BACKGROUND

Bronchiolitis caused by the respiratory syncytial virus (RSV) in infants less than two years old is a growing public health concern worldwide, and there is currently no safe and effective vaccine. A major component of RSV nucleocapsid, the nucleoprotein (N), has been so far poorly explored as a potential vaccine antigen, even though it is a target of protective anti-viral T cell responses and is remarkably conserved between human RSV A and B serotypes. We recently reported a method to produce recombinant N assembling in homogenous rings composed of 10-11 N subunits enclosing a bacterial RNA. These nanoparticles were named sub-nucleocapsid ring structure (N SRS).

RESULTS

The vaccine potential of N SRS was evaluated in a well-characterized and widely acknowledged mouse model of RSV infection. BALB/c adult mice were immunized intranasally with N SRS adjuvanted with the detoxified E. coli enterotoxin LT(R192G). Upon RSV challenge, vaccinated mice were largely protected against virus replication in the lungs, with a mild inflammatory lymphocytic and neutrophilic reaction in their airways. Mucosal immunization with N SRS elicited strong local and systemic immunity characterized by high titers of IgG1, IgG2a and IgA anti-N antibodies, antigen-specific CD8(+) T cells and IFN-gamma-producing CD4(+) T cells.

CONCLUSIONS

This is the first report of using nanoparticles formed by the recombinant nucleocapsid protein as an efficient and safe intra-nasal vaccine against RSV.

(<em>b</em>)Background:</<em>b</em>) In patients with type 2 dia<em>b</em>etes mellitus (T2DM), sodium-glucose cotransporter 2 (SGLT2) inhi<em>b</em>itors reduce the risk of hospitalization for heart failure (HHF). We assessed the effect of ertugliflozin on HHF and related outcomes. (<em>b</em>)Methods:</<em>b</em>) VERTIS CV, a dou<em>b</em>le-<em>b</em>lind, place<em>b</em>o-controlled trial, randomized patients with T2DM and atherosclerotic cardiovascular (CV) disease to once-daily ertugliflozin 5 mg, 15 mg or place<em>b</em>o. Prespecified secondary analyses compared ertugliflozin (pooled doses) versus place<em>b</em>o on time to first event of HHF and composite of HHF/CV death, overall and stratified <em>b</em>y prespecified characteristics. Cox proportional hazard modeling was used with the Fine and Gray method to account for competing mortality risk, and Andersen-Gill modeling to analyze total (first+recurrent) HHF and total HHF/CV death events. (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) 8246 patients were randomized to ertugliflozin (n=5499) or place<em>b</em>o (n=2747); n=1958 (23.7%) had a history of heart failure (HF) and n=5006 (60.7%) had pre-trial ejection fraction (EF) availa<em>b</em>le, including n=959 with EF≤45%. Ertugliflozin did not significantly reduce first HHF/CV death (HR, 0.88; 95% CI, 0.75, 1.03). Overall, ertugliflozin reduced risk for first HHF (HR, 0.70; 95% CI, 0.54, 0.90; <i>P</i>=0.006). Prior HF did not modify this effect (HF: HR, 0.63; 95% CI, 0.44, 0.90; no HF: HR, 0.79; 95% CI, 0.54, 1.15; <i>P</i> interaction=0.40). In patients with HF, the risk reduction for first HHF was similar for those with reduced EF≤45% vs preserved EF>45% or unknown. However, in the overall population, the risk reduction tended to <em>b</em>e greater for those with EF≤45% (HR, 0.48; 95% CI, 0.30, 0.76) versus EF>45% (HR, 0.86; 95% CI, 0.58, 1.29). Effect on risk for first HHF was consistent across most su<em>b</em>groups, <em>b</em>ut greater <em>b</em>enefit of ertugliflozin was o<em>b</em>served in three populations with <em>b</em>aseline eGFR&<em>lt</em>;60mL/min/1.73m², al<em>b</em>uminuria, and diuretic use (each <i>P</i> interaction&<em>lt</em>;0.05). Ertugliflozin reduced total events of HHF (RR, 0.70; 95% CI, 0.56, 0.87) and total HHF/CV death (RR, 0.83; 95% CI, 0.72, 0.96). (<em>b</em>)Conclusions:</<em>b</em>) In patients with T2DM with or without <em>b</em>aseline HF, ertugliflozin reduced risk for first and total HHF and total HHF/CV death, adding further support for the use of SGLT2 inhi<em>b</em>itors in primary and secondary prevention of HHF. (<em>b</em>)Clinical Trial Registration:</<em>b</em>) URL: https://clinica<em>lt</em>rials.gov Unique Identifier: <a href="http://clinica<em>lt</em>rials.gov/show/NCT01986881" title="See in ClinicalTrials.gov">NCT01986881</a>.

