Modulation of SLFN11 induces changes in DNA Damage response in breast cancer.

BACKGROUND: Lack of Schlafen family member 11 (SLFN11) expression has been recently identified as a dominant genomic determinant of response to DNA damaging agents in numerous cancer types. Thus, several strategies aimed at increasing SLFN11 are explored to restore chemosensitivity of refractory cancers. In this study, we examined various approaches to elevate SLFN11 expression in breast cancer cellular models and confirmed a corresponding increase in chemosensitivity with using the most successful efficient one. As oncogenic transcriptomic downregulation is often driven by methylation of the promotor region, we explore the demethylation effect of 5-aza-2'-deoxycytidine (decitabine), on the SLFN11 gene. Since SLFN11 has been reported as an interferon inducible gene, and interferon is secreted during an active anti-tumor immune response, we investigated the in vitro effect of IFN-γ on SLFN11 expression in breast cancer cell lines. As a secondary approach to pick up cross talk between immune cells and SLFN11 expression we used indirect co-culture of breast cancer cells with activated PBMCs and evaluated if this can drive SLFN11 upregulation. Finally, as a definitive and specific way to modulate SLFN11 expression we implemented SLFN11 dCas9 (dead CRISPR associated protein 9) systems to specifically increase or decrease SLFN11 expression. RESULTS: After confirming the previously reported correlation between methylation of SLFN11 promoter and its expression across multiple cell lines, we showed in-vitro that decitabine and IFN-γ could increase moderately the expression of SLFN11 in both BT-549 and T47D cell lines. The use of a CRISPR-dCas9 UNISAM and KRAB system could increase or decrease SLFN11 expression significantly (up to fivefold), stably and specifically in BT-549 and T47D cancer cell lines. We then used the modified cell lines to quantify the alteration in chemo sensitivity of those cells to treatment with DNA Damaging Agents (DDAs) such as Cisplatin and Epirubicin or DNA Damage Response (DDRs) drugs like Olaparib. RNAseq was used to elucidate the mechanisms of action affected by the alteration in SLFN11 expression. In cell lines with robust SLFN11 promoter methylation such as MDA-MB-231, no SLFN11 expression could be induced by any approach. CONCLUSION: To our knowledge this is the first report of the stable non-lethal increase of SLFN11 expression in a cancer cell line. Our results show that induction of SLFN11 expression can enhance DDA and DDR sensitivity in breast cancer cells and dCas9 systems may represent a novel approach to increase SLFN11 and achieve higher sensitivity to chemotherapeutic agents, improving outcome or decreasing required drug concentrations. SLFN11-targeting therapies might be explored pre-clinically to develop personalized approaches.

Knowledge, Awareness, and Perception of Common Eye Diseases and Eye Donation Among People Seeking Healthcare in a Tertiary Hospital in Telangana, South India.

Background Globally billions of people have vision impairment (VI) or blindness, and at least half of the VI could have been prevented or has yet to be addressed. With the policies focused exclusively on treating ailments, we need to recognize the need to educate the country's population regarding diseases and their outcomes. This is evident in the poor eye donation rates, as documented by the Eye Bank Association of India (EBAI). The National Programme for Prevention of Blindness (NPCB) also advocates the need for active campaigning to promote eye donations and improve the corneal procurement rate by increasing health awareness. This study aimed to assess the knowledge, awareness, and perception regarding eye diseases and eye donation among the rural population of Telangana, South India. Methods A cross-sectional study involving 150 participants who were randomly selected from non-triaged attendants in the outpatient queue at Mahavir institute of medical sciences (MIMS) was included. Trained enumerators used verbally administered, semi-structured questionnaires on their awareness and knowledge of cataracts, glaucoma, diabetic retinopathy, and night blindness. The Chi-square test was applied to determine the statistical significance of the results obtained from the pre-test and post-test. The significance threshold of the p-value was set at <0.05. Results The population studied belonged to a mean age of 34.98 years. The study included 72 (48%) male and 78 (52%) female subjects, and the majority (48.6%) of them belonged to the age range of 21-30 years. More than 85% of subjects belonged to the middle and lower middle class, and the majority (74.7%) were Hindus by religion. Regarding cataracts, 64 (42.7%) did not know, and 86 (57.3%) had varied perceptions. The awareness regarding glaucoma was the least (88.7%) of all common ocular diseases. The study showed a significant association between knowledge of eye diseases and literacy status plus the population's socioeconomic status (p<0.05 ). A significant association was found to exist between willingness to eye donation and the religion of the study population (p<0.05). Conclusion This study identifies that the awareness regarding various ocular diseases was poor. Moreover, the participants had an alarmingly high misconception regarding different aspects of eye donation. Increasing public awareness is essential to minimize eye diseases, improve eye care, and encourage eye donations.

