Breast cancer is characterized by considerable metabolic diversity. A relatively high percentage of patients diagnosed with breast carcinoma do not respond to standard-of-care treatment, and alteration in metabolic pathways nowadays is considered one of the major mechanisms responsible for therapeutic resistance. Consequently, there is an emerging need to understand how metabolism shapes therapy response, therapy resistance and not ultimately to analyze the metabolic changes occurring after different treatment regimens. The most commonly applied neoadjuvant chemotherapy regimens in breast cancer contain an anthracycline (doxorubicin or epirubicin) in combination or sequentially administered with taxanes (paclitaxel or docetaxel). Despite several efforts, drug resistance is still frequent in many types of breast cancer, decreasing patients’ survival. Understanding how tumor cells rapidly rewire their signaling pathways to persist after neoadjuvant cancer treatment have to be analyzed in detail and in a more complex system to enable scientists to design novel treatment strategies that target different aspects of tumor cells and tumor resistance. Tumor heterogeneity, the rapidly changing environmental context, differences in nutrient use among different cell types, the cooperative or competitive relationships between cells pose additional challenges in profound analyzes of metabolic changes in different breast carcinoma subtypes and treatment protocols. Delineating the contribution of metabolic pathways to tumor differentiation, progression, and resistance to different drugs is also the focus of research. The present review discusses the changes in glucose and fatty acid pathways associated with the most frequently applied chemotherapeutic drugs in breast cancer, as well the underlying molecular mechanisms and corresponding novel therapeutic strategies. Whenoutcomes , 2.Since the first clinical trial  from 1997 that compared the prognostic value of neoadjuvant and adjuvant use of the same chemotherapy regimen, results of several other clinical trials have been published reporting no significant differences in the long-term outcomes for early-stage and locally advanced breast cancer with either approach , 4. MeanPersonalized therapeutical approaches in both the neoadjuvant and adjuvant setting play more and more important role in breast cancer management. Adding immunotherapies [anti-PD-L1  or anti-PD-1 (pembrolizumab)] or PARP-inhibitor  to standard chemotherapy may improve the rate of pCR in TNBC and women with germline BRCA mutations , 12.The classification of most frequently used chemotherapeutic drugs in neoadjuvant and adjuvant settings - mostly DNA alkylating, antimicrotubule, immunologic and hormonal agents, and antimetabolites - are presented in Considering the breast carcinoma subtypes in the UK the most commonly prescribed neoadjuvant chemotherapy regimes in breast cancers are: AC-containing combinations of fluorouracil, epirubicin and cyclophosphamide (FEC) with docetaxel/trastuzumab/pertuzumab for HER2-positive disease and FEC-docetaxel for HER2-negative disease  and epirubicin  and toxicity  have to be considered in each patient before prescription , 17.Several trials were and are designed to compare the effectiveness between ACs and/or taxane as neoadjuvant chemotherapy , 18 and The Collaborative Trials in Neoadjuvant Breast Cancer group analyzing the pCR association with long-term outcome have found that the highest pCR was achieved in HER2-positive/HR-negative breast cancer (50.3%) after administration of trastuzumab followed by TNBC (33.6%) and by grade 3 HR-positive/HER2-negative breast cancer with a pCR rate of 16.2%)  and docetaxel (DTX)) suppress microtubules involved in a variety of cellular processes like cell division, signaling, migration by binding to the β-subunit of the tubulin heterodimer  as a novel taxane drugwas developed and approved by the FDA in 2010 and used as a second-line treatment for metastatic castration-resistant prostate cancer patients .Though the initial response to taxanes is impressive, the resistance to these drugs is frequently observed and is supposed to be coordinated by: 1. alterations in tubulin, 2. proteins that interact with microtubules, 3. efflux pumps, 4. apoptotic proteins, 5. signal transduction pathways, 6. tumor metabolism. Accordingly, strategies to overcome resistance to PTX and DTX include inhibition of the efflux pumps, the use of novel taxane not interacting with drug efflux pumps, regulation of apoptotic and signal transduction pathways, and inhibition of metabolic pathways involved in drug resistance , 29. TubHow the sequence order of ACs and taxanes used in neoadjuvant settings influence the pCR rate is under considerable debate. A recent study showed that sequence order did not influence the primary endpoint of pCR rate (19% for AC-Taxane vs. 21% for Taxane-AC) and pCR rate was higher in patients with TNBC (32% vs. 13% in hormone-positive cancers) .Our knowledge about the impact of different AC-derived therapeutic drugs on the metabolic pathways in different cancers is mostly unknown. Achkar IW et al. in a very impressive study have analyzed the metabolic pathways affected by DOX treatment in chicken embryo tumor models (in ovo). By using two mass-spectrometry based platforms  they finally have found that after DOX treatment statistically significant alteration in 127 metabolites  can be documented on HD4 platform. Whereas on lipid platform they observed statistically significant decrease in 96 lipid molecules. Their results clearly indicate that DOX treatment generates metabolic rewiring .Related to paclitaxel a study performed on human breast carcinoma cell lines demonstrated significant differences in 31 metabolites such as fructose-6-phosphate, citric acid, glycerophosphoinositol and glycerol 3-phosphate when compared metabolic alterations in paclitaxel treated MCF-7 cell lines to untreated ones .Dysregulated cancer metabolism has been increasingly associated with acquired resistance to chemotherapeutic treatment, but no clear data are presented about how metabolic pathways are activated following oncological treatment. In addition to glycolysis that remains favored in resistant cancers undergoing AC treatment, experimental evidences also involve pentose phosphate pathway, fatty acid metabolism, nucleotide synthesis etc. However, there is an urgent need to analyze the complex system of tumor metabolism as the most pieces of evidence are often based on a single pathway without alternative explanation , 36.Beside dysregulated metabolism observed in several cancer types, metabolic reprogramming and metabolic heterogeneity play critical role in carcinogenesis and in acquired resistance to different chemotherapeutic agents. As cancer cells with different metabolic profiles may not respond similarly to anticancer treatment current studies suggest that metabolic heterogeneity needs to be integrated in routine practice to precisely predict breast cancer response to different treatments . It is aOf the relatively few studies demonstrating that chemotherapy agents used in the current treatment regimens in breast carcinomas cause metabolic reprogramming in cancer cells Desbats MA et al. presented that ACs induce increased glycolysis, GLUT1 and glutaminase level whereas taxanes induce elevation in glycolysis, in lactate dehydrogenase A (LDHA), pyruvate kinase M (PKM2), pyruvate dehydrogenase kinase 2 (PDK2) (inhibitor of pyruvate dehydrogenase complex), glutamine uptake, fatty acid synthase (FASN), mitophagy, mitochondrial mass and oxidative phosphorylation (OXPHOS) .Glycolysis is the major pathway of glucose metabolism occurring in the cytosol of the cells by which glucose is metabolized to pyruvate. Considering the aerobic and anaerobic conditions occurring in tumors and different organisms pyruvate transportation may follow two different pathways   and generates NADH and FADHvalents) . These r+ (the primary oxidizing agent of glycolysis) from NADH  converts pyruvate to lactate. This reaction also allows for the regeneration of NADrom NADH .Of the other pathways of carbohydrate metabolism  the PPP pathway was described as highly affected in different tumor types .Glycolysis is controlled by the following ten enzymes and enzymatic reactions   are present in the small intestine and the kidney, while passive transport occurs via ubiquitous glucose transporters, the GLUT family /mammalian target of rapamycin (mTOR) and insulin/IGF-1 signaling pathways  is catalyzed by the G6PD enzyme. A relatively high number of studies have interrogated the role of G6PD in different cancer types but how the overexpression of G6PD contributes to the development of different tumors is largely unknown . Yang HCgression . Anothergression . A relatgression , 86. Ghegression .fructose-1,6-bisphosphate from fructose-6-phosphate is catalyzed by PFK-1 in an irreversible step and therefore PKF-1 is considered as one of the most important rate-limiting enzyme in the process of glycolysis and is regulated by several effectors like fructose 2,6-bisphosphate , AMP and ATP. Moreover F2,6-BP levels are strongly associated with the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase  which display tissue-specific pattern. Four known isozymes of PFKFB are described  with different kinase-to-phosphatase activities  catalyzes the first substrate-level phosphorylation in glycolysis while producing 3-PG. PGK1 activity is highly responsible for maintaining energy homeostasis and serine biosynthesis. Clinically, PGK1 was overexpressed in many types of tumors. The exact molecular mechanisms for PGK1-involved drug resistance are not fully clarified but were found to be upregulated in radioresistant astrocytomas and cisplatin-resistant ovarian cancers. Moreover, inhibition of PGK1 increased the sensitivity of gastric adenocarcinoma cells to chemotherapeutic drugs 5-fluorouracil and mitomycin-C –91. Sun Proteomics profiling revealed high enolase-1 (ENO-1) expression in ER+ breast carcinomas. The high ENO-1 status was correlated with poor prognosis and with positive nodal status . Qian X The conversion of phosphoenolpyruvate to pyruvate with generation of ATP is catalyzed by PK. Till now four PK isoforms are described: PKM1, PKM2, PKR, and PKL . Apart fA large amount of literature has been published regarding LDH expression in different cancers, its effect on cancer development and progression, and its diagnostic and prognostic significance. Lactate production is an important phenomenon in the cancer microenvironment and is used as a mechanism of tumor escape from the immune response.LDHA and LDHB genes, which can assemble into five different isoenzymes as LDH1 or LDHB (H4), LDH2 (M1H3), LDH3 (M2H2), LDH4 (M3H1), and LDH5 or LDHA (M4) and is involved in the conversion of pyruvate to lactate. Deregulated levels of LDHs have been reported in multiple tumors (LDH is a tetrameric enzyme composed of two different subunits LDHA (M) and LDHB (H)  or Krebs cycle is a hub of the metabolic system. Acetyl-CoA, a common product of carbohydrate, fatty acid and amino acid metabolism enters TCA cycle that oxidizes the acetyl group of acetyl-CoA to two molecules of CO+ to NADH and CO2 release), 4. α-ketoglutarate dehydrogenase , 6. succinate dehydrogenase  and 8. malate dehydrogenase  , 2. aconitase (isomerizes