VLPs. The superiority of CprME VLPs suggested that inclusion of capsid in the vaccine may enhance the immune response. However, production of CprME VLPs requires co-expression of NS2B-3 protease, which creates a major hurdle for generation of stable cell lines. To overcome this limitation, we generated a bicistronic vector that expresses CprME and NS2B-3 using an IRES sequence. This bicistronic expression cassette, in a lentiviral vector, was used to create a stable cell line that constitutively secretes CprME VLPs. The expression of NS2B-3, presence of capsid in the secreted VLPs, efficiency of VLP release, and stability of the cell line was extensively tested. Antigen sparing studies in mice using prME and CprME VLPs, both derived from stable cell lines, confirmed the superiority of CprME VLPs in generation of neutralizing antibody response. Capsid specific antibodies were detected in CprME VLP immunized mice providing mechanistic insights into the superiority of these VLPs. Challenge of CprME VLP immunized mice with Zika PRVABC59 showed complete protection against day 3 viremia further validating the efficacy of the vaccine. Our study is the first to generate a stable cell line secreting Zika CprME VLPs via natural NS2B-3 cleavage, demonstrate incorporation of capsid in CprME VLPs and complete protection in challenge studies. This is a major advancement for the Zika vaccine platform that is safe for use in pregnant women and readily scalable for use in developing countries.

Differential Pathogenicity of SHIV KB9 and 89.6 Env Correlates with Bystander Apoptosis Induction in CD4+ T cells.

SHIV variants KB9 and 89.6 show differential pathogenesis in primate models with KB9 causing rapid CD4 decline while 89.6 failing to induce disease. We attempted to determine whether the differential pathogenicity of KB9 versus 89.6 was a result of differential bystander apoptosis inducing potential (AIP) of the Env glycoproteins from these viruses. We find that the KB9 Env was highly potent at inducing bystander apoptosis in CD4+ target cells compared to 89.6 Env. Cell death induction by KB9 showed classical signs of apoptosis including mitochondrial depolarization, caspase activation and PARP cleavage. Inhibiting Env mediated fusion by T20 peptide inhibited KB9 mediated bystander apoptosis. KB9 and 89.6 differed in terms of co-receptor usage with 89.6 preferring CXCR4 while KB9 using both CXCR4 and CCR5 with equal efficiency. Our study suggests that higher bystander AIP of KB9 Env compared to 89.6 may be the basis for the differential pathogenesis of these viruses.

Anti-Zika Activity of a Library of Synthetic Carbohydrate Receptors.

Zika virus (ZIKV), a mosquito-borne flavivirus, is a global health concern because of its association with severe neurological disorders. Currently, there are no antiviral therapies that have been specifically approved to treat ZIKV, and there is an urgent need to develop effective anti-ZIKV agents. Here, we report anti-ZIKV activity of 16 synthetic carbohydrate receptors (SCRs) that inhibit ZIKV infection in Vero and HeLa cells. Using a ZIKV reporter virus particle-based infection assay, our data demonstrates these SCRs are highly potent with IC50s as low as 0.16 µM and negligible toxicity at several-fold higher concentrations. Time-of-addition studies showed that these SCRs inhibit the early stages of the virus infection, which is consistent with the proposed mode of action, where the SCRs likely inhibit binding between the virus and cell-surface glycans, thereby preventing viral entry into the cells and, as such, this study demonstrates a potential new strategy against ZIKV.