Objective: The aim of this manuscript is to investigate transversally Ear Nose Throat (ENT) symptoms COVID-19 infection correlated and to study the neurotropism and neuroinvasiveness of the virus in the head-neck district through the investigation of the sense of smell, taste, tearing, salivation and hearing.

Methods: A total of 50 patients with laboratory-confirmed COVID-19 infection were included in our study. For each patient we evaluated the short version of the Questionnaire of Olfactory Disorders-Negative Statements (sQOD-NS), the Summated Xerostomia Inventory-Dutch Version (SXI-DV), The Standardized Patient Evaluation of Eye Dryness (SPEED), Schirmer test I, the Hearing Handicap Inventory For Adults (HHIA) and the Tinnitus Handicap Inventory (THI). All the tests we carried out were performed during the active phase of the symptomatology from COVID-19 (Condition A) and 15 after SARS-COV-2 RT-PCR test negative (Condition B).

Results: A total of 46 patients (92%) had olfactory dysfunction related to the infection. The 70% of patients reported gustatory disorders. Cough, fever, headache and asthenia were the most prevalent symptoms. There was a statistically significant difference (p < 0,001) in sQOD-NS, SXI-DV, SPEED, Schirmer test, HHIA and THI between Condition A and Condition B.

Conclusions: In our population there was an alteration of the sense of taste, of the sense of smell, dry eyes and of the oral cavity and an auditory discomfort, symptoms probably linked to the neurotropism of the virus. Furthermore, anosmia, dysgeusia and xerostomia are early symptoms of COVID-19, which can be exploited for an early quarantine and a limitation of viral contagion.

Keywords: Anosmia; COVID-19; Dry eye; ENT; Head and neck; Hearing loss; Neurotropism; SARS-CoV-2.

<p><div>(<em>b</em>)BACKGROUND</<em>b</em>)</div>A large proportion of patients with a SARS-Cov-2-associated respiratory failure develop an acute respiratory distress syndrome (ARDS). It has <em>b</em>een recently suggested that SARS-Cov-2-associated ARDS may differ from usual non-SARS-Cov-2-associated ARDS <em>b</em>y higher respiratory system compliance (C<su<em>b</em>)RS</su<em>b</em>)), lower potential for recruitment with positive end-expiratory pressure (PEEP) contrasting with severe shunt fraction. The purpose of the study was to systematically assess respiratory mechanics and recruita<em>b</em>ility in SARS-Cov-2-associated ARDS.</p><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>Gas exchanges, C<su<em>b</em>)RS</su<em>b</em>) and hemodynamics were assessed at 2 levels of PEEP (15 cmH<su<em>b</em>)2</su<em>b</em>)O and 5 cmH<su<em>b</em>)2</su<em>b</em>)O) within 36 h (day1) and from 4 to 6 days (day 5) after intu<em>b</em>ation. The recruited volume was computed as the difference <em>b</em>etween the volume expired from PEEP 15 to 5 cmH<su<em>b</em>)2</su<em>b</em>)O and the volume predicted <em>b</em>y compliance at PEEP 5 cmH<su<em>b</em>)2</su<em>b</em>)O (or a<em>b</em>ove airway opening pressure). The recruitment-to-inflation (R/I) ratio (i.e. the ratio <em>b</em>etween the recruited lung compliance and C<su<em>b</em>)RS</su<em>b</em>) at PEEP 5 cmH<su<em>b</em>)2</su<em>b</em>)O) was used to assess lung recruita<em>b</em>ility. A R/I ratio value higher than or equal to 0.5 was used to define highly recruita<em>b</em>le patients.</p><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The R/I ratio was calculated in 25 of the 26 enrolled patients at day 1 and in 15 patients at day 5. At day 1, 16 (64%) were considered as highly recruita<em>b</em>le (R/I ratio median [interquartile range] 0.7 [0.55-0.94]) and 9 (36%) were considered as poorly recruita<em>b</em>le (R/I ratio 0.41 [0.31-0.48]). The PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) ratio at PEEP 15 cmH<su<em>b</em>)2</su<em>b</em>)O was higher compared to PEEP 5 cmH<su<em>b</em>)2</su<em>b</em>)O only in highly recruita<em>b</em>le patients (173 [139-236] vs 135 [89-167] mmHg; p &<em>lt</em>; 0.01). Neither PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) or C<su<em>b</em>)RS</su<em>b</em>) measured at PEEP 15 cmH<su<em>b</em>)2</su<em>b</em>)O or at PEEP 5 cmH<su<em>b</em>)2</su<em>b</em>)O nor changes in PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) or C<su<em>b</em>)RS</su<em>b</em>) in response to PEEP changes allowed to identify highly or poorly recruita<em>b</em>le patients.</p><A<em>b</em>stractText>In this series of 25 patients with SARS-Cov-2 associated ARDS, 64% were considered as highly recruita<em>b</em>le and only 36% as poorly recruita<em>b</em>le <em>b</em>ased on the R/I ratio performed on the day of intu<em>b</em>ation. This o<em>b</em>servation suggests that a systematic R/I ratio assessment may help to guide initial PEEP titration to limit harmful effect of unnecessary high PEEP in the context of Covid-19 crisis.</A<em>b</em>stractText>