citrate), 3. isocitrate dehydrogenase    related pathways in sensitive TNBC cells compared to resistant TNBC cells . Applyinin vitro and in vivo   in vivo . Another in vivo . ConsideThe increased uptake of glucose, hyperactivated glycolysis and the accumulation of lactate are the main alterations in glucose metabolism associated with different tumor types. As future challenges the following questions must be addressed and discussed in more detail:How the two frequently used ACs, DOX and epirubicin, generate distinct metabolic vulnerabilities in human breast cancer cells? This question is discussed in one recent study by McGuirk S et al. and they have found that in contrast to DOX-resistant cells, epirubicin-resistant cells present a drastic increase in OXPHOS and were more sensitive than DOX-resistant cells to treatment with phenformin. They have noticed that resistance to DOX was mostly associated with glutathione metabolism, whereas resistance to epirubicin to increased mitochondrial bioenergetic capacity . No simiDrug resistance is induced directly by one of the elements involved in the pathway  or indirectly, through an overall increase of the glycolytic metabolism or both situations can occur? Marcucci F et al. in a recent review show that both situations can be present, and the upregulation of an individual enzyme is associated with enhanced overall elevation of the glycolytic metabolism .What could be the importance of the fact that particular isoforms  of the glycolytic enzymes expressed in tumor cells are involved in the induction of drug resistance and of the fact that some of these isoforms are associated with normal cells during embryonic development? .How glucose-related metabolic changes are restored months or years after neoadjuvant therapy? Few studies address this question. According to a very recent study neoadjuvant chemotherapy worsens metabolic profile parameters  which are then recovered over 3 years. However, in the patients treated with neoadjuvant endocrine therapy there were no significant changes in fasting glucose and total cholesterol .A hot topic in the oncological treatment is how selected metabolic inhibitors work alone or how they can be used in combination as potential treatments in breast cancer cases? How metabolic inhibitors used in combination with currently used drugs result in dose reduction of the chemotherapeutic agents known to be highly toxic? In a very recent study Draguet A et al. have evaluated the effect of selected metabolic inhibitors alone and in combination in two (MDA-MB-231 and MCF-7) breast cancer cell lines. Special attention was given to the analysis of metabolic inhibitors in combination with DOX. Their results are encouraging as the combination of CB-839 (glutaminase inhibitor) and Oxamate (lactate dehydrogenase inhibitor) and the combination of CB-839/Oxamate/D609 (a phosphatidylcholine-specific phospholipase C inhibitor) resulted in remarkable cell mortality and all the inhibitors improved the efficacy of DOX in cell lines. In addition, the same inhibitors improved the efficacy of DOX. Some of the metabolic inhibitors presented in their study like AZD3965 (a selective monocarboxylate transporter 1 inhibitor) are being evaluated in preclinical trials or in clinical trials (CB-839 - a glutaminase inhibitor) .Obesity is considered as a risk factor of breast cancer especially in post-menopausal women correlating with a diminished therapeutic response and with worse disease outcome . By analElevated and dysregulated lipid metabolism is a common hallmark of cells surviving neoadjuvant therapy in breast cancer patients . StudiesWith the advent of new and more effective tools to study lipids is more and more recognized that lipids are central players in cancer biology  .It is intriguing to analyse whether the breast cancer subtype defined by the transcriptome is reflected in the lipidome of breast cancer cells or whether is there any relationship between lipidomic profile and response to different therapies. Eiriksson FF et al. by performing liquid chromatography mass spectrometry (LC-MS) to analyze the lipidome of six breast cancer cell lines of different subtypes have found differences in lipidomes within the previously defined subtypes and concluded that subtypes defined by the transcriptome are also reflected in differences in the lipidome .As it is presented in some of the recently published papers it would be also important to discuss in further reviews the utility of circulating lipid metabolites [to enhance the accuracy of known tumor markers or to distinguish tumors with early-stage vs. benign tumors ], the imBased on preclinical models lipid metabolism inhibition reversed the resistance of cancer cells to anticancer drugs suggesting that lipid metabolism play important role in drug resistance . The quePharmacological inhibitors have been developed for some of the enzymes of lipid metabolism and some of the compounds are used in combination with conventional therapies .The dysregulation of lipid metabolism may occur at different steps of the metabolic process. The most frequent aberrations associated with drug resistance are overactivation of fatty acid oxidation, elevated fatty acid biosynthesis, aberrant accumulation of lipid droplets (LD) and changes in lipid composition of cell membranes .de novo lipogenic synthesis and/or lipolytic pathway (Changes in lipid metabolism of resistant cells are considered treatment-specific and may include changes both in  pathway . Table Fatty acid β-oxidation (FAO) is a primary bioenergetic source by which fatty acids are broken down in a multistep process. Fatty acids enter cells through fatty acid transport proteins like: fatty acid translocase (FAT/CD36), tissue-specific fatty acid transport proteins (FATP), and plasma membrane-bound fatty acid-binding protein (FABPpm) , 143. InFatty acid transportation across the mitochondrial membrane is controlled by several enzymes. Once inside the cell, a CoA group is added to the fatty acid by acyl-CoA synthetase, forming long-chain acyl-CoA . The conThe transport of fatty acyl-CoAs into the mitochondria by CPT1 proteins is considered a rate-limiting step in the mitochondrial fatty acid β-oxidation pathway is . All thr2 and NADH.The β-oxidation is a cyclic process responsible for the mitochondrial disintegration of long-chain acyl-CoA to acetyl-CoA. This process is controlled by following four main enzymes: acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and ketoacyl-CoA transferase (thiolase) . In each2 is accomplished by the TCA and OXPHOS  . The thrinvasion . Targetiinvasion . Fatostainvasion .via palmitate production involving different pathways. Palmitate limits AC transmembrane uptake by cancer cells by modulating the transmembrane mobility of the drugs, a major way for AC entry inside the cells  displayed considerable side effects (e.g. anorexia). The next-generation FASN inhibitors present limited systemic toxicity in a preclinical study, higher anti-tumor potential and higher specificity for FASN , 189. TVde novo lipogenesis was observed after the inhibition of acetyl-CoA carboxylase 1 (ACC1) and 2 (ACC2) , ATP-citrate lyase (ACLY) and ACC can decrease the proliferation and growth of different cancer cells. In hepatocellular carcinoma, ND-654, an ACC inhibitor, has been shown to suppress carcinogenesis . In huma2 (ACC2) , 191. Hoasing O2 .in vivo and in vitro. Other ACLY inhibitors like difluorocitric acid and hydroxycitrate have been demonstrated to block the synthesis of FAs. Although some ACLY inhibitors performed well and have been validated, most of the studies to date are still in the preclinical phase  enzyme as another rate-limiting enzyme in fatty acid synthesis that converts saturated acids to monounsaturated fatty acids (MUFAs) involved in many biological processes . Accordingly more and more studies highlight different roles of SCD1 like modulation of autophagy  others dvia passive diffusion or endocytosis  also playing an important role in cancer growth and/or oncogenic signaling pathways it is reasonable to consider the significant role played by cholesterol in tumor metabolism. Cholesterol deficiency in cell membranes has been shown to inhibit cancer progression  as well as considering its role in forming lipid rafts, an indispensable cell membrane structure in cancer cells. As many tumor-related proteins are located in lipid rafts this cell component is an important platform for oncogenic signaling pathways. Considering other aspects like the metabolites obtained by gression  and cholgression , 200.via LDL receptor (LDLR)-mediated endocytosis. In the mevalonate pathway 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) is reduced to mevalonate by HMG-CoA-reductase enzyme considered as the primary control site for cholesterol biosynthesis  by inducing changes in the organization of lipid rafts are preferred .The accumulation of lipid droplets is a less well-studied aspect of chemoresistant cancer cell lines . Lipid dSirois I et al. in a very impressive study by analyzing a novel MDA-MB-436 cell-based model of chemoresistance characterized by a unique and complex morphologic phenotype with numerous lipid droplets and a set of primary chemoresistant TNBCs have identified several metabolic vulnerabilities. These include a dependence on perilipin family member perilipin 4 (PLIN4), a protein coating of the observed lipid droplets that play important role in fat mobilisation, expressed both in experimental conditions (in the chemoresistant TNBC cells) and in chemoresistant tumors treated with neoadjuvant DOX-based chemotherapy. Their results call attention to a novel mechanism of chemotherapy resistance that may have therapeutic consequences in the treatment of drug-resistant cancer .Based on the above-mentioned data related to lipid metabolism in breast carcinomas there are some intriguing questions to be analyzed in more detail:de novo fatty acid synthesis and oxidation as sources for energy requirements the TNBCs are characterized by overexpression of the genes involved in the utilization of exogenous fatty acids. They suggest that treatments have to be tailored to individual subtypes  subtypes depend on a balance between subtypes .Lipid metabolism has been difficult to analyze partly due to technical problems. Considering the challenges  it is questionable how the new technologies could help in delineating the contribution of lipid metabolism to tumor differentiation, progression and resistance to different drugs. Matsushita Y et al. summarize the new technology currently used in lipidomics and confirm that recent innovations especially in mass spectrometry- and chromatography-based lipidomics technologies have improved our understanding of the role of lipids in cancer .Cancer-associated adipocytes are poorly investigated cells in breast tumor microenvironment. What should be the role of the surrounding and breast cancer-associated adipocytes? A few studies suggest that cancer-associated adipocytes can also cause resistance to radiotherapy and to chemotherapeutic drugs , 210. AnHow is lipid profile of patients restored after (neo)adjuvant chemotherapy? Tian W et al. suggested that different age groups showed different changes in lipid levels and in general the lipid profiles were restored to baseline levels approximately 6 months