World Health Organization Versus Diabetes in Pregnancy Study Group India Approaches in the Screening of Gestational Diabetes Among Pregnant Women With Risk Factors: A Study Among Rural Population of Telangana, South India.

Background Gestational diabetes mellitus (GDM) represents a pathological condition wherein pregnant women (PW) suffer from glycemic dysregulation, which predisposes them to an increased risk of developing complications related to pregnancy and childbirth. The most commonly used guidelines to screen for GDM include those provided by the World Health Organization (WHO), the American Congress of Obstetricians and Gynecologists, the Canadian Diabetes Association, and the International Association of Diabetes and Pregnancy Study Group. The Diabetes in Pregnancy Study Group India (DIPSI) guidelines are national-level recommendations to screen for GDM in India. This study aimed to compare the efficacy of DIPSI criteria versus the WHO guidelines in screening for GDM among the rural population of Telangana, South India Methods A total of 300 PW aged 19-35 years with a gestational age of 24-28 weeks attending the antenatal clinic attached to Mahavir Institute of Medical Sciences (MIMS), Vikarabad, Telangana, India were included in the study. The study was approved by the Institutional Ethics Committee of MIMS, and informed consent was obtained from all the participants. Of the 300 subjects included, 75 PW were categorized as at-risk for GDM based on risk factors and were included for further analysis. The data relating to body mass index (BMI), oral glucose tolerance test, and the diagnosis of GDM based on DIPSI and the WHO criteria were collected. Results Out of the 75 PW included in the study, an overall GDM prevalence of 32% was noted among which 20 (26.7%) were diagnosed using the WHO criteria, 12 (16%) by DIPSI criteria, and the remaining 73.3% were non-GDM women. The mean gestational age and BMI among non-GDM and GDM patients were 24.74±4.15 weeks, 22.24±3.60 kg/m2, and 25.70±4.40, 24.48±3.37 kg/m2 (p<0.01), respectively. The activities of glucose at the second hour after a GTT among non-GDM and GDM cases were 113.70±20.4 mg/dL and 128.04±18.6 mg/dL (p=0.004), respectively.  Conclusion DIPSI criteria could identify fewer numbers of GDM women as compared to the WHO criteria. Although the DIPSI criteria are convenient and prescribe less number of interventions, they could possibly miss many cases of GDM. Moreover, PW who remain undiagnosed could, in the future, be at risk of developing diabetes. Based on the study results and because risks should outweigh the benefits, we propose that DIPSI cannot be implemented as a single criterion to screen for GDM among PW in Indian settings.

Fiber-enriched botanicals: A therapeutic tool against certain metabolic ailments.

Plant-based foods are natural sources including vegetables, fruits, cereals and legumes. These foods consist of various types of nutrients in which carbohydrate is the basic component. However, some plant-based diets contain carbohydrates in the form of fiber. The fiber is usually a nondigestible polysaccharide that is not digested in the human body. It is present in the form of soluble or insoluble in different part of foods like peel, bran, pulp and grain. Pectin, beta-glucan, mucilage, psyllium, resistant starch and inulin are soluble fiber, and cellulose, hemicellulose and lignin are insoluble fiber attained from plant foods. The major function enhances immunity by creating gastrointestinal barrier, mucus production, immune cell activity and IgA level. Previous evidences showed that peoples with strong immunity have fewer chances of viral disease. A recent viral disease named COVID-19 spread in the world and millions of peoples died due to this viral disease. Coronavirus mostly attacks humans that suffer with weak immune system. It is due chronic diseases like diabetes and CVD (cardiovascular disease). The current review shows that fiber-containing plant-based foods boost immunity and aid human against COVID-19. The therapeutic role of fiber in the human body is to control the risk of hypertension and diabetes because a high-fiber diet has the ability to lower cholesterol, blood pressure and blood sugar. Fibers aid in GIT (gastrointestinal tract) and prevent constipation because it absorbs water and adds bulk to stool.