Although studies indicate LIGHT (lymphotoxin (<em>LT</em>)-like, exhibits inducible expression and competes with HSV glycoprotein D for herpes virus entry mediator (HVEM), a receptor expressed by T lymphocytes) enhances inflammation and T cell-mediated immunity, the mechanisms involved in this process remain obscure. In this study, we assessed the role of LIGHT in IL-12 production and development of CD4(+) Th cells type one (Th1) in vivo. Bone marrow-derived dendritic cells from LIGHT(-/-) mice were severely impaired in IL-12p40 production following IFN-gamma and LPS stimulation in vitro. Furthermore, blockade of LIGHT in vitro and in vivo with HVEM-Ig and <em>LT</em> <em>beta</em> receptor (<em>LT</em><em>beta</em>R)-Ig leads to impaired IL-12 production and defective polyclonal and Ag-specific IFN-gamma production in vivo. In an infection model, injection of HVEM-Ig or <em>LT</em><em>beta</em>R-Ig into the usually resistant C57BL/6 mice results in defective IL-12 and IFN-gamma production and severe susceptibility to Leishmania major that was reversed by rIL-12 treatment. This striking susceptibility to L. major in mice injected with HVEM-Ig or <em>LT</em><em>beta</em>R-Ig was also reproduced in LIGHT(-/-) ->> RAG1(-/-) chimeric mice. In contrast, L. major-infected <em>LT</em><em>beta</em>(-/-) mice do not develop acute disease, suggesting that the effect of <em>LT</em><em>beta</em>R-Ig is not due to blockade of membrane <em>LT</em> (<em>LT</em>alpha1<em>beta</em>2) signaling. Collectively, our data show that LIGHT plays a critical role for optimal IL-12 production by DC and the development of IFN-gamma-producing CD4(+) Th1 cells and its blockade results in severe susceptibility to Leishmania major.

OBJECTIVE

Previous laboratory testing has identified the importance of upper-body aerobic and anaerobic power to cross-country skiing performance. The purpose of this investigation was to extend laboratory research into a field setting to identify predictors of performance through ski-specific testing.

METHODS

Thirteen male collegiate skiers performed three field-testing sessions on roller skis to establish lactate threshold (LT) and ski economy (ECON) and maximal oxygen uptake (SK VO(2max)) and a 1-km double-pole time trial (UBTT) to determine peak upper-body oxygen uptake (UB VO(2)). As a measure of skiing performance, the subjects performed a 10-km skating time trial (TT) and were ranked according to competitive season performance (RANK).

RESULTS

Significant correlations (P < 0.05) were found between SK VO(2max), LT VO(2), UB VO(2), and RANK (r = -0.66 to -0.84) and TT time (r = -0.74 to -0.79), as well as ECON to RANK (r = 0.57) and TT time (r = 0.68). Time to complete the UBTT (UB time) exhibited the strongest correlation to both RANK (r = 0.95) and TT time (r = 0.92). Multiple regression analyses revealed that UB time was the best predictor of RANK and TT time, as demonstrated by the significant beta values (0.77, P < 0.001, and 0.79, P < 0.001, respectively). The importance of the UB component was further seen in that UB time was still the best predictor of performance when the subjects were divided into two distinct groups of greater and lesser competitive ability.

CONCLUSIONS

These findings identify the importance of the upper body component to cross-country skiing performance, suggesting a need to focus on upper-body conditioning within a well-rounded endurance training program. Additionally, the UBTT exhibits potential as a simple field test to predict cross-country skiing performance over more sophisticated and costly laboratory and field testing.