after chemotherapy completion .Main metabolic alterations in breast cancers include the preference of the glycolytic pathway , the enhanced oxidative phosphorylation pathway and dysregulation of fatty acid metabolism.Current studies present the important aspect of tumor metabolism associated with anticancer drug resistance and the association of the partly unknown aspect of the metabolic plasticity in chemotherapy resistance. Efforts are made to combine chemotherapies with drugs targeting different steps of metabolic pathways. Compared to a huge amount of results related to the above-mentioned questions only a few studies present data related to changes in metabolic profile induced by currently applied chemotherapeutic agents used in neoadjuvant or adjuvant settings. There are also gaps in analysing the association of the metabolic changes induced by chemotherapies with different response rates to the applied treatment regimen or with tumor progression.Considering just some of the points like: the regulation of lipid and glucose metabolism is also important for normal cells, the intra- and intertumoral metabolic heterogeneity in breast cancers, the different metabolic signatures observed in different breast carcinoma subtypes, the lack or imperfect technologies to study components of metabolic pathways, it is still a huge challenge to find substances that target different steps of glucose and lipid metabolism in the very heterogeneous group of breast tumors without affecting normal cells. Additionally, current metabolomic and especially lipidomic approaches have identified several partly unknown components of metabolic pathways paving the way to a better understanding of cancer biology. As exemplified in our and in recently published reviews there are relatively broad opportunities in targeting metabolism in cancers but it is a key question whether we have considered nearly all metabolic hubs as targets and whether we can rewire our mind to optimise the therapies based on the several new data.AMT: conceptualization, interpretation of data, writing original draft, review. JK: review, interpretation of data. SV-K: editing, preparing figures. BT: review, editing, preparing figures and supervision. All authors contributed to the article and approved the submitted version.This study was supported by the CELSA 19/037 grant." FAT-MetBC: Facing Increased Adiposity in Treating Metastatic Breast Cancer.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Thulium laser (Tm:YAG) prostate surgery is a safe and effective procedure with low morbidity and comparable clinical outcomes to those of transurethral resection of the prostate (TURP). However, the sexual function outcomes (erectile and ejaculatory function) have been scarcely studied.We aimed to assess the impact of Tm:YAG prostate surgery on sexual outcomes (erectile and ejaculatory function) and compare them with those patients undergoing TURP.We searched digital databases like PUBMED, SCOPUS, CENTRAL and EMBASE using relevant keywords to identify comparative studies on TURP and non-comparative studies on Tm:YAG prostate surgery that assessed sexual outcomes. We performed qualitative and quantitative analyses with the extracted data. We carried out a meta-analysis to compare postoperative International Index of Erectile Function (IIEF-5) scores and incidences of retrograde ejaculation (RE) in patients undergoing either Tm:YAG or TURP. The pre-operative and post-operative IIEF-5 scores were pooled to estimate overall scores.P = .001). We found no significant associations between the procedures. The pooled OR for the association of RE was estimated at 0.90 .We included 5 comparative and 8 non-comparative studies in this review. We found the postoperative IIEF-5 score improvements to be significantly higher in the Tm:YAG prostate surgery group than in the TURP group with a significant mean difference (MD) of 0.45 (95% CI, 0.18 to 0.72; Tm:YAG prostate surgery improves erectile function more than TURP, according to our findings. Tm:YAG prostate aided surgery also outperforms TURP in terms of preserving sexual function following surgery.However, We found similar or no difference in incidence of RE between Tm:YAG prostate surgery and TURP.Bibo L, Hao L, Pang K, et al. Assessment of Sexual Outcomes in Patients Undergoing Thulium Laser Prostate Surgery for Management of Benign Prostate Hyperplasia: A Systematic Review and Meta-analysis. Sex Med 2022;10:100483. Benign prostatic hyperplasia (BPH) causes lower urinary tract symptoms in most elderly men.5,Transurethral resection of prostate (TURP) is the gold standard surgical treatment; during the procedure the prostate lobes are resected with a trans-urethral resectoscope without any incisions.,,The safety and efficacy of Tm:YAG laser was assessed in a recent systematic review,Another systematic reviewRegardless of the procedure, prostate resections are associated with urinary and sexual complications.Another recent systematic reviewHowever, no meta-analysis has evaluated the impact of Tm: YAG prostate surgery on sexual outcomes. Thus, we systematically searched the literature for all studies assessing the impact of Tm:YAG prostate surgery on sexual outcomes (erectile and ejaculatory function) and compared and pooled the data with those available from studies on TURP.We carried out the systematic review and meta-analysis under strict accordance with the Preferred Reporting of Items for Systematic reviews and Meta-Analysis (PRISMA) guidelines.How does Tm:YAG prostate surgery compare to TURP for BPH management in terms of sexual outcomes?What are the sexual outcomes after Tm:YAG prostate surgery for BPH management?th February 2021 from the starting year of publications. We also performed a manual search in the following relevant peer reviewed indexed journals: World journal of urology, BMC surgery, Asian Journal of Urology, Prostatic diseases, Prostate International, and The Prostate. In addition, we thoroughly screened the bibliography section of relevant systematic reviews for any possible eligible studies. Finally, we also searched for any ongoing or unpublished trials on trial registries and other grey literature databases.We searched electronic databases like PUBMED, SCOPUS, CENTRAL and EMBASE using the following search string: (benign prostate hyperplasia) OR (prostate) OR (prostate surgery) OR (prostate enlargement) AND (TRUP) OR  OR (Thulium laser) OR (Tm:YAG) OR (Vapo-enucleation). We modified and adapted the search keywords to fit in respective databases. The identified relevant keywords were combined using Boolean operators ‘AND’, ‘OR’ & ‘NOT’. We conducted the last electronic search for all databases in February 2021. The search included the publications till 20We imported the retrieved reports into a citation manager  to automatically discard duplicates after the search on multiple databases. 2 independent reviewers  screened titles and abstracts of retrieved studies to identify eligible articles for full text assessment. The screening was carried out by to avoid chances of missing out any relevant reports. We selected the final full texts of the eligible articles based on a predefined inclusion and exclusion criteria.The following PICO selection criteria for eligible publications with population focussed to elderly men with BPH undergoing Tm:YAG prostate surgery as intervention compared to TURP assessing sexual outcomes like change in IIEF-5 scores and ejaculation function).vs. TRUP) reporting sexual outcomes.All single arm interventional (Tm:YAG prostate surgery) and comparative studies , involving modified approaches for prostate resection and Studies published in a language other than English were excluded.We extracted the data from the selected studies onto the Microsoft Excel spreadsheets for better visualization and convenience. We segregated the relevant information from both comparative and non-comparative studies into 2 different spreadsheets. We recorded characteristics of study , demographics , interventions , and sexual outcomes (IIEF-5 scores and incidence of RE and hemospermia) for each study. We also obtained pre-operative and post-operative scores and IIEF-5 changes wherever possible. We contacted authors of articles with missing or unclear information by email for clarification.2 statistics and considering heterogeneity as low for I2 values < 40%, moderate for values ranging between 40 and 70%, and high for values > 70%.We subjected the data to both qualitative and quantitative analyses. We tabulated the study, demographic, and interventional characteristics for both comparative and non comparative prospective or retrospective studies. We performed qualitative analyses on outcomes that could not be pooled quantitatively. We used the quantitative data for the meta-analysis. We expressed continuous outcomes (like IIEF-5 scores) as means and standard deviations and calculated IIEF-5 score changes to compare the impact on erectile function after either TmLRP or TURP. We used dichotomous data (like RE) to calculate odd's ratios (ORs) and compared the values obtained for each procedure. We pooled pre-operative and post-operative IIEF-5 scores obtained from the single-arm non-comparative studies to get overall scores. The comparative meta-analysis was carried out using the RevMan 5.4 software. We used the Open Meta-Analyst software for the single arms pooling of means and mean difference scores. All the meta-analyses were carried out using a random-effects model, considering the cohort variations in the included studies. We assessed the heterogeneity among the included studies using IWe used the risk of bias assessment tools to evaluate all included studies. 2 independent reviewers  conducted a risk of bias analysis for RCTs using the Cochrane risk of bias tool. The trials were analyzed for bias in selection of participants by evaluating the randomization process and allocation concealment methods; bias in blinding of participants and personnel; bias in blinding of outcome assessors; bias in selective reporting of results, and lost to follow-up. The risk of bias in each study was graded as low, moderate, or high based on the adequacy of the above-mentioned domains.Two independent reviewers  used the ROBINS-I (Risk of Bias in Non-randomized Studies - of Intervention) tool to assess the risk of bias of non-randomized studies; and the discrepancies were resolved by discussion and consensus with a third reviewer (JZ). We graded 7 bias domains for each study as high, unclear, or low risk: bias due to confounding, bias in selection of study participants, bias in exposure classification, bias due to departures from intended exposures, bias due to missing data, bias in measurement of outcomes, and bias in selection of reported result. Based on these domains, we categorized the studies as having low risk of bias , high risk of bias (if one or more domains were at high risk); or medium risk of bias (if 2 or more domains had unclear risks).A pool of 1687 articles was retrieved from various digital and manual sources. After title and abstract screening, we deemed only 25 reports as eligible for full text assessment. We found proper reasons for excluding thirteen articlesThis systematic review and meta-analysis included a total of thirteen studies,,,,,,,,Table. 