SARS-CoV-2 inactivation by supercritical carbon dioxide coupled with hydrogen peroxide

The recent COVID-19 pandemic has underscored the need for innovative decontamination techniques capable of treating sensitive materials potentially contaminated. Combining supercritical carbon dioxide (scCO2) with sterilant agents has shown promise in this regard. This study aimed at testing scCO2 as a virus inactivation method for biomedical materials contaminated with the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2). The virus was inoculated on a stainless-steel carrier and treated at 45 °C and 8 MPa. No inactivation was detected when only scCO2 was used, even after long treatment times (60 min). The addition of 50 ppm of H2O2 to the process allowed the inactivation of more than 5 Log PFU (Plaque Forming Unit) of the virus by only pressurising and depressurising the vessel, while a 20-min process is needed by only using H2O2. Overall, the study demonstrates a synergistic effect when H2O2 is added to the scCO2 process for the inactivation of SARS-CoV-2

An Integrated Meta-Analysis of Two Variants in HOXA1/HOXB1 and Their Effect on the Risk of Autism Spectrum Disorders

BACKGROUND: HOXA1 and HOXB1 have been strongly posed as candidate genes for autism spectrum disorders (ASD) given their important role in the development of hindbrain. The A218G (rs10951154) in HOXA1 and the insertion variant in HOXB1 (nINS/INS, rs72338773) were of special interest for ASD but with inconclusive results. Thus, we conducted a meta-analysis integrating case-control and transmission/disequilibrium test (TDT) studies to clearly discern the effect of these two variants in ASD. METHODS AND FINDINGS: Multiple electronic databases were searched to identify studies assessing the A218G and/or nINS/INS variant in ASD. Data from case-control and TDT studies were analyzed in an allelic model using the Catmap software. A total of 10 and 7 reports were found to be eligible for meta-analyses of A218G and nINS/INS variant, respectively. In overall meta-analysis, the pooled OR for the 218G allele and the INS allele was 0.97 (95% CI = 0.76-1.25, P(heterogeneity) = 0.029) and 1.14 (95% CI = 0.97-1.33, P(heterogeneity) = 0.269), respectively. No significant association was also identified between these two variants and ASD risk in stratified analysis. Further, cumulative meta-analysis in chronologic order showed the inclination toward null-significant association for both variants with continual adding studies. Additionally, although the between-study heterogeneity regarding the A218G is not explained by study design, ethnicity, and sample size, the sensitive analysis indicated the stability of the result. CONCLUSIONS: This meta-analysis suggests the HOXA1 A218G and HOXB1 nINS/INS variants may not contribute significantly to ASD risk

The in vitro antiviral activity of Tumor Necrosis Factor (TNF) in WISH cells is mediated by IFN-beta induction

We present evidence showing that TNF is capable of inducing an antiviral state in WISH cells thereby protecting them from the cytopathic effect of vesicular stomatitis virus. Establishment of the antiviral state requires pretreatment with TNF. Such pretreatment not only protects the cells in a dose-dependent manner, but it markedly reduces virus yield as well. Kinetic studies have shown that a pretreatment period as short as 4 h at 37 degrees C is effective in conferring protection. The antiviral activity of TNF could be attributed to the induction of IFN-beta. In fact, polyclonal antibodies to IFN-beta completely neutralized the antiviral state elicited by TNF. 2-5A synthetase activity was significantly enhanced when the cells were treated with doses of TNF that afforded antiviral protection. Finally, addition of specific antibodies to IFN-beta 2 (IL-6) during TNF pretreatment failed to abolish the antiviral state, thus suggesting that IFN-beta 2 is not involved in the TNF-induced antiviral state. Also, a homogeneous IFN-beta 2 preparation failed to exert antiviral activity in our cell system

Lack of growth inhibitory synergism with combined MAPK/PI3K inhibition in preclinical models of pancreatic cancer