Asthma is a chronic inflammatory disease that is associated with widespread but variable airflow obstruction. The mechanisms that lead to airflow obstruction in asthma are bronchoconstriction, mucosal edema, increased secretion of mucus, and an inflammatory-cell infiltrate that is rich in eosinophils. Leukotrienes (LTs) B4, C4, D4, and E4 have been shown experimentally to play a role in each of these inflammatory mechanisms and to mimic the pathologic changes seen in asthma. Inhaled LTC4 and LTD4 are the most potent bronchoconstrictors yet studied in human subjects. LTC4 and LTD4 also may cause migration of inflammatory cells into the asthmatic airway. LTs are derived from the 5-lipoxygenase (5-LO) pathway of arachidonic acid metabolism, and increased production of LTs has been demonstrated in patients who have asthma. Leukotriene receptor antagonists and specific inhibitors of the 5-LO pathway hold great promise as new therapies to treat asthma. Because LTC4, LTD4, and LTE4 appear to interact with a common LTD4 receptor, selective LTD4 receptor antagonists (eg, pranlukast [SB205312/ONO-1078], zafirlukast [ICI 204,219], MK-571, and MK-679), as well as zileuton (A-64077, a direct inhibitor of 5-LO) have been developed as antiasthma agents. Clinical and experimental studies have demonstrated the efficacy of these compounds in reducing not only the symptoms of asthma, but use of beta 2-agonists and bronchoconstriction induced by exposure to allergens, exercise, aspirin, and cold air.

(<em>b</em>)Background:</<em>b</em>) Checkpoint inhi<em>b</em>itors plus platinum-<em>b</em>ased chemotherapy have shown superiority compared to chemotherapy alone as first-line therapy in advanced non-small cell lung carcinoma (NSCLC). To evaluate the relative <em>b</em>enefit in term of Overall Survival (OS) and Progression-free Survival (PFS) of checkpoint inhi<em>b</em>itors plus chemotherapy vs. chemotherapy alone, overall and in su<em>b</em>groups defined <em>b</em>y PDL1 expression we have performed a meta-analysis. (<em>b</em>)Data Sources:</<em>b</em>) This meta-analysis searched Pu<em>b</em>Med and checked references of the selected English language articles to identify further eligi<em>b</em>le trials. Data collection for this study took place from Octo<em>b</em>er 1 to Octo<em>b</em>er 24, 2018. (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) In total, 8 trials involving 4,646 patients with advanced NSCLC, 3.314 (71%) and 1.332 (29%) with a non-squamous and squamous histology, respectively, were included in this meta-analysis. Four trials used atezolizuma<em>b</em>, 3 pem<em>b</em>rolizuma<em>b</em>, and 1 nivoluma<em>b</em>, accounting for 2.985 (64%), 1.298 (28%), and 363 (8%) of patients, respectively. The patients were randomized to receive first-line chemotherapy plus a checkpoint inhi<em>b</em>itor vs. first-line chemotherapy, 2,978 patients for the OS endpoint and first-line chemotherapy plus a checkpoint inhi<em>b</em>itor vs. first-line chemotherapy, 1,740 patients in the PFS endpoint. Checkpoint inhi<em>b</em>itors plus chemotherapy were associated with prolonged OS, compared with chemotherapy in the ITT population (HR, 0.74; 95% CI, 0.64-0.87; <i>p</i> = 0.0002, with significant heterogeneity among trials). Nota<em>b</em>ly within the PDL1 low group (1-49) there was a significant heterogeneity (<i>p</i> = 0.06) <em>b</em>etween type of drug and efficacy: the com<em>b</em>ination of chemotherapy plus pem<em>b</em>rolizuma<em>b</em> showed an OS <em>b</em>enefit (HR, 0.56; 95% CI, 0.40-0.78; <i>P</i> &<em>lt</em>; 0.00007) unlike the atezolizuma<em>b</em> <em>b</em>ack<em>b</em>one trials (HR, 0.92; 95% CI, 0.62-1.37; <i>P</i> &<em>lt</em>; 0.69). However, checkpoint inhi<em>b</em>itors plus chemotherapy were associated with prolonged PFS in the ITT (HR, 0.61; 95% CI, 0.56-0.66; <i>P</i> &<em>lt</em>; 0.00001) and across PDL1 su<em>b</em>groups. (<em>b</em>)Conclusion and Relevance:</<em>b</em>) Checkpoint inhi<em>b</em>itors plus chemotherapy compared with chemotherapy, are associated with significantly prolonged OS and PFS in first-line therapy in NSCLC. In the low PDL1 su<em>b</em>groups the <em>b</em>enefit was statistically significant only in the pem<em>b</em>rolizuma<em>b</em> <em>b</em>ack<em>b</em>one trials. The findings of this meta-analysis could assist in the design and interpretation of future trials and in economic analyses.

The anthrax protective antigen (PA) is the receptor-binding subunit common to lethal toxin (LT) and edema toxin (ET), which are responsible for the high mortality rates associated with inhalational Bacillus anthracis infection. Although recombinant PA (rPA) is likely to be an important constituent of any future anthrax vaccine, evaluation of the efficacies of the various candidate rPA vaccines is currently difficult, because the specific B-cell epitopes involved in toxin neutralization have not been completely defined. In this study, we describe the identification and characterization of two murine monoclonal immunoglobulin G1 antibodies (MAbs), 1-F1 and 2-BBLT. 2-BBBLT in vitro, although it only partially inhibited PA binding to its receptor. Mice passively administered 1-F1 or 2-BLT. These results advance our fundamental understanding of the mechanisms by which antibodies neutralize anthrax toxin and may have future application in the evaluation of candidate rPA vaccines.