2Four RCTs,,,We included 8 prospective cohorts38https://training.cochrane.org/resource/revman-calculator).We included data from eleven studies in the meta-analysis. The data presented in median and inter-quartile range was converted to mean and standard deviation using the excel tool provided by Cochrane Collaboration . The heterogeneity between the studies was 0%. Please see Only 2 studies,We analyzed data from 7 non comparative prospective cohort studies39,38,,We also carried out a subgroup analysis to assess the change in IIEF-5 score based on the technique employed. We pooled data from 5 studies,,P = .71; I2 = 0%). See We also compared the number of RE events postoperatively between the Tm:YAG prostate surgery and TURP groups.,,,,,,,We found the risk of bias for non-randomized studies to be moderate for 5 studies,,,We found the risk of bias for randomized clinical trials to be high for 3 studiesWe designed this systematic review and meta-analysis to compare sexual outcomes (erectile and ejaculatory) after either Tm:YAG prostate surgery or TURP for BPH. We included data from 5 comparative studies and 8 single arm prospective cohorts assess erectile and ejaculation function after the treatments. We focused on the IIEF-5 scoring to evaluate erectile function both pre-operatively and postoperatively in all included studies. The IIEF-5 is a multidimensional validated questionnaire comprising 5 domains of sexual function .We found the IIEF-5 score improvement to be significantly higher in patients undergoing Tm:YAG assisted prostate surgery than in those undergoing TURP. It is generally believed that The heating effect of the electrocautery in TURP is thought to cause cavernous nerve and vascular injuries, and the neuropraxia and emotional stress may lead to impotence after TURP. We also found that the IIEF-5 scores improved over time . The improvements at the 6- and 12-month follow-ups were comparatively higher than that at the 3-month follow-up. Similar results were also found in the systematic review by Cornu et al. 2015The erectile function of patients undergoing vapo-enucleation with thulium laser was better post-operatively than that of patients undergoing Thulium laser-assisted enucleation and resection. TmLEP is a similar procedure to TmVEP except for the fact that the enucleation is done mechanically without the use of energy in TmLEP. However, the initially incisions are made by the laser. TmVEP uses continuous pulses of thulium laser to vaporize the prostate lobes. The mechanical removal of the prostate lobes always threatens to cause cavernous nerve injury, and this may explain the apparent superiority of TmVEP. Hood sparing techniques at the level of the apex (36) are now-a-days in practice could also be a reason of the same.RE was the only ejaculatory dysfunction (ED) recorded in 3 RCTs. RE occurs when semen enters the bladder instead of emerging through the penis during orgasms. Although the patients reach sexual climax, they ejaculate  little or no semen. RE can have a deleterious effect on the quality of life by reducing orgasmic intensity and inducing anxiety and depression.The pathophysiology of treatment-related RE remains unclear, but the predominant contributing factor seems to be the resection of the bladder neck and proximal prostate portions, which disrupts the internal urethral sphincter that maintains the antegrade ejaculatory function.To our knowledge, this is the first review assessing sexual outcomes after thulium laser-assisted prostate surgery and comparing them to those after TURP. A systematic comparing the sexual function outcomes between endoscopic enucleation and TURP included only oneWe were able to conduct subgroup analyses based on the follow-up durations and surgical techniques, which painted a clear picture. However, our review also has limitations. First, uni-polar and bi-polar TURP were both considered as the TURP gold standard for the control groups. Second, we found high bias risks in the included RCTs and this could reduce the credibility of our results. Third, we failed conduct subgroup analyses based on the 5 IIEF-5 scoring criteria domains because only 1 study, out of all included studies, presented the scores for all domains. Finally, the heterogeneity among the studies was high; this may be due to inclusion of different grades of patients presenting mild to severe BPH. The limited availability of comparative studies could not provide a clear comparison between TURP and TmLEP and TmVEP separately. Further evidences are necessary to answer the differences better.Within the limitations of this review, Tm:YAG prostate surgery showed better change in IIEF-5 scores post-operatively and resulted in better erectile function outcomes than TURP. Moreover, the IIEF-5 score improvements were higher after the vapo-enucleation of prostate procedure than after simple enucleation using Thulium laser. We found an association between RE events and both Tm:YAG and TURP without significant differences. Although the findings were based on statistical analysis, the clinical benefit may still be unclear. Further clinical trials with rigorous methodology and larger sample sizes are needed to strengthen the evidence.Not applicable.


In order to support primary care during the first pandemic wave (March 2020), the Italian Government instituted multiprofessional health teams called “USCA” , which ensured continuity of care for COVID-19 patients who do not need hospitalization. The aim of our study was to compare the volumes of USCA service utilization in Florence  during the peak of home visits of three pandemic waves.This single-center study followed a retrospective cross-sectional design. The USCA of the Heath District of Florence served a population of 366,190 people. The following data were collected: home medical visits, nursing home (NH) visits, visits in health-care hotels. The peak periods of three epidemic waves were considered in the analyses: the second wave (23 October - 20 November 2020), the third wave (25 March - 22 April 2021), and the Omicron period (27 December 2021 - 6 February 2022). The maximum 7-day moving averages of the daily number of visits during the three periods were calculated. Relative percent differences for visits comparing the considered periods were computed.Home visits during the third pandemic wave increased by 14% compared to the second wave , while a decrease was observed during the Omicron period . Visits in health-care hotels during the third wave doubled compared to the second wave. After the start of the COVID-19 vaccination campaign, NH visits steeply declined . During the Omicron period, NH visits increased by almost four times compared to the third wave period.The USCA service utilization was significant in all the analyzed periods. In a pandemic context, it is necessary to strengthen primary care services such as USCA, which have proved to respond to rapidly changing health needs.• The USCA service is an innovative model of integrated home care that has proved to respond to rapidly changing health needs during all phases of the COVID-19 pandemic.• The USCA service utilization was significant during all phases of the pandemic. The USCA service has introduced new ways of working and new relationships between services in primary care.


While decreased alpha-band functional connectivity (FC) and changes in network topology have been reported in Alzheimer’s disease, it is not yet entirely known whether these differences mark cognitive decline in the early stages of the disease.Our study aimed to analyze EEG FC and network differences in the alpha frequency band during visuospatial memory maintenance between Mild Cognitive Impairment (MCI) patients and healthy elderly with subjective memory complaints.FC and network structure of 17 MCI patients and 20 control participants were studied with 128-channel EEG during a visuospatial memory task. FC was measured by amplitude envelope correlation with leakage correction (AEC-c), while network analysis was performed by applying the Minimum Spanning Tree approach.Increasing memory load enhanced the mean alpha-band FC in the control group. In contrast to that, after an initial increase, the MCI group showed significantly (p<0.05) diminished FC in the highest memory load condition. Mean alpha AEC-c correlated significantly with the size and mean diffusivity of medial temporal lobe structures in the entire sample. The network analysis revealed a rerouted network in the MCI group with a more centralized topology and a more unequal traffic load distribution compared to the control group.Alpha-band FC correlates with cognitive load-related modulation, with medial temporal lobe atrophy, and with the disruption of hippocampal fiber integrity in the earliest stages of cognitive decline. The more integrated network topology of the MCI group is in line with the “hub overload and failure” framework and might be part of a compensatory mechanism.No significant relationships.


Obsessive-compulsive disorder (OCD) is a disabling psychiatric disorder that affects 2-3% of the population. Pharmacological or cognitive behavioral therapy can reduce symptoms. Deep brain stimulation is emerging for treatment-resistant patients.We measured neuronal activity in two patients with treatment-resistant OCD, who had DBS electrodes implanted bilaterally in the BNST. Local field potentials were recorded directly from the BNST during and without symptom provocation and without electrical stimulation.In two patients with a diagnosis of treatment resistant OCD (TR-OCD) local field potentials (LFP) were recorded as part of their clinical follow up post-implantation. In both patients, the diagnosis of TR-OCD was confirmed by a neuropsychiatric examination and a multidisciplinary committee comprising both experienced psychiatrists and neurosurgeons from different medical centers in Belgium. We used BrainSense recording technology in the Percept PC to record the LFPs. The LFP recordings in the first patient were acquired on the 15th day after DBS surgery. In the second patient, the interval between implantation and recording was 18 days. Symptom provocation was performed using the MOCCS image set, developed by Mataix-Cols.At rest, relative power peaks in the BNST were highest in the theta (4-8 Hz) frequency band for both patients. In both patients switching DBS ON during provocation images appears to cause the LFP signal to closely resemble that recorded during neutral images.The main finding of this pilot study is that switching stimulation ON in the BNST during provocation images causes the LFP signal to more closely resemble the LFP recorded during neutral images.No significant relationships.


Herpes zoster is caused by varicella zoster virus. After the previous attack of chickenpox, it remains dormant in body and reactivation causes herpes zoster or shingles. There is prodromal of pain for 1 -4 days. Erythema and oedema are followed by formation of groups of vesicles. The crust form in 15 days without scars or minimal scars. Thoracic intercostal nerves and ophthalmic division of trigeminal nerve most frequently affected. Other spinal nerves also involved. A 64-year-old lady come to Outpatient Department (OPD) with vesicles on waist and right thigh with complaint of burning and severe pain in lumbar area. The patient had taken pain killers/anti-inflammatory drugs over the counter but of no use.