Assessment of Breastfeeding, Weaning, and Complementary Feeding Practices Among Women Attending a Tertiary Care Teaching Hospital in South India.

Introduction Infants depend exclusively on mothers' milk for nutrition in their early months of life. However, some infants are given cow/buffalo milk during insufficiency. After six months, the children are supplemented with complementary food, including solid food, to meet the nutritional requirements of a growing infant, a process known as weaning. Therefore, it is important for mothers to have a clear understanding of the nature of food, and the nutritional requirements of the children. This study aimed to understand the knowledge, awareness, and practice of breastfeeding, weaning, and complementary feeding among women. Methods A cross-sectional questionnaire-based observational study was carried out between May and June 2021. The study included 150 women attending the outpatient department of pediatrics attached to Mahavir Institute of Medical Sciences (MIMS), Vikarabad, Telangana, India. After taking the informed consent, the researcher filled out a pre-validated questionnaire based on the subjects' responses. The data regarding socio-demographic details and information regarding knowledge, timing, practices of weaning, knowledge of nutritional requirements, and child feeding practices were collected as a pre-test. An educational briefing of the essentials of child feeding practices, weaning, and dietary requirements was undertaken using chalk and board and audio-visual aids, among others. In the post-test, the knowledge of mothers regarding weaning practices, the importance of weaning, and understanding of the nutritional requirements and their inclusion in a child's diet was assessed. The quantitative data were represented as percentages. The Chi-square test was applied to find out the statistical significance of the results obtained from the study. Results Of the 150 women who participated in the study, the majority belonged to the age group of 18-23 years (66.7%). Most women were illiterate (34%) and only 12% were graduates. More than 70% of the participants belonged to the middle and lower middle class. The majority of participants (96.7%) had carried out exclusive breastfeeding for the first six months, and 63.35% of mothers had initiated weaning their child between the age of 6-12 months. Only 37.4% of mothers started breastfeeding within hours after the delivery. A combination of all foods (36%), rice/wheat (29%), processed food (18.8%), fruits and vegetables (9.2%), and pulses (6.7%) were chosen for weaning. Many felt that eggs and meat supplementation can be done between 12-18 months. Conclusion The knowledge of breastfeeding and best practices of weaning and complementary feeding habits significantly affects the child's growth and overall health. The women in this study had a moderate level of understanding regarding the aspects of breastfeeding, weaning practices, and complementary dietary requirements during weaning.

Polycystic Ovarian Syndrome: Prevalence, Predisposing Factors, and Awareness Among Adolescent and Young Girls of South India.