To explore whether plasma circular RNAs (circRNAs) can diagnose hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), microarray and qPCR were used to identify plasma circRNAs that were increased in HBV-related HCC patients compared to controls (including healthy controls, chronic hepatitis B and HBV-related liver cirrhosis). A logistic regression model was constructed using a training set (n = 313) and then validated using another two independent sets (n = 306 and 526, respectively). Area under the receiver operating characteristic curve (AUC) was used to evaluate diagnostic accuracy. We identified a plasma circRNA panel (CircPanel) containing three circRNAs (hsa_circ_0000976, hsa_circ_0007750 and hsa_circ_0139897) that could detect HCC. CircPanel showed a higher accuracy than AFP (alpha-fetoprotein) to distinguish individuals with HCC from controls in all three sets (AUC, 0.863 [95% confidence interval, CI: 0.819-0.907] vs. 0.790 [0.738-0.842], p = 0.036 in training set; 0.843 [0.796-0.890] vs. 0.747 [0.691-0.804], p = 0.011 in validation set 1 and 0.864 [0.830-0.898] vs. 0.769 [0.728-0.810], p &lt; 0.001 in validation set 2). CircPanel also performed well in detecting Small-HCC (solitary, ≤3 cm), AFP-negative HCC and AFP-negative Small-HCC.

A<em>lt</em>ernative splicing (AS) has emerged as a key event in tumor development and microenvironment formation. However, comprehensive analysis of AS and its clinical significance in head and neck squamous cell carcinoma (HNSC) is urgently required. (<em>b</em>)Methods:</<em>b</em>) Genome-wide profiling of AS events using RNA-Seq data from The Cancer Genome Atlas (TCGA) program was performed in a cohort of 464 patients with HNSC. Cancer-associated AS events (CASEs) were identified <em>b</em>etween paired HNSC and adjacent normal tissues and evaluated in functional enrichment analysis. Splicing networks and prognostic models were constructed using <em>b</em>ioinformatics tools. Unsupervised clustering of the CASEs identified was conducted and associations with clinical, molecular and immune features were analyzed. (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) We detected a total of 32,309 AS events and identified 473 CASEs in HNSC; among these, 91 were validated in an independent cohort (n = 15). Functional protein domains were frequently a<em>lt</em>ered, especially <em>b</em>y CASEs affecting cancer drivers, such as PCSK5. CASE parent genes were significantly enriched in pathways related to HNSC and the tumor immune microenvironment, such as the viral carcinogenesis (FDR &<em>lt</em>; 0.001), Human Papillomavirus infection (FDR &<em>lt</em>; 0.001), chemokine (FDR &<em>lt</em>; 0.001) and T cell receptor (FDR &<em>lt</em>; 0.001) signaling pathways. CASEs enriched in immune-related pathways were closely associated with immune cell infi<em>lt</em>ration and cytolytic activity. AS regulatory networks suggested a significant association <em>b</em>etween splicing factor (SF) expression and CASEs and might <em>b</em>e regulated <em>b</em>y SF methylation. Eighteen CASEs were identified as independent prognostic factors for overall and disease-free survival. Unsupervised clustering analysis revealed distinct correlations <em>b</em>etween AS-<em>b</em>ased clusters and prognosis, molecular characteristics and immune features. Immunogenic features and immune su<em>b</em>groups cooperatively depict the immune features of AS-<em>b</em>ased clusters. (<em>b</em>)Conclusion:</<em>b</em>) This comprehensive genome-wide analysis of the AS landscape in HNSC revealed novel AS events related to carcinogenesis and immune microenvironment, with implications for prognosis and therapeutic responses.