Combinations of two different sintering schedules (direct-sintering and step-sintering) and three different heating rates  were applied to study the densification, shrinkage, and microstructural evolution of BJAM parts. This study demonstrated that, although the green density of the BJAM samples was ∼42% of the theoretical density, the sintered parts experienced large linear shrinkage up to ∼25% and reached ∼97% density without compromising shape fidelity. This was ascribed to a more homogeneous pore distribution throughout the part before ramping up to the SLPS region. The synergistic effects of carbon residue, the slow heating rate, and the additional isothermal holding stage at the solid-phase sintering region were determined to be the key factors for sintering BJAM WA low-alloy steel powders with resulting minimal entrapped porosity and good shape fidelity.This work investigates the feasibility of net shape manufacturing of parts using water-atomized (WA) low-alloy steel with comparable densities to conventional powder metallurgy parts via binder jetting additive manufacturing (BJAM) and supersolidus liquid phase sintering (SLPS). In this study, a modified water-atomized powder grade with similar composition as MPIF FL-4405 was printed and pressure-less sintered under a 95% N  The bin2 or CO atmosphere) and/or carbon source  to reduce surface oxides and prevent their transformation into more thermodynamically stable ones during sintering Owing to the low green density (42%), the densification up to 97.1% corresponded to the linear shrinkage of ∼25% on the z-axis, and ∼23% on the x/y-axis. Nevertheless, the surprisingly large shrinkages have a negligible effect on the geometrical precision of parts for the geometries used in this study due to an improved pore morphology using optimized sintering conditions.This study examines the ability to sinter WA low-alloy steel fabricated by BJAM process to a relative density of 97.1% via supersolidus liquid phase sintering (SLPS) with minimal shape loss. A comprehensive investigation was completed to identify any potential challenges in the densification of oxygen-containing steel powders with a significantly lower green density compared to traditional PM green parts, which are typically around 80%. The following major conclusions are drawn from the present study.In the future, a systematic investigation of the processing window of BJAM WA low-alloy steel parts with larger dimensions and more complex geometry will be undertaken.Mingzhang Yang: Conceived and designed the experiments; Performed the experiments; Analyzed and interpreted the data; Wrote the paper.Mohsen K. Keshavarz: Conceived and designed the experiments; Wrote the paper.Mihaela Vlasea: Conceived and designed the experiments.Amin Molavi-Kakhki and Martin Laher: Conceived and designed the experiments; Contributed reagents, materials, analysis tools or data.10.13039/501100006266Economic Development Agency for Southern Ontario, Canada [814654].Assistant Prof. Mihaela Vlasea was supported by Federal Data will be made available on request.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Second, secondary membrane growth was applied to give rise to a continuous and homogeneous crystalline MOF membrane layer. The gas permeances  tests confirmed high membrane permeability with no macro-defects. The as-prepared membranes that were used for dye separation (Rhodamine B) showed relatively good separation capacity.Metal Organic Frameworks (MOFs) present high surface areas, various pore topology as well as good stabilities. The functionalities and porosity can be tuned by using different linkers with various functional groups and a wide range of linker lengths. These properties make them good candidates in membrane separation applications. In this work, we propose a simple UiO-66 MOF-based membrane fabrication method following two steps. First, the α-alumina tubular membrane support was dip-coated with MOF-polymer hybrid nanoparticles (NPs). These NPs were prepared via one-pot synthesis by adding poly (methacrylic acid)- Metal Organic Frameworks (MOFs) are well-known crystalline porous materials based on metal ions or clusters coordinated to organic linkers following the principle of reticular chemistry ,2. MOFs 2 membrane. Drobek et al. . B. B2 , or the small quantity of UiO-66 or UiO-66-NH2 present on the membrane support (only few micrometers thick layer of deposited MOF). Moreover, the α-alumina support showed intense diffraction signals that could mask the UiO signals.The experimental PXRD patterns  confirme2, CO2 and SF6) were performed at different transmembrane pressures (ΔP) from 0.25 up to 1 bar. Such a series of measurements allowed the mapping of the evolution of the gas permeability versus the transmembrane pressure and the determination of the ideal selectivity of the selected gas pairs.The as-prepared MOF membranes were characterized by single gas permeation measurements in order to check the quality of the membrane layers and their performance (membrane compaction and gas selectivity). Four pure gases .In order to evaluate the membrane separation properties, ideal selectivity for selected gas pairs were calculated . When ev2 secondary growth with the best gas selectivity results obtained from the single gas permeation measurements was used for the removal of RhB from an aqueous solution. The membranes were conditioned at a higher pressure as compared to the filtration working pressure prior to dye filtration. This step ensures that the membrane thickness would be compressed to a stable level. As can be seen in −2 h−1 bar−1  and then was soaked in 5 mL of ethanol for 24 h. It was observed that the pink color was removed from the membrane and the water/ethanol had become colored; thus, indicating that the removed dye could be washed off from the membrane. Potentially, both the membrane and the separated dye could be recycled. However, this requires further studies.The membrane prepared with UiO-66-NH−1 bar−1 . The dec2 membranes via a direct seeding of UiO-polymer hybrid colloidally stable NPs followed by the secondary growth of UiO-66 or UiO-66-NH2. Unlike other reported reactive seeding or polymer-assisted seeding methods, there was no need for the precursor immobilization or any extra steps to mix the MOF crystals with the polymer. Thus, the synthesized UiO-polymer hybrid NPs were stable in a colloidal solution and could be easily deposited on the support via dip-coating, resulting in the formation of a thin and homogenous seed layer.In this work, a simple and innovative procedure is reported for the preparation of UiO-66 and UiO-66-NH2 crystals. The gas permeance tests confirmed a high membrane permeability but low selectivity towards the four tested gases  suggesting the presence of some inter-crystalline defects in the membrane structure. Considering the membrane pore size, it is expected to be an attractive candidate in the fields of liquid separation (water or organic media). The RhB filtration corroborated the membrane stability and showed that the membrane has a good dye selectivity with a relatively high permeability. The membrane quality needs to be further improved by optimizing the secondary growth protocol to reach the level of molecular sieving. The data reported here, along with our previously reported work [This direct seeding of MOF-polymer colloidally stable NPs could be extended to other carboxylic acid-based MOFs for seeding. The new reported method allows facile seeding on different types of supports , potentially finding direct application in industry since they would be able to replace conventional filtration systems. The as-seeded supports were then subjected to classical solvothermal secondary MOF growth method, leading to UiO-based crystalline and compact membranes with interconnected UiO-66 and UiO-66-NHted work , highlig


Amidst an ever-evolving pandemic, the demand for timely and accurate diagnosis of coronavirus disease 2019 (COVID-19) continues to increase. Critically, managing and containing the spread of the disease requires expedient testing of infected individuals. Presently, the gold standard for the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains the polymerase chain reaction (PCR) test. Potential vulnerabilities of this testing methodology can range from preanalytical variables to laboratory-related analytical factors and, ultimately, to the interpretation of results. The coronavirus disease 2019 (COVID-19) pandemic represents a rapidly evolving public health emergency. The emergence of viral variants, a lack of diagnostic equipment and expertise coupled with inequitable vaccine access have resulted in global outbreaks. Consequently, the demand for testing continues to increase. The gold standard for the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains the polymerase chain reaction (PCR) test. This test, however, requires skilled staff, laboratory infrastructure and specialised equipment. Moreover, there are numerous factors that influence the outcome and interpretation of a SARS-CoV-2 PCR test. This article will highlight potential pitfalls to consider when using this testing methodology.1 The most common method used for respiratory specimen collection is the nasopharyngeal swab (NPS). Another specimen that may be collected is the oropharyngeal swab; however, data suggests that it is slightly less sensitive than the NPS.4 Saliva has been evaluated in various studies globally as it is a simple and non-invasive sample type that may be taken by a patient at home, thereby limiting contact with healthcare workers and the need for costly personal protective equipment. Moreover, saliva does not require expensive collection supplies and appears stable under variable conditions for a prolonged period.5 A recent systematic review of the diagnostic performance of different sampling approaches for SARS-CoV-2 reverse transcriptase polymerase chain reaction (RT-PCR) testing found that combined nasal and throat swabs gave the highest sensitivity of 97%. Lower sensitivities were obtained from saliva specimens (85%) and nasal swabs (86%) with the lowest sensitivity achieved from throat swabs (68%).6The molecular diagnosis of COVID-19 rests on the detection of viral ribonucleic acid (RNA) in a clinical specimen. Infection begins in the upper respiratory tract (URT) and may progress to the lower respiratory tract (LRT) in more severe cases.8 However, both these sample types are not routinely used in South Africa (SA) for PCR testing.The Centre for Disease Control (CDC) and the South African National Department of Health have also recently approved mid-turbinate and nasal swabs for testing.9; however, shortage of collection supplies has led to the use of cotton swabs, especially in poorly resourced healthcare sectors. Whilst a recent study found that cotton does not inhibit the detection of SARS-CoV-2 RNA, further studies are required to confidently confirm this.10Suboptimal sample collection influences the amount of RNA detected. Specimen collection devices should have aluminium or plastic shafts as wooden shafts may contain substances that inactivate some viruses and inhibit PCR testing. The World Health Organization (WHO) recommends synthetic-tipped swabs such as rayon or nylon for SARS-CoV-2 nucleic acid detection11Operator technique is another critical factor that has often contributed to false-negative results. Improper technique results in swabs not reaching the target site; the aim is to brush infected cells and secretions off the nasopharyngeal mucous membranes. This ensures that sufficient RNA is obtained from the site of initial viral replication. There has been an influx in the number of drive-through sites at which tests are being performed to improve convenience and increase the number of tests done whilst adhering to social distancing recommendations. Despite the advantages offered by a drive-through model, literature interrogating the accuracy on SARS-CoV-2 PCR results is scant, as patient and operator positioning may not be optimised for proper NPS technique.12 Lower respiratory tract specimens include expectorated sputum, bronchoalveolar lavage (BAL) or endotracheal aspirates and should be submitted in clean universal containers without transport medium. These samples are especially useful when the pretest probability of disease is high and a URT specimen yielded a negative PCR result. A study evaluating the detection of SARS-CoV-2 RNA in a range of sample types found that BAL specimens showed the highest positivity rates (93%) followed by sputum (72%).13Sample type may also affect a PCR result. Patients with evidence of LRT disease may require an LRT sample, especially if the patient is tested later in the course of illness.14 Ideally, samples should reach the laboratory as soon as possible and arrive in viral or universal transport medium at 2 °C – 8 °C. Dry swabs in a sterile tube may also be submitted. Dry swabs can be sent at ambient temperature but should reach the laboratory within 48 h. Unfortunately, testing delays and backlogs have led to widespread fluctuations in transport and storage conditions. A United States-based study examining the effect of extended storage at ambient temperature on NPS found minimal and clinically insignificant impact on PCR results.15 Research is ongoing for molecular testing in lower-income settings under various storage conditions. In the interim, it is crucial to abide by the recommended storage and transport conditions to maintain sample integrity.Transport and storage of samples may also affect the integrity of viral RNA.15 Therefore, an NPS taken too early or too late in relation to clinical symptoms may cause a false negative result. Other pre-analytical errors such as sample swaps, mislabelling and contamination may also contribute to false PCR results.Timing of testing in relation to disease course is critical when submitting a sample to the laboratory. Viral load in the nasopharynx is highest in the immediate presymptomatic and symptomatic phase of the illness and decreases from week 3 to eventually become undetectable.16A SARS-CoV-2 PCR test is a very sensitive molecular method of reverse transcribing SARS-CoV-2 RNA into DNA, followed by amplification of the target genomic sequence. Fluorogenic probes are used to search for the target DNA sequence which generates a fluorescent signal that increases proportionally to the amount of viral RNA present in