Introduction Polycystic ovarian syndrome (PCOS) is a leading cause of infertility among women throughout the world. PCOS is an endocrine abnormality that presents as hyperandrogenemia, anovulation, and/or polycystic ovaries. The exact causes of PCOS are not entirely understood. However, PCOS may be hereditary and associated with abnormalities such as high body mass index (BMI) and obesity, among others. This study is carried out to assess the prevalence and awareness among adolescents and young girls belonging to South India. Methods This was a prospective study that included 250 adolescent and young girls aged between 13 and 25 years. Demographic details including age, educational status, and family income were collected from all the participants after obtaining informed consent. BMI was measured in all the participants. A pre-test was conducted to assess the level of awareness followed by a 30-minute informative briefing on PCOS. All participants were asked to fill up a post-briefing questionnaire. The data collected were processed using statistical software SPSS 11.0, and the chi-square test was applied to find out the significance of the results. Results Of the total 250 participants included, the mean age was 16.96 years and most participants (78%) belonged to the age group of 13 to 19 years. Most (78%) of the study participants had normal BMI (18-24.9 kg/m2), 17.6% were underweight (BMI < 18), and 4.4% were overweight (BMI > 25). A PCOS prevalence rate of 6.8% was noted among the study participants. A majority (78.4%) of the study participants were unaware of PCOS, and 6.8% were being treated for PCOS. The source of knowledge of PCOS was majorly teachers (37%), followed by doctors (31.5%), the internet (11%), and friends (7.5%). Lack of information and publicity (63%) were found to be the most significant reason for low levels of awareness. Conclusion PCOS is a common health problem among adolescents and young girls. Most study participants were unaware of the symptoms and management of the condition. Therefore, the disease burden noted in this study does not necessarily depict the real prevalence. Increasing awareness programs will facilitate improved understanding, increased diagnoses, and effective management of PCOS.

Child with intellectual disability misdiagnosed as autism: A case study.

Misdiagnosis has become a public health crisis and it is unavoidable in all health care systems. This case study is about a nine year old boy, referred by the special school teacher presenting with the complaints of Autism and slow learning. Contrary to his complaints, the developmental assessment showed that the child had no symptoms of Autism, whereas moderate symptoms of Intellectual Disability were made known. The brief therapeutic plan focused on enhancing his functional level of adaptive behaviour. This case report holds its implication to drift attention of stakeholders working in different health care systems towards the issue of misdiagnosis.

Toxicity of Five Plant Volatiles to Adult and Egg Stages of Drosophila suzukii Matsumura (Diptera: Drosophilidae), the Spotted-Wing Drosophila.

The environmental impact of methyl bromide (MB) has resulted in its phase out as an insecticidal fumigant except for critical use exempted categories. Consequently, there is an urgent need to develop an environmentally sustainable MB alternative fumigant. trans-Cinnamaldehyde (TC), benzaldehyde, allyl isothiocyanate (AITC), hexanal, and ethyl formate (EF) are naturally occurring plant volatiles with insecticidal properties. This study assessed the toxicity of these plant volatiles to adult and egg stages of the spotted-wing drosophila (SWD) (Drosophila suzukii Matsumura). The plant volatile treatments had a significant effect on adult SWD mortality. The descending order of toxicity to adult SWD was benzaldehyde > AITC > TC > hexanal > EF at a headspace concentration of 0.50 µL/L air for 24 h. All the volatiles, at a concentration of 4.00 µL/L air, significantly inhibited larval emergence from SWD eggs in artificial diet compared to the control. At a 0.50 µL/L air level, among the volatiles tested, only AITC exhibited 100% inhibition against larval emergence from SWD eggs in blueberry fruits after 24 h exposure. In summary, this study shows that all volatiles tested elicited varying degrees of toxicity toward SWD adults and eggs. However, AITC was the most efficacious volatile and the one with the greatest promise as a post-harvest fumigant for both adult and egg stages of SWD.

High-risk human papillomaviruses and Epstein-Barr virus in breast cancer in Lebanese women and their association with tumor grade: a molecular and tissue microarray study.

BACKGROUND: High-risk human papillomaviruses (HPVs) are present and can cooperate with Epstein-Barr virus (EBV) to initiate and/or enhance the progression of several types of human carcinomas including cervical as well as head and neck; in parallel, it has been recently pointed out that these oncoviruses can be detected in human breast cancers. Thus, we herein explored the presence/co-presence of high-risk HPVs and EBV in breast cancer in Lebanese women. METHODS: A cohort of 102 breast cancer samples and 14 normal breast tissues were assessed for the presence of HPVs and EBV. Polymerase chain reaction (PCR) and immunohistochemistry (IHC) analysis in addition to tissue microarray (TMA) platform were used in this study. RESULTS: We found the presence of HPV in 66/102 (65%) of our samples, while EBV is present in 41/102 (40%) of the cohort. Additionally, our data showed that high-risk HPV types (52, 35, 58, 45, 16 and 51) are the most frequent in breast cancer in Lebanese women. Meanwhile, we report that high-risk HPVs and EBV are co-present in 30/102 (29%) of the samples; more significantly, our results indicate that their co-presence is associated with tumor grade (p = 0.03). CONCLUSION: Our data revealed that HPVs and EBV are present/co-present in human breast cancer where they may play an important role in its development and/or progression; thus, we believe that further investigations are essential to confirm and elucidate the presence/co-presence of these oncoviruses and the underlying mechanisms of their interaction in breast carcinogenesis.