The aim of the present study was to compare powdered rose hip with and without fruits (Rosae pseudofructus cum/sine fructibus, Rosa canina L., Rosaceae) with regard to their phytochemical profile and their in vitro anti-inflammatory and radical-scavenging properties. The two powders were subsequently extracted with solvents of increasing polarity and tested for inhibition of cyclooxygenase (COX-1, COX-2) and of 5-LOX-mediated leukotriene B(4) (LTB(4)) formation as well as for DPPH-radical-scavenging capacity. While the water and methanol extracts were inactive in the COX-1, COX-2 and LTB(4) inhibition assays, the n-hexane and the dichloromethane extracts inhibited all three enzymes. In the active extracts, the triterpenoic acids ursolic acid, oleanolic acid and betulinic acid were identified, although only in minute amounts. Furthermore, oleic, linoleic and alpha-linolenic acid were identified apart from several saturated fatty acids. Even though unsaturated fatty acids are known to be good inhibitors of COX-1, COX-2 and LT formation, no clear correlation between their concentration in the extracts and their activity was found. We suggest that other, yet unidentified, lipophilic constituents might play a more important role for the observed in vitro inhibitory activity on arachidonic acid metabolism. Some of the extracts also showed considerable DPPH radical scavenging activity, the methanolic extracts being most potent. The radical scavenging activity of the extracts correlated very well with their total phenolic content, while ascorbic acid contributes only little to the radical-scavenging activity due to its low concentration present in the extracts. In summary, extracts derived from powdered rose hip without fruits were more effective in all assays carried out compared with extracts derived from powdered rose hip with fruits.

S-Sulfocysteine synthase was isolated from Salmonella typhimurium LT-2 to homogeneous form with polyacrylamide gel electrophoresis. The molecular weight of this enzyme was determined to be ca. 55,000. The enzyme consisted of two identically sized subunits, and it contained one pyridoxal phosphate per subunit. The enzyme catalyzed the biosynthesis of cysteine or S-methylcysteine from sulfide or methanethiol and O-acetylserine, respectively, in addition to the formation of S-sulfocysteine from thiosulfate and O-acetylserine. The enzyme is identical to cysteine synthase B. The intracellular level of this enzyme was regulated by lesser extents of the same factors as those effective for cysteine synthase A.

Perinatal exposure to excess iodine can lead to transient hypothyroidism in the newborn. In Japan, large quantities of iodine-rich seaweed such as kombu (Laminaria japonica) are consumed. However, effects of iodine from food consumed during the perinatal period are unknown. The concentration of iodine in serum, urine, and breast milk in addition to thyrotropin (TSH), free thyroxine (FT(4)), and thyroglobulin was measured in 34 infants who were positive at congenital hypothyroidism screening. Based on the concentration of iodine in the urine, 15 infants were diagnosed with hyperthyrotropinemia caused by the excess ingestion of iodine by their mothers during their pregnancy. According to serum iodine concentrations, these infants were classified into group A (over 17 microg/dL) and group B (under 17 microg/dL) of serum iodine. During their pregnancies these mothers consumed kombu, other seaweeds, and instant kombu soups containing a high level of iodine. It was calculated that the mothers of group A infants ingested approximately 2300-3200 microg of iodine, and the mothers of group B infants approximately 820-1400 microg of iodine per day during their pregnancies. Twelve of 15 infants have required levo-thyroxine (LT(4)) because hypothyroxinemia or persistent hyperthyrotropinemia was present. In addition, consumption of iodine by the postnatal child and susceptibility to the inhibitory effect of iodine may contribute in part to the persistent hyperthyrotropinemia. We propose that hyperthyrotropinemia related to excessive iodine ingestion by the mother during pregnancy in some cases may not be transient.

<A<em>b</em>stractText>To determine the predictive and prognostic value of the consensus molecular su<em>b</em>types (CMSs) of colorectal cancer (CRC) that represent a merging of gene expression-<em>b</em>ased features largely in primary tumors from six independent classification systems and provide a framework for capturing the intrinsic heterogeneity of CRC in patients enrolled in CALGB/SWOG 80405.</A<em>b</em>stractText><A<em>b</em>stractText>CALGB/SWOG 80405 is a phase III trial that compared the addition of <em>b</em>evacizuma<em>b</em> or cetuxima<em>b</em> to infusional fluorouracil, leucovorin, and oxaliplatin or fluorouracil, leucovorin, and irinotecan as first-line treatment of advanced CRC. We characterized the CMS classification using a novel NanoString gene expression panel on primary CRCs from 581 patients enrolled in this study to assess the prognostic and predictive value of CMSs in these patients.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The CMSs are highly prognostic for overall survival (OS; <i>P</i> &<em>lt</em>; .001) and progression-free survival (PFS; <i>P</i> &<em>lt</em>; .001). Furthermore, CMSs were predictive for <em>b</em>oth OS (<i>P</i> for interaction &<em>lt</em>; .001) and PFS (<i>P</i> for interaction = .0032). In the CMS1 cohort, patients treated with <em>b</em>evacizuma<em>b</em> had a significantly longer OS than those treated with cetuxima<em>b</em> (<i>P</i> &<em>lt</em>; .001). In the CMS2 cohort, patients treated with cetuxima<em>b</em> had a significantly longer OS than patients treated with <em>b</em>evacizuma<em>b</em> (<i>P</i> = .0046).</p><A<em>b</em>stractText>These findings highlight the possi<em>b</em>le clinical utility of CMSs and suggests that refinement of the CMS classification may provide a path toward identifying patients with metastatic CRC who are most likely to <em>b</em>enefit from specific targeted therapy as part of the initial treatment.</A<em>b</em>stractText>