the sample. A cycle threshold (Ct) value is obtained when the fluorescence reaches a specific threshold within a certain number of PCR cycles. A Ct value less than 40 for one or more viral gene segments is generally reported as PCR positive. There is generally an inverse correlation between Ct values and viral loads, such that lower Ct values represent higher viral RNA loads.env), nucleocapsid (N), spike (S), RNA-dependent RNA polymerase (RdRp) or ORF1 genes.17 For any sample, these targets should take roughly the same number of PCR cycles to be detected. The Foundation for Innovative New diagnostics (FIND) is compiling a database of SARS-CoV-2 molecular assays that are commercially available or in development for the diagnosis of COVID-19, accessible at https://www.finddx.org/covid-19/pipeline/?section=molecular-assays#diag_tab. Currently, within the National Health Laboratory Service in SA, the six most widely used testing kits are presented in Specific SARS-CoV-2 gene segments are targeted by different assays. Generally, most PCR assays target two or three of the envelope , the distance between the sample collection site and the testing laboratory, the volume of tests received at the laboratory and the assay(s) used.18 Cross-contamination from a positive sample to a negative one may occur during sample preparation for PCR. This risk increases substantially during peaks of infection when viral loads in samples are very high. During the PCR test, billions of copies of nucleic acid are generated, and these can contaminate instruments, reagents and samples. A lesser source of contamination is assay-derived, which can cause false positives and reduce the specificity of the diagnostic assay.19 Whilst quality assurance steps such as the use of extraction controls and negative template controls may help to detect contamination, low-level contamination may go unnoticed. This scenario can occur during a peak of infection when there are many samples containing millions of copies of viral RNA, coupled with an urgency in the laboratory to ensure that samples are processed rapidly.Contamination has been a major thorn in the side of molecular assays and may be derived from two sources: cross-contamination between specimens or synthetically derived target nucleic acid.20 As the pandemic has progressed, several genetically unique variants of concern, monitored by the WHO, have emerged globally, namely, B.1.1.7 (Alpha), B.1.351 (Beta), B.1.617.2 (Delta) and P.1 (Gamma).21 Because molecular tests detect a specific viral nucleic acid sequence, a mutation may affect the performance of a PCR test if it occurs in a region of the genome targeted by the test. The impact of these variants on test performance is influenced by the design of the test, the sequence of the variant and the prevalence of the variant in the population. For example, the alpha variant carries a double deletion at positions 69 and 70 on the spike protein gene (S-gene). This appears to impact detection using an assay that targets the S-gene, such as the TaqPathTM COVID-19 Combo Kit.22 This S-gene target failure was also observed with the recent Omicron variant which contained the same mutation. Fortunately, even if a mutation impacts one of the PCR targets, most commercial PCR-based tests have two or more targets to detect SARS-CoV-2. In the case of a mutation, one may consider retesting using a test targeting different genes if COVID-19 is still suspected after receiving a negative PCR test result.The SARS-CoV-2 has a high rate of error-prone replication and penchant for recombination.23 This may be a result of a sample taken very early in the infection, intermittent viral shedding – usually at the tail-end of infection – or contamination.If the PCR assay detects all viral gene targets , then viral RNA is detected and the test is positive; if no targets are detected, then viral RNA is absent from the sample and it is reported as a negative result; if only some of the targets are present, then the laboratory takes a decision based on an agreed Ct cut-off value whether to report this result as positive or as inconclusive. An inconclusive result can be the most problematic to troubleshoot. Polymerase chain reaction-based assays can yield a weak signal or nonspecific result near the limit of detection of the assay. This high Ct value generally indicates very little viral RNA in the sample.26 If one correlates Ct values with clinical progression, Ct values are lowest in the immediate presymptomatic and first week of the symptomatic phases of COVID-19.27 Therefore, Ct values correlate closely with a clinical history and examination. Unfortunately, these details are often omitted on a request form, so that interpretation of a result is less case-specific. Moreover, PCR can often detect viral RNA for weeks after the resolution of symptoms.28It is important to remember that PCR is an extremely sensitive method and under optimal conditions can detect fewer than 10 copies of viral RNA in a clinical sample; however, it cannot distinguish between viable virus and noninfectious RNA. Virus viability appears to be brief with viral culture negative in cases with a Ct higher than 33.29 Whilst Ct values from patient samples have demonstrated clinical utility in certain scenarios,30 including Ct values routinely on a PCR result report could be misleading to the clinician.31 It is thus critical for the requesting healthcare worker to discuss the PCR result with the clinical virologist whilst considering a patient’s history, contacts, symptoms, radiological features and previous laboratory results. From a laboratory perspective, thorough knowledge of the performance characteristics of individual PCR assays in use is necessary to accurately interpret results.32 Furthermore, a sample tested on different assays can yield different Ct values reflecting differences in the targets detected and the chemistry of the test used. Consequently, when considering trends in Ct values it is preferable to test samples with the same assay or platform each time.Issuing binary results may create confusion by equating a sample with a very high viral load to one with a significantly lower viral load.The COVID-19 pandemic has prompted a rapid upscaling of diagnostic platforms in both private and public laboratories. Whilst the PCR test currently remains the gold standard for the diagnosis of COVID-19, it is not without challenges. Pre-analytical, analytical and postanalytical factors may influence the outcome of a result. Alternative diagnostic algorithms incorporating immunoassays and novel molecular methods are currently under investigation.


Lung cancer is currently the leading cause of cancer-related deaths worldwide. Significant improvements in lung cancer therapeutics have relied on a better understanding of lung cancer immunity and the development of novel immunotherapies, as best exemplified by the introduction of PD-1/PD-L1-based therapies. However, this improvement is limited to lung cancer patients who respond to anti-PD-1 immunotherapy. Further improvements in immunotherapy may benefit from a better understanding of innate immune response mechanisms in the lung. Toll-like receptors (TLRs) are a key component of the innate immune response and mediate the early recognition of pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs). TLR signaling modulates the tumor microenvironment from “cold” to “hot” leading to immune sensitization of tumor cells to treatments and improved patient prognosis. In addition, TLR signaling activates the adaptive immune response to improve the response to cancer immunotherapy through the regulation of anti-tumor T cell activity. This review will highlight recent progress in our understanding of the role of TLRs in lung cancer immunity and immunotherapy. Toll-like receptors (TLRs) recognize both pathogen-associated molecular patterns (PAMPs) and endogenous damage-associated molecular patterns (DAMPs). Ample evidence has demonstrated that TLRs play a critical role in cancer development and treatment. TLR signaling activation not only triggers innate immune responses but also regulates adaptive immunity responses. As such, TLRs and TLR signaling serve as targets for monitoring cancer progression and the development of a new strategy for cancer treatment. Lung cancer has been the leading cause of cancer-related deaths since the 1950s in men and the 1980s in women, surpassing breast cancer . HoweverThe host immune response relies on the actions of both innate and adaptive immunity, but the importance of innate immunity was not fully recognized until the discovery of TLRs approximately 25 years ago . These rTLRs are type I integral transmembrane proteins, including the leucine-rich repeat (LRR) domain, a transmembrane domain, and the Toll-interleukin receptor (TIR) domain . The LRRLung cancer is currently the world’s leading cause of cancer-related deaths . These dVast amounts of evidence have demonstrated that TLRs play an important role in lung cancer development and treatment. However, opposing roles for either pro- or anti-tumor progression have been described for different TLRs Figure 1Lung DCs can be divided into either conventional or plasmacytoid DCs (pDCs) . DCs havUnder normal conditions, macrophages are seen in high numbers within the lung TME. TLR2, TLR3, TLR4, and TLR6 are expressed at higher levels than TLR7 and TLR9 in lung macrophages . Upon liThe activation of TLR2 signaling in MDSCs induces tumor regression. TLR2 and TLR 7/8 are highly expressed in monocytic-MDSCs (m-MDSCs) making them an important aid for tumor immune evasion  cells are important innate immune cell effectors used against tumor cells. They are activated by DCs, and their cytotoxicity is induced through the MyD88-independent pathway. Mast cells are another innate immune cell that abundantly surrounds solid tumors and are known to be pro-tumor by enhancing tumor angiogenesis . Innate  evasion , 44. Fur evasion .Cytotoxic T lymphocytes are the major adaptive cellular effectors against tumor regression. Tumor regression is dependent on the MyD88-dependent signaling pathway involved in the expression of MHC-I. Humoral immunity encompasses the adaptive immune response that occurs through B lymphocyte antibody production. Normally the B cell numbers in the lung are considerably low, but significantly increased in lung cancer . TLR7 anin vivo murine model showed a significant reduction in macrophage infiltration and inflammatory cytokine expression in TLR2 knockout mice compared to wild type (WT), revealing the necessity of TLR2 recognition of a cancer cell ligand for promotion of metastatic growth  for liver cancer and HPV (Human Papillomavirus) for cervical cancer. It is not surprising that TLRs recognize these viruses and play a crucial role in pathogen-associated oncogenesis. A recent study reported that bacteria-associated cancers are much higher than 20%, and that bacterial footprints like LPS can be detected in about 60-70% of some cancers . Thus, tg cancer . Additiog cancer . Many reg cancer –96, but g cancer , 98. HowTLRs recognize not only pathogens but also endogenous danger molecules, particularly those derived from cancer cells. Recent structural analyses have identified novel binding amongst TLRs-PAMPs , 100 HowWe posit that TLRs are ideal targets for therapeutic development. Some TLRs can induce anti-tumor activity, while others can regulate adaptive immunity. The resistance of cancer patients to immunotherapy has been associated with an immunologically “cold” TME where immune-killing activity is repressed . The actTLRs are now being targeted for drug development and immunotherapy discovery because of promising innate immune activation. The usefulness of this approach is limited at this time due to conflicting pro- and anti-tumor activities of TLR receptors. The opposing effects of TLRs appear to be cell type- and microenvironment-specific, but the molecular determinants of these biological responses are not yet understood. Encouraging results from clinical studies using TLRs alone or in combination with other checkpoint inhibitors are beginning to appear. Therefore, there is a need to expand this field of research by exploring TLRs and their corresponding agonists and antagonists as promising tools in immunotherapy discovery and drug development.DZ and BH designed and outlined this review article; BH performed literature search and wrote the draft of the manuscript. All authors edited and approved the final manuscript.This study was partially supported by Texas A&M University, a grant from the Cancer Prevention & Research Institute of Texas , and a NIH grant (R21CA176698).We appreciate the help from all our contributing authors in writing this review. We also want to acknowledge Texas A&M University for providing access to journals and the use of BioRender.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Refugees are forced migrants but there is a large variation in the distance that refugees cover and there is a knowledge gap on how this may affect refugees’ health and health care needs.Herein, we investigate the association between long-distance migration and post-traumatic stress disorder (PTSD), a serious psychiatric disorder associated with deteriorating mental and somatic health and highly prevalent in refugees.Included were 712 adult Syrian refugees and asylum seekers in Lebanon and Denmark arriving no more than 12 months prior to inclusion. The Harvard Trauma Questionnaire was used to assess PTSD and the estimate of association was obtained by multiply imputing missing data and adjusting for confounding by propensity score-weighting with covariates age, sex, socioeconomic status, trauma experience, and WHO-5-score, reporting the bootstrap 95-percentile confidence interval (95% CI). Additionally, a number of sensitivity analysis were carried out.The prevalence of PTSD was high in both Lebanon (55%) and Denmark (60%) and long-distance migration was associated with a 9 percentage point  increase in the prevalence of PTSD among newly arrived Syrian refugees and asylum-seekers.In the present study the prevalence of PTSD increased after long-distance migration which may support considering “long-distance migration” in refugee health screenings and in particular when assessing the risk of post-traumatic stress disorder. This is a first step in examining the health effects of migration on refugee health.No significant relationships.