Oncoproteins of High-Risk HPV and EBV Cooperate to Enhance Cell Motility and Invasion of Human Breast Cancer Cells via Erk1/Erk2 and ß-Catenin Signaling Pathways.

Breast cancer is a leading cause of death in women around the world. Most breast cancer-related deaths are a result of complications from the metastatic spread. Several recent studies reported that high-risk human papillomaviruses (HPVs) and Epstein-Barr virus (EBV) are co-presented in different types of human carcinomas including breast; however, the cooperative effects between high-risk HPVs and EBV oncoproteins in human breast cancer have not been investigated yet. Thus, we herein explored the cooperation outcome between E6/E7 and latent membrane protein 1 (LMP1) oncoproteins of high-risk HPV type 16 and EBV, respectively, in two human breast cancer cell lines, MCF7 and MDA-MB-231. Our data revealed that the cooperation of E6/E7 and LMP1 oncoproteins stimulates cell proliferation and deregulates cell cycle progression of human breast cancer and normal mammary cells; in parallel, we noted that E6/E7/LMP1 incite colony formation of both breast cancer cell lines but not normal cells. More significantly, our results point out that the co-expression of E6/E7 and LMP1 oncoproteins enhances cell motility and invasion of MCF7 and MDA-MB-231 cell lines; this is accompanied by deregulation of epithelial-mesenchymal transition biomarkers including E-cadherin, ß-catenin, fascin, and vimentin. The molecular pathway analysis of HPV and EBV oncoproteins cooperation shows that it can enhance the phosphorylation of extracellular signal-regulated kinases (Erk1/Erk2) in addition to ß-catenin, which could be behind the effect of this cooperation in our cell models. The study clearly suggests that high-risk HPV and EBV coinfection can play an important role in breast cancer progression via Erk1/Erk2 and ß-catenin signaling pathways.

Psychosocial Reactions of Pakistani Students Towards COVID-19: A Prevalence Study.

BACKGROUND: The current study aimed at investigating the manifestation and prevalence of the commonly reported psychosocial reactions in the university students following the onset of COVID-19 pandemic. METHODS: This cross-sectional survey was carried out during April to May, 2020 in the city of Lahore. Based on the interviews and presenting problems of thirty-nine self-referred students to student counselling service center, a list of twenty-seven self-report measure was presented through an online cross-sectional survey of 510 students enrolled in a private institute. The age range of the participants was between 17-26 years (M, 21.86; SD, 2.94). RESULTS: The results showed that the most frequently reported reactions by university students during COVID-19 is restricted daily routine (92%), preoccupation with cleanliness (86%), feeling uncertain about future (85%), feeling bored (84%) and low mood (84%). The least frequently reported reaction was financial crisis (48%). The findings further showed that 18% of the participants had mild, 34% had moderate, 29% severe and 19% very severe level of problems. CONCLUSIONS: The finding revealed that university students have significantly affected by this pandemic which need attention from mental health professionals.

Dasatinib and PD-L1 inhibitors provoke toxicity and inhibit angiogenesis in the embryo.