Macroautophagy/autophagy is a degradative process essential for various cellular processes. We previously demonstrated that autophagy-deficiency causes myo<em>b</em>last apoptosis and impairs myotu<em>b</em>e formation. In this study, we continued this work with particular emphasis on mitochondrial remodelling and stress/apoptotic signaling. We found increased (p &<em>lt</em>; 0.05) autophagic (e.g., a<em>lt</em>ered LC3<em>B</em> levels, increased ATG7, decreased SQSTM1) and mitophagic (e.g., <em>B</em>NIP3 upregulation, mitochondrial localized GFP-LC3 puncta, and elevated mitochondrial LC3<em>B</em>-II) signaling during myo<em>b</em>last differentiation. shRNA-mediated knockdown of ATG7 (sh<i>Atg7</i>) decreased these autophagic and mitophagic responses, while increasing CASP3 activity and ANXA5/annexin V staining in differentiating myo<em>b</em>lasts; u<em>lt</em>imately resu<em>lt</em>ing in dramatically impaired myogenesis. Further confirming the importance of mitophagy in these responses, CRISPR-Cas9-mediated knockout of <i><em>B</em>nip3</i> (<i><em>b</em>nip3^-/-</i>) resu<em>lt</em>ed in increased CASP3 activity and DNA fragmentation as well as impaired myo<em>b</em>last differentiation. In addition, sh<i>Atg7</i> myo<em>b</em>lasts displayed greater endoplasmic reticulum (e.g., increased CAPN activity and HSPA) and mitochondrial (e.g., mPTP formation, reduced mitochondrial mem<em>b</em>rane potential, elevated mitochondrial 4-HNE) stress. sh<i>Atg7</i> and <i><em>b</em>nip3</i>^-/- myo<em>b</em>lasts also displayed a<em>lt</em>ered mitochondria-associated signaling (e.g., PPARGC1A, DNM1L, OPA1) and protein content (e.g., SLC25A4, VDAC1, CYCS). Moreover, sh<i>Atg7</i> myo<em>b</em>lasts displayed CYCS and AIFM1 release from mitochondria, and CASP9 activation. Similarly, <i><em>b</em>nip3</i>^-/- myo<em>b</em>lasts had significantly higher CASP9 activation during differentiation. Importantly, administration of a chemical inhi<em>b</em>itor of CASP9 (Ac-LEHD-CHO) or dominant-negative CASP9 (ad-DNCASP9) partially recovered differentiation and myogenesis in sh<i>Atg7</i> myo<em>b</em>lasts. Together, these data demonstrate an essential role for autophagy in protecting myo<em>b</em>lasts from mitochondrial oxidative stress and apoptotic signaling during differentiation, as well as in the regulation of mitochondrial network remodelling and myogenesis. (<em>b</em>)A<em>b</em><em>b</em>reviations</<em>b</em>): 3MA: 3-methyladenine; 4-HNE: 4-hydroxynonenal; ACT: actin; AIFM1/AIF: apoptosis-inducing factor, mitochondrion-associated 1; ANXA5: annexin V; ATG7: autophagy related 7; AU: ar<em>b</em>itrary units; <em>B</em>AX: <em>B</em>CL2-associated X protein; <em>B</em>CL2: <em>B</em> cell leukemia/lymphoma 2; <em>B</em>ECN1: <em>b</em>eclin 1, autophagy related; <em>B</em>NIP3: <em>B</em>CL2/adenovirus E1<em>B</em> interacting protein 3; CAPN: calpain; CASP: caspase; CASP3: caspase 3; CASP8: caspase 8; CASP9: caspase 9; CASP12: caspase 12; CAT: catalase; CQ: chloroquine; CYCS: cytochrome c, somatic; DCF; 2',7'-dichlorofluorescein; DNM1L/DRP1: dynamin 1-like; DM: differentiation media; DMEM: Dul<em>b</em>ecco's modified Eagle's medium; ER: endoplasmic reticulum; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; GM: growth media; p-H2AFX: phosphorylated H2A histone family, mem<em>b</em>er X; H2<em>B</em>FM: H2<em>B</em> histone family, mem<em>b</em>er M; H<em>B</em>SS: Hanks <em>b</em>alanced sa<em>lt</em> solution; HSPA/HSP70: heat shock protein family A; JC-1: tetraethyl<em>b</em>enzimidazolylcar<em>b</em>ocyanine iodide; MAP1LC3<em>B</em>/LC3<em>B</em>: microtu<em>b</em>ule-associated protein 1 light chain 3 <em>beta</em>; mPTP: mitochondrial permea<em>b</em>ility transition pore; MYH: myosin heavy chain; MYOG: myogenin; OPA1: OPA1, mitochondrial dynamin like GTPase; PI: propidium iodide; PINK1: PTEN induced putative kinase 1; PPARGC1A/PGC1α: peroxisome proliferative activated receptor, gamma, coactivator 1 alpha; ROS: reactive oxygen species; SLC25A4/ANT1: solute carrier family 25 (mitochondrial carrier, adenine nucleotide translocator), mem<em>b</em>er 4; SOD1: superoxide dismutase 1, solu<em>b</em>le; SOD2: superoxide dismutase 2, mitochondrial; SQSTM1/p62: sequestosome 1; VDAC1: vo<em>lt</em>age-dependent anion channel 1.