Despite substantial advances in chemotherapy and biology understanding, pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest solid tumors. Attempts at exploiting PDAC biology for therapeutic purposes have failed and the likelihood of approval for new agents that enter phase I testing in this disease is down to a dismal 2.3%[1]. Recently, a combination of MEK (selumetinib) and AKT (MK-2206) inhibitors failed to demonstrate clinical benefit in unselected PDAC patients[2], adding to a long list of targeted agents that have failed clinical testing (EGFR/VEGFR, SMO, and Notch inhibitors, to name a few). We thus asked ourselves whether such failure could have been predicted preclinically. We explored pharmacologic interactions between MEK inhibitors (trametinib) and PI3K pathway inhibitors [gedatolisib (PI3K/mTOR inhibitor) andMK-2206 (AKT inhibitor)] in vitro, using 6 human PDAC cell lines and the “normal pancreatic epithelium” cell line HPDE. Single-agent inhibition of MEK, PI3K/mTOR, or AKT inhibited cell growth to a variable extent in all cell lines examined. However, combined inhibition of MEK and PI3K/mTOR (trametinib/gedatolisib) afforded frankly antagonistic effects in Panc1, MiaPaCa2, T3M4, PaCa44, and HPDE, slightly additive effects in HPAFII, and synergistic effects only in L3.6pl cells (Fig. 1A); similarly, combined MEK/AKT inhibition (trametinib/MK-2206) was antagonistic in all cell lines tested (Fig. 1A-C). Overall, no growth inhibitory synergism in vitro was observed in any of the cell lines tested, with the exception of L3.6pl cells in response to trametinib/gedatolisib combination. Our group has recently shown that combined inhibition of the MAPK and PI3K pathways affords synergistic anti-tumor effects almost exclusively in cancer cells without a functional PTEN gene/protein (PTEN-loss)[3]. We thus examined PTEN expression in the panel of PDAC cell lines examined: no PTEN mutations or bi-allelic loss have been reported for these cells and all displayed detectable levels of PTEN protein (Fig 1A, D), thus falling in the PTEN-competent category according to the definition recently proposed by our group[3]. Consistent results (lack of growth inhibitory synergism) had, indeed, already been obtained in the PTEN-competent cell lines HPAFII and MiaPaCa2, using another combination of MEK and mTOR inhibitors (trametinib and everolimus)[3]. Inactivating PTEN point mutations or LOH rarely occur in human PDAC[4]; thus, based on the preclinical data presented here, the failure of selumetinib/MK-2206 to achieve clinical benefit in unselected PDAC patients would have been largely anticipated. Extensive preclinical modeling and early selection biomarker development are, in our opinion, crucial to successful drug development, in addition to uniform trial eligibility criteria, stringent statistical methods, and detection of robust activity signals in early phase trials. Unfortunately, such rules have been often overlooked in advanced PDAC, resulting in the identification of only 3 clinically meaningful agents or combinations out of 32 phase III trials enrolling >13,000 patients[5]. This line of reasoning especially applies to combined pathway inhibition, which implies increased monetary and toxicity costs, representing a high risk for all stakeholders, should it fail to demonstrate more than additive benefits, and to advanced PDAC, a disease setting in which novel effective therapeutic approaches are urgently needed

Lack of association between serotonin transporter gene promoter variants and autistic disorder in two ethnically distinct samples

Family-based studies performed to date provide conflicting evidence of linkage/association between autistic disorder and either the 'short' [Cook et al., 1997: Mol Psychiatry 2:247-250] or the 'long' [Klauck et al., 1997: Hum Mol Genet 6:2233-2238] allele of a polymorphic repeat located in the serotonin transporter (5-HTT) gene promoter region, affecting 5-HTT gene expression [Lesch et al., 1996: Science 274:15271531]. The present study was designed to assess linkage and linkage disequilibrium in two new ethnically distinct samples of families with primary autistic probands. The 5-HTT promoter repeat was genotyped in 54 singleton families collected in Italy and in 32 singleton and 5 multiplex families collected in the U.S.A., yielding a total sample of 98 trios. Linkage/association between 5-HTT gene promoter alleles and autistic disorder was assessed using the transmission/disequilibrium test (TDT) and the haplotype-based haplotype relative risk (HHRR). Both the Italian and the American samples, either singly or combined, displayed no evidence of linkage/association between 5- HTT gene promoter alleles and autistic disorder. Our findings do not support prominent contributions of 5-HTT gene variants to the pathogenesis of idiopathic infantile autism. Heterogeneity in pathogenetic mechanisms underlying the disease may require that linkage/association studies be targeted toward patient subgroups isolated on the basis of specific biochemical markers, such as serotonin (5-HT) blood levels. (C) 2000 Wiley- Liss, Inc

Lack of association between serotonin transporter gene promoter variants and autistic disorder in two ethnically distinct samples

Family-based studies performed to date provide conflicting evidence of linkage/association between autistic disorder and either the 'short' [Cook et al., 1997: Mol Psychiatry 2:247-250] or the 'long' [Klauck et al., 1997: Hum Mol Genet 6:2233-2238] allele of a polymorphic repeat located in the serotonin transporter (5-HTT) gene promoter region, affecting 5-HTT gene expression [Lesch et al., 1996: Science 274:15271531]. The present study was designed to assess linkage and linkage disequilibrium in two new ethnically distinct samples of families with primary autistic probands. The 5-HTT promoter repeat was genotyped in 54 singleton families collected in Italy and in 32 singleton and 5 multiplex families collected in the U.S.A., yielding a total sample of 98 trios. Linkage/association between 5-HTT gene promoter alleles and autistic disorder was assessed using the transmission/disequilibrium test (TDT) and the haplotype-based haplotype relative risk (HHRR). Both the Italian and the American samples, either singly or combined, displayed no evidence of linkage/association between 5- HTT gene promoter alleles and autistic disorder. Our findings do not support prominent contributions of 5-HTT gene variants to the pathogenesis of idiopathic infantile autism. Heterogeneity in pathogenetic mechanisms underlying the disease may require that linkage/association studies be targeted toward patient subgroups isolated on the basis of specific biochemical markers, such as serotonin (5-HT) blood levels. (C) 2000 Wiley- Liss, Inc