Conduction disturbances are an important complication of ST-elevation myocardial infarction (STEMI). Conduction disturbances such as fascicular blocks and bundle branch blocks are associated with alteration of QRS morphology and secondary ST-T wave changes that can influence the diagnosis of acute myocardial ischemia. We report an interesting case where a patient presented with inferior wall myocardial infarction (MI), right bundle branch block (RBBB), and left anterior hemiblock (LAHB). We discuss the challenges in diagnosing MI in such patients, including the impact of QRS changes in RBBB and LAHB, their influence on diagnosis of STEMI, and differentiation of combined first-degree AV block and bifascicular block from trifascicular block. Conduction abnormalities such as fascicular blocks and bundle branch blocks lead to changes in the QRS morphology and produce secondary ST-T wave changes. Conduction abnormalities are well-known complications of acute myocardial infarction (MI). However, conduction disturbance may be pre-existent in patients with suspected MI and can mask or simulate the electrocardiographic diagnosis of MI. In the present case, a patient with inferior wall MI also had a right bundle branch block (RBBB) and left anterior hemiblock (LAHB) in the electrocardiogram (ECG). We address the various challenges in the diagnosis of MI in patients with fascicular block and bundle branch blocks.A 70-year-old man presented with precordial chest discomfort for three hours. There was no history of any cardiovascular illness in the past. The patient did not have cardiovascular risk factors such as hypertension, diabetes, smoking, or dyslipidemia. The vitals were stable, and the cardiovascular system examination was unremarkable. The ECG at presentation is shown in Figure The ECG shows RBBB with left axis deviation. In the absence of a previous ECG, it is difficult to ascertain if the RBBB is new-onset or pre-existing. The presence of qR complexes in I, aVL, and rS complexes in the inferior leads is consistent with LAHB. The ST-segment elevation measuring > 1 mm is present in leads II, III, and aVF with reciprocal changes in other leads. These features are diagnostic of inferior wall MI. The ST-segment depression in leads V1-V2 with an R/S ratio >1 indicates posterior wall MI. The PR interval measures 200 milliseconds. Considering the diagnosis of acute ST-segment elevation myocardial infarction (STEMI) and an anticipated delay in transfer to a hospital with a primary percutaneous coronary intervention (PCI) facility, we administered thrombolytic therapy with intravenous tenecteplase. The chest discomfort was resolved and ST-T changes in the ECG were indicative of successful thrombolysis  with multiple posterolateral ventricular branches supplying the inferior and inferolateral left ventricular wall. The culprit lesion was noted in the proximal vessel Figure . SuccessConduction disturbances such as bundle branch blocks cause secondary ST-T changes which can influence the diagnosis of acute myocardial ischemia . MyocardThe QRS changes in LAHB can mask or simulate the QRS changes of MI ,4. The LThe LAHB may simulate old anterior wall MI by producing q waves or QS complexes in the precordial leads. The LAHB directs the initial depolarization vector posteriorly, resulting in q waves in leads V2 and V3 ,7. Such Uncomplicated RBBB can result in ST-T changes in lead V1. These include ST-segment depression and the presence of inverted T waves . HoweverIt is difficult to ascertain whether the bifascicular block in the present case occurred as a complication of MI, or was present at baseline. Both the right bundle and left anterior fascicle have a common blood supply from the septal branches of the left anterior descending artery; hence, RBBB and LAHB often co-exist . While tThe PR prolongation in the setting of RBBB and LAHB is not diagnostic of a trifascicular block as nearly half of these patients have a delay in the AV node . The surConduction disturbances can cause significant ST-T changes so as to influence the diagnosis of acute MI. The QRS changes in LAHB can mask or simulate the QRS changes of previous MI. The LAHB and uncomplicated RBBB alone do not cause widespread secondary ST-segment changes and generally do not confound the diagnosis of STEMI. The PR prolongation with concomitant RBBB and LAHB is not diagnostic of a trifascicular block as it is difficult to differentiate from the surface ECG if the conduction delay is nodal or infranodal.


Correction to: Affective Science10.1007/s42761-022-00112-xAsymmetrical Effects of Sleep and Emotions in Daily Life.” Specifically, they did not copy some of the equations correctly and failed to convert some of the Greek symbols.The publishing team at Affective Science made errors in the manuscript “This erratum is presented to fix these errors.The original article has been corrected.


However, integrating information from different experiments remains challenging because of the batch effect generated by various technological and biological variations in the transcriptome. Numerous batch-correction methods to deal with this batch effect have been developed in the past. However, a user-friendly workflow to select the most appropriate batch-correction method for the given set of experiments is still missing. We present the SelectBCM tool that prioritizes the most appropriate batch-correction method for a given set of bulk transcriptomic experiments, improving biological clustering and gene differential expression analysis. We demonstrate the applicability of the SelectBCM tool on analyses of real data for two common diseases, rheumatoid arthritis and osteoarthritis, and one example to characterize a biological state, where we performed a meta-analysis of the macrophage activation state. The R package is available at  Over the last two decades, high-throughput transcriptome studies have resulted in a vast accumulation of bulk gene-expression experiments in public repositories and value-added resources, including microarray and RNA-seq data . The potHowever, there are several challenges before fully harnessing this wealth of data for scientific investigation; one of the most critical is the ‘batch effect’ in collated transcriptome datasets ,11. BatcGiven the tremendous potential buried in the available bulk transcriptome datasets, we aimed to develop a freely available R-based tool to facilitate gene-expression integration. Here, we present the SelectBCM tool to choose the appropriate BCM for the given set of bulk transcriptomic data. Integrating different BCM tools and assessment strategies into the same framework, the SelectBCM tool, will increase the reusability and reproducibility of these BCMs for batch correction and allow the user to compare and select the top BCM for their chosen datasets. We also demonstrate the application of SelectBCM through case studies and compare differentially expressed genes (DEGs) obtained after batch correction with the traditional meta-analysis approach.Homo sapiens  tool to estimate the batch effect and biological variability of interest in the SelectBCM tool in a given meta-experiment . In summs(i) can range from –1 to 1, with s as an indicator of the presence or absence of ‘batch effects’.To assess the clustering pattern of biosamples before and after batch correction, we compute the silhouette coefficient . TheoretTo estimate the linear batch effect, we perform linear regression using the top 20 principal components (PCs) obtained from PCA as a default setting against a batch  variable. For the estimation purpose, we use the scaled pcRegression score by scaling it by the total variance of the top 20 PCs, as suggested in the κBET package .We compute the regional entropy of batch mixing before and after batch correction using the first two PCs, as indicated in the MNN package . In our Highly variable genes (HVGs) contribute to cell-to-cell variation within a homogeneous cell population and represent true biological heterogeneity ,24. We qTo rank BCMs, we compute sumRank using results obtained from evaluation steps as parameters Figure . In the PVCA: batch  for visualization purposes. We then compute sumRank using the evaluation matrix explained below:where Therefore, sumRank provides the final ranking of BCMs for the given meta-experiment in descending order. BCM ranked higher across evaluation methods will get a higher weightage and, therefore, first rank among others (Output 3).We used visual and DEG analysis approaches to qualitatively assess the impact of the top 3 BCMs on biological inferences. To visually assess the impact of batch correction on biological clustering, we applied PCA on a meta-experiment obtained after batch correction and compared how well biosamples from different batches were mixed. We perform DEG analysis to assess the impact of the batch correction on identifying relevant biological signals. We also compare DEGs identified by BCMs with those determined by the traditional meta-analysis approach (MetaVolcanoR), where people study each experiment individually and then statistically combine the list of significant genes . We provhttps://github.com/ebi-gene-expression-group/selectBCM) implements a flexible framework in R to select the best BCMs for transcriptomic experiments  and osteoarthritis (OA) diseases as examples to demonstrate the usability of SelectBCM for microarray experiments. We curated microarray experiments for RA and OA from synovial fluid or bone specimens for For RA disease, we selected E-GEOD-1919, E-GEOD-55235, E-GEOD-48780, E-GEOD-12021, E-GEOD-55457 and E-MTAB-3201 experiments contributing at least RA or control synovial specimen samples or both. SelectBCM performs batch correction and evaluation for the RA meta-experiment. We observe that the evaluation methods did not produce a consensus among themselves to identify the best BCM Figure . Even thTo assess the impact of the top 3 BCMs shortlisted by the SelectBCM on biological clustering, we carried out a PCA Figure . We firsWe assess the impact of the batch correction on differential gene analysis by comparing DEGs obtained after applying the top 3 BCMs with DEGs obtained from the traditional meta-analysis approach. As expected, different BCMs resulted in different sets of DEGs having minimal overlap  Figure . We alsocis-regulatory region sequence-specific DNA binding, RNA polymerase II transcription