Objective: Mindfulness disposition is associated with various psychological factors and prevents emotional distress in chronic diseases. In the present study, we analyzed the key role of mindfulness dispositions in protecting the individual against psychological distress consequent to COVID-19 social distancing and quarantining.

Methods: An online survey was launched on March 13, 2020, with 6,412 responses by April 6, 2020. Socio-demographic information, exposure to the pandemic, and quarantining were assessed together with psychological distress and mindfulness disposition. Multivariate linear regression analysis was performed to study the influence of predictive factors on psychological distress and quality of life in Italian responders during the early days of lockdown. Pearson correlations were calculated to study the relationship between mindfulness and psychiatric symptoms.

Results: Multivariate linear regression run on socio-demographics, COVID-19-related variables, and mindfulness disposition as moderators of overall psychological distress showed that mindfulness was the best predictor of psychological distress (β = -0.504; p &lt; 0.0001). High negative correlations were found between mindfulness disposition and the overall Global Severity Index (r = -0.637; p &lt; 0.0001), while moderate to high associations were found between mindfulness and all SCL-90 sub-scales.

Discussion: Findings showed that high dispositional mindfulness enhances well-being and helps in dealing with stressful situations such as the COVID-19 pandemic. Mindfulness-based mental training could represent an effective intervention to stem post-traumatic psychopathological beginnings and prevent the onset of chronic mental disorders.

Keywords: COVID-19; MAAS; SCL-90; adjustment; meditation; mindfulness; pandemic; psychological distress.

Immune checkpoint inhibition suggests promising progress for the treatment of advanced hepatocellular carcinoma (HCC). However, the underlying cellular mechanisms remain unclear because liver cancer cells apparently do not upregulate inhibitory checkpoint molecules. Here, we analysed whether regulatory T cells (Tregs) can alternatively trigger checkpoint inhibition pathways in HCC. Using flow cytometry we analysed expression of checkpoint molecules (PD-1, PD-L1, CTLA-4, GITR, Tim-3) on peripheral CD4⁺CD25⁺Foxp3⁺ Tregs and their secretion of inhibitory mediators (IL-10, IL-35, TGF-beta, galectin-9) in 116 individuals (50 patients with HCC, 41 non-tumour bearing liver disease controls, 25 healthy controls). Functional activity of Tregs on T effector cells (IFN-gamma production, cytotoxicity) was characterized in vitro using a lectin-dependent cellular cytotoxicity (LDCC) assay against checkpoint inhibitor-negative P815 target cells. Unlike liver patients without malignancy and healthy controls, the frequency of checkpoint inhibitor-positive Tregs inversely correlated to age of patients with HCC (PD-L1, p = 0.0080; CTLA-4, p = 0.0029) and corresponded to enhanced numbers of Tregs producing IL-10 and IL-35 (p &lt; 0.05 each). Tregs inhibited IFN-gamma secretion and cytotoxicity of CD8⁺ T cells when added to LDCC against P815 cells. Treg-induced inhibition of IFN-gamma secretion could be partially blocked by neutralizing PD-1 and PD-L1 antibodies specifically in HCC patients. In HCC peripheral Tregs upregulate checkpoint inhibitors and contribute to systemic immune dysfunction and antitumoural activity by several inhibitory pathways, presumably facilitating tumour development at young age. Blocking PD-L1/PD-1 interactions in vitro selectively interfered with inhibitory Treg -T effector cell interactions in the patients with HCC and resulted in improved antitumoural activity also against checkpoint inhibitor-negative tumour cells.

<A<em>b</em>stractText>To use the large dataset from the Tysa<em>b</em>ri Outreach: Unified Commitment to Hea<em>lt</em>h (TOUCH) program to compare progressive mu<em>lt</em>ifocal leukoencephalopathy (PML) risk with natalizuma