Dasatinib is a targeted cancer therapy, while programmed death ligand 1 (PD-L1) inhibitors are a form of immune checkpoint therapy used to treat various types of cancers. Several studies showed the potential efficacy of these drugs in the management of triple-negative breast cancer- an aggressive subtype of breast cancer, which can develop during pregnancy. Nevertheless, side effects of Dasatinib (DA) and PD-L1 drugs during pregnancy, especially in the early stages of embryogenesis are not explored yet. The aim of this study is to assess the individual and combined toxicity of DA and PD-L1 inhibitors during the early stages of embryogenesis and to evaluate their effect(s) on angiogenesis using the chorioallantoic membrane (CAM) model of the embryo. Our results show that embryos die at greater rates after exposure to DA and PD-L1 inhibitors as compared to their matched controls. Moreover, treatment with these drugs significantly inhibits angiogenesis of the CAM. To further elucidate key regulator genes of embryotoxicity induced by the actions of PD-L1 and DA, an RT-PCR analysis was performed for seven target genes that regulate cell proliferation, angiogenesis, and survival (ATF3, FOXA2, MAPRE2, RIPK1, INHBA, SERPINA4, and VEGFC). Our data revealed that these genes are significantly deregulated in the brain, heart, and liver tissues of exposed embryos, compared to matched control tissues. Nevertheless, further studies are necessary to evaluate the effects of these anti breast cancer drugs and elucidate their role during pregnancy.

The co-presence of high-risk human papillomaviruses and Epstein-Barr virus is linked with tumor grade and stage in Qatari women with breast cancer.

High-risk human papillomaviruses (HPV) can be present and cooperate with Epstein-Barr virus (EBV) to promote the onset and/or progression of various cancers including cervical, breast, head and neck as well as colorectal. In this investigation, we explored the co-prevalence of high-risk HPV and EBV in 74 breast cancer tissues from Qatari women using polymerase chain reaction. We found that high-risk HPV and EBV are present in 48/74 (65%) and 36/74 (49%) of the cases, respectively. While we noted that the presence of HPV presence is associated with triple-negative breast cancer (TNBC) (p = .008), however, the presence of EBV did not correlate with any breast cancer subgroup. Moreover, our data revealed that high-risk HPV and EBV are co-present in 35/74 (47%) of the samples and their co-presence is significantly associated with tumor grade (p = .04) and tumor stage (p = .04). These data indicate that HPV and EBV are commonly co-present in breast cancer and their association could be linked with a more aggressive tumor phenotype. Thus, further investigations are essential to understand the underlying mechanisms of HPV and EBV cooperation in breast carcinogenesis.

Role of Purine-Rich Regions in Mason-Pfizer Monkey Virus (MPMV) Genomic RNA Packaging and Propagation.

A distinguishing feature of the Mason-Pfizer monkey virus (MPMV) packaging signal RNA secondary structure is a single-stranded purine-rich sequence (ssPurines) in close vicinity to a palindromic stem loop (Pal SL) that functions as MPMV dimerization initiation site (DIS). However, unlike other retroviruses, MPMV contains a partially base-paired repeat sequence of ssPurines (bpPurines) in the adjacent region. Both purine-rich sequences have earlier been proposed to act as potentially redundant Gag binding sites to initiate the process of MPMV genomic RNA (gRNA) packaging. The objective of this study was to investigate the biological significance of ssPurines and bpPurines in MPMV gRNA packaging by systematic mutational and biochemical probing analyses. Deletion of either ssPurines or bpPurines individually had no significant effect on MPMV gRNA packaging, but it was severely compromised when both sequences were deleted simultaneously. Selective 2' hydroxyl acylation analyzed by primer extension (SHAPE) analysis of the mutant RNAs revealed only mild effects on structure by deletion of either ssPurines or bpPurines, while the structure was dramatically affected by the two simultaneous deletions. This suggests that ssPurines and bpPurines play a redundant role in MPMV gRNA packaging, probably as Gag binding sites to facilitate gRNA capture and encapsidation. Interestingly, the deletion of bpPurines revealed an additional severe defect on RNA propagation that was independent of the presence or absence of ssPurines or the gRNA structure of the region. These findings further suggest that the bpPurines play an additional role in the early steps of MPMV replication cycle that is yet to be identified.

Elaeagnus angustifolia Plant Extract Inhibits Epithelial-Mesenchymal Transition and Induces Apoptosis via HER2 Inactivation and JNK Pathway in HER2-Positive Breast Cancer Cells.