regulatory region sequence-specific DNA binding and double-stranded DNA binding were the top 5 downregulated molecular functions among top BCMs and MetaVolcanoR  and downregulated (13 genes) using the EnrichR tool . The topR Figure  30). Ev. Evcis-rFinally, we also checked the tissue specificity of these common DEGs using the Jensen tissue score from EnrichR . Jensen Based on biological clustering, DEG recovery, molecular functions and tissue specificity of common DEGs, we concluded that the top 3 BCMs could capture biologically relevant processes while retaining tissue-specific information for the RA study. However, depending on the choice of BCM, some methods (such as limma) will require a more stringent statistical cut-off to filter DEGs.We performed a similar analysis for OA and provided a detailed study description in in vitro monocyte-derived macrophage (MDM) activation state models as a case study to demonstrate the application of the SelectBCM tool for bulk RNA-seq experiments. We choose the MDM study as macrophages represent heterogeneous immune cell populations in the context of tissue and diseases, and most studies use in vitro MDMs to understand the stimulant-associated heterogeneity , we again observed that all three BCMs captured the right direction of differential expression changes of marker genes of the IL-4 stimulation but failed to capture it in the uncorrected meta-experiment Figure . HoweverBelow we discuss the key challenges and limitations of performing batch correction in the bulk transcriptome. This study has emphasized the challenges associated with how biological heterogeneity interferes with the batch-correction outcome. Through a meta-analysis study of systemic juvenile idiopathic arthritis , we demonstrate a scenario where biological heterogeneity (active and inactive states of illness) is present, the study design is unbalanced and the sample size is small. We observe that clustering did not improve after batch correction in both studies . We alsoWe designed the SelectBCM tool to solve the old problem of selecting the suitable BCM for bulk transcriptomic experiments. We implemented >9 state-of-the-art batch-correction algorithms in our workflow designed to handle bulk transcriptomes from microarray and RNA-seq technology. We showcase the useability of our workflow to address the problem of choosing the most appropriate BCM and the importance of having multiple evaluation methods. Using RA and OA datasets, we observed that Q_ComBat and ComBat1 were the top BCMs for better DEG recovery, and downstream biological inferences agreed with the traditional meta-analysis approach. For the bulk RNA-seq experiment, we observed that ComBat_seq is better for retaining biological signals. We expect our tool will speed up the bulk transcriptomic data integration process, starting from filtering the datasets based on metadata to batch correction and allowing users to focus on deciphering biological signals.https://hub.docker.com/r/yysong123/selectbcm. All datasets and analyses used in the paper are public, referenced and downloadable at https://doi.org/10.5281/zenodo.7482141.The docker image of the tool is available at lqad014_Supplemental_FileClick here for additional data file.


Personal protection equipment (PPE) use in hospitals has consistently increased due to the Sars-Cov-2 outbreak, in wards repurposed for Covid-19 patients and wards that kept their usual activity. This increase influenced an environmental emergency in terms of health waste (HW) disposal. This study aims to assess the economic and environmental impact of the increase in HW generated before and during the pandemic in an Italian Hospital.Data from 2016 to 2019 and 2020 to 2021 was retrieved from Risk Management department. Per capita and per days-of-stay waste quantity were calculated for the hospital inpatient wards and medical service areas . Linear regression models assessed the epidemiological impact of COVID, and LOESS analysis modeled the relationship between infectious HW generation and the percentage of COVID-related inpatient days. Average weight of HW per patient was used to estimate the monetary value of CO2 produced.Preliminary results show that the inpatient days related to COVID nonlinearly influenced the infectious HW generated by wards. PPE usage increased in every context, and the proportion of COVID-related bed-days ranged from 2% to 12% in low-incidence months to 17% to 31% during acute phases. Pre-COVID CO2 production weighted 487 kg per patient and cost 1705€ per-capita, whereas during the pandemic it amounted to 768 kg per patient and cost 2688€ per capita which resulted in a significant increase of 983€ per patient.In light of the results, HW disposal is an urgent issue that should be addressed by policy makers when implementing new monitoring systems for hospitals. A more adequate disposal of HW could substantially contribute in reducing air pollution and concurrently reduce the economic impact health systems due to the coronavirus pandemic.The COVID-19 pandemic has brought an increase in hospital waste generation.The improvement of HW disposal monitoring system should be considered to avoid reaching critical levels in the near future.


Mycobacterium tuberculosis (Mtb) topoisomerase I (TopA) is the only type IA topoisomerase in this organism and has been validated as an essential target for TB drug discovery. Toxin-antitoxin (TA) systems participate as gene regulators within bacteria. The TA systems contribute to the long-term dormancy of Mtb within the host-cell environment. Mtb’s toxin MazF4 (Rv1495) that is part of the MazEF4 TA system has been shown to have dual activities as endoribonuclease and topoisomerase I inhibitor. We have developed a complementary assay using an Escherichia coli strain with temperature-sensitive topA mutation to provide new insights into the MazF4 action. The assay showed that E. coli is not sensitive to the endoribonuclease activity of Mtb MazF4 but became vulnerable to MazF4 growth inhibition when recombinant Mtb TopA relaxation activity is required for growth. Results from the complementation by Mtb TopA mutants with C-terminal deletions showed that the lysine-rich C-terminal tail is required for interaction with MazF4. Site-directed mutagenesis is utilized to identify two lysine residues within a conserved motif in this C-terminal tail that are critical for MazF4 inhibition. We performed molecular dynamics simulations to predict the Mtb TopA-MazF4 complex. Our simulation results show that the complex is stabilized by hydrogen bonds and electrostatic interactions established by residues in the TopA C-terminal tail including the two conserved lysines. The mechanism of Mtb TopA inhibition by MazF4 could be useful for the discovery of novel inhibitors against a new antibacterial target in pathogenic mycobacteria for treatment of both TB and diseases caused by the non-tuberculosis mycobacteria (NTM).Only about half the multi-drug resistant tuberculosis (MDR-TB) cases are successfully cured. Thus, there is an urgent need of new TB treatment against a novel target.  Mycobacterium tuberculosis(Mtb) is the microorganism that causes Tuberculosis (TB), a leading infectious disease in the world with 10.0 million individuals experiencing active-TB, while 1.5 million succumbed in 2020  as the only type I topoisomerase along with gyrase as the only type II topoisomerase. Gyrase is well known as an important target for fluoroquinolones  that form the active site for cleaving and rejoining of a single-strand of DNA  that can bind ssDNA  of toxin-antitoxin (TA) systems identified in the host . The toxthe host , 2008. Aactivity  through d of DNA . The mord of DNA  also connd ssDNA , and a lactivity . The inhMtb TopA coding sequence in strain H37Rv into pLIC-HK vector .Cloning of K vector  to generK vector . The cloeviously . The Mazeviously  in the pE. coli XL1-Red strain with mutS, mutD, and mutT mutations (from Agilent). The pLIC-MTOP plasmid extracted from XL1-Red was amplified in E. coli XL1-Blue. The coding region for TopA was excised with restriction enzymes BamHI and NcoI and ligated to the vector backbone generated from digestion of the original plasmid. The mutant pLIC-MTOP plasmid library was obtained from transformation of XL1-Blue with the ligation product.A library of mutant pLIC-MTOP plasmid with random mutations  in the TMtb TopA terminating after residue 840 or 910 were made by assembly cloning of vector and insert fragments generated with primers listed in Mtb TopA were later introduced into pLIC-MTOP plasmid using the NEB HiFi mutagenesis Master Mix and primers listed in pLIC-MTOP plasmids that express truncated version of Escherichia coli strain AS17 topA17(am) and rsupD43,74Tet with temperature sensitive topA is not viable at 42°C  as primary antibody (™ reversible stain) to confirm for equal loading and gel transfer of proteins.Western blot analysis using rabbit polyclonal antibodies raised against antibody  and goatMtb TopA (MTOP-FL) structure using RaptorX  or Mtb TopA (from plasmid pLIC-MTOP). When an additional plasmid pBAD-Rv1495 expressing the MazF4 toxin was present in the cell, growth complementation by recombinant E. coli TopA was not affected, while growth of AS17 complementation by recombinant Mtb TopA was reduced by up to 103-fold. The CFU/ml values were measured in three replicated experiments to confirm the reduction of growth complementation by Mtb TopA by > 200-fold due to the presence of the Mtb MazF4 toxin  is composed of two main domains: N-terminal domain and C-terminal domain  and complete MazF4 dimer. We performed MD simulations to access stability and structural integrity of the docked complex  by Mtb MazF4  and LIC-MTOP-910 t  reduced the degree of growth complementation at 42°C because the C-terminal subdomains and C-terminal tail play important roles in interaction with DNA substrate during relaxation of supercoiled DNA , Colciencias (Convocatoria 586 de becas en el exterior to PG). PBT acknowledges the computational resources acquired with a research restart discretionary fund from the office of the Dean for Research at Georgetown University Medical Center.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.https://www.frontiersin.org/articles/10.3389/fmicb.2022.1032320/full#supplementary-materialThe Supplementary material for this article can be found online at: Click here for additional data file.