Elaeagnus angustifolia (EA) is a medicinal plant used for treating several human diseases in the Middle East. Meanwhile, the outcome of EA extract on HER2-positive breast cancer remains nascent. Thus, we herein investigated the effects of the aqueous EA extract obtained from the flowers of EA on two HER2-positive breast cancer cell lines, SKBR3 and ZR75-1. Our data revealed that EA extract inhibits cell proliferation and deregulates cell-cycle progression of these two cancer cell lines. EA extract also prevents the progression of epithelial-mesenchymal transition (EMT), an important event for cancer invasion and metastasis; this is accompanied by upregulations of E-cadherin and ß-catenin, in addition to downregulations of vimentin and fascin, which are major markers of EMT. Thus, EA extract causes a drastic decrease in cell invasion ability of SKBR3 and ZR75-1 cancer cells. Additionally, we found that EA extract inhibits colony formation of both cell lines in comparison with their matched control. The molecular pathway analysis of HER2 and JNK1/2/3 of EA extract exposed cells revealed that it can block HER2 and JNK1/2/3 activities, which could be the major molecular pathway behind these events. Our findings implicate that EA extract may possess chemo-preventive effects against HER2-positive breast cancer via HER2 inactivation and specifically JNK1/2/3 signaling pathways.

Co-presence of human papillomaviruses and Epstein-Barr virus is linked with advanced tumor stage: a tissue microarray study in head and neck cancer patients.

BACKGROUND: Human papillomaviruses (HPVs) and Epstein-Barr virus (EBV), known oncoviruses, can be co-present and cooperate in the initiation and/or progression of human carcinomas, including head and neck. Based on this fact, we recently reported the prevalence of both HPVs and EBV in cervical and breast cancers. METHODS: We herein explore for the first time the co-prevalence of high-risk HPVs and EBV in 98 head and neck (HN) squamous cell carcinoma (SCC) tissues from Bosnian patients using polymerase chain reaction (PCR) and immunohistochemistry (IHC) analysis, as well as tissue microarray methodology. RESULTS: The majority of these cancer tissue cases were from the oral cavity (68%). We found that high-risk HPVs and EBV are co-present in 34.7% of the SCC samples; with a significant correlation between the various HPV types and EBV co-incidence (p = 0.03). Our data showed that 30.8% of oral SCCs are positive for E6 oncoprotein of high-risk HPVs and 44.6% are positive for LMP1 of EBV. The most commonly expressed HPVs in our HNSCC samples include HPV types 16, 18, 45 and 58. Additionally, 37.5% of oral SCCs are positive for both HPVs and EBV, with statistically significant association between high-risk HPV types and EBV (p < 0.05). More importantly, our data revealed that the co-presence of HPV and EBV is strongly correlated with advanced tumor stage (p = 0.035). CONCLUSION: In this study we show that HPV and EBV oncoviruses are co-present in HNSCC, particularly in oral cancer, where they can cooperate in the initiation and/or progression of this cancer. Thus, further studies are necessary to elucidate the mechanism of this cooperation.

Anti-hypertrophic effect of Na+/H+ exchanger-1 inhibition is mediated by reduced cathepsin B.

Previous studies have established the role of Na+/H+ exchanger isoform-1 (NHE1) and cathepsin B (Cat B) in the development of cardiomyocyte hypertrophy (CH). Both NHE1 and Cat B are activated under acidic conditions suggesting that their activities might be interrelated. The inhibition of NHE1 has been demonstrated to reduce cardiac hypertrophy but the mechanism that contributes to the anti-hypertrophic effect of NHE1 inhibition still remains unclear. H9c2 cardiomyoblasts were stimulated with Angiotensin (Ang) II in the presence and absence of N-[2-methyl-4,5-bis(methylsulphonyl)-benzoyl]-guanidine, hydrochloride (EMD, EMD 87580), an NHE1 inhibitor or CA-074Me, a Cat B inhibitor, and various cardiac hypertrophic parameters, namely cell surface area, protein content and atrial natriuretic peptide (ANP) mRNA were analyzed. EMD significantly suppressed markers of cardiomyocyte hypertrophy and inhibited Ang II stimulated Cat B protein and gene expression. Cat B is located within the acidic environment of lysosomes. Cat B proteases are released into the cytoplasm upon disintegration of the lysosomes. EMD or CA-074Me prevented the dispersal of the lysosomes induced by Ang II and reduced the ratio of LC3-II to LC3-I, a marker of autophagy. Moreover, Cat B protein expression and MMP-9 activity in the extracellular space were significantly attenuated in the presence of EMD or CA-074Me. Our study demonstrates a novel mechanism for attenuation of the hypertrophic phenotype by NHE1 inhibition that is mediated by a regression in Cat B. The inhibition of Cat B via EMD or CA-074Me attenuates the autosomal-lysosomal pathway and MMP-9 activation.

High-Risk Human Papillomaviruses and Epstein-Barr Virus in Colorectal Cancer and Their Association with Clinicopathological Status.

Colorectal cancer (CRC) is a common malignancy with a high mortality rate worldwide. It is a complex, multifactorial disease that is strongly impacted by both hereditary and environmental factors. The role of microbes (e.g., viruses) in the pathogenesis of CRC is poorly understood. In the current study, we explored the status of high-risk human papillomaviruses (HPV) and Epstein-Barr virus (EBV) in a well-defined CRC cohort using immunohistochemistry and polymerase chain reaction assays. Our data showed that high-risk HPVs were common (~80%) and EBV had a low presence (14-25%) in the CRC samples. The most common high-risk HPVs are HPV16, 31, 18, 51, 52 and 45 genotypes. The co-presence of high-risk HPV and EBV was observed in ~16% of the sample population without any significant association with the clinicopathological variables. We conclude that high-risk HPVs are very prevalent in CRC samples while EBV positivity is relatively low. The co-expression of the two viruses was observed in a minority of cases and without any correlation with the studied parameters. Further studies are necessary to confirm the clinical relevance and potential therapeutic (preventive) effects of the observations reported herein.

Co-prevalence of human Papillomaviruses (HPV) and Epstein-Barr virus (EBV) in healthy blood donors from diverse nationalities in Qatar.

BACKGROUND: Infections by both human oncoviruses, human Papillomaviruses (HPV) and Epstein-Barr virus (EBV) are very common in the adult human population and are associated with various malignancies. While HPV is generally transmitted sexually or via skin-to-skin contact, EBV is frequently transmitted by oral secretions, blood transfusions and organ transplants. This study aims to determine the prevalence and circulating genotypes of HPV and EBV in healthy blood donors in Qatar. METHODS: We explored the co-prevalence of high-risk HPVs and EBV in 378 males and only 7 females blood donors of different nationalities (mainly from Qatar, Egypt, Syria, Jordan, Pakistan, and India) residing in Qatar, using polymerase chain reaction (PCR). DNA was extracted from the buffy coat and genotyping was performed using PCR and nested-PCR targeting E6 and E7 as well as LMP-1 of HPV and EBV, respectively. RESULTS: We found that from the total number of 385 cases of healthy blood donors studied, 54.8% and 61% of the samples are HPVs and EBV positive, respectively. Additionally, our data revealed that the co-presence of both high-risk HPVs and EBV is 40.4% of the total samples. More significantly, this study pointed out for the first time that the most frequent high-risk HPV types in Qatar are 59 (54.8%), 31 (53.7%), 52 (49.1%), 51 (48.6%), 58 (47%) and 35 (45.5%), while the most commonly expressed low-risk HPV types are 53 (50.6%), 11 (45.5), 73 (41.7%) and 6 (41.3%), with all the cases showing multiple HPVs infection. CONCLUSION: In this study, we demonstrated for the first time that HPV and EBV are commonly co-present in healthy blood donors in Qatar. On the other hand, it is important to highlight that these oncoviruses can also be co-present in several types of human cancers where they can cooperate in the initiation and/or progression of these cancers. Therefore, more studies regarding the co-presence of these oncoviruses and their interaction are necessary to understand their cooperative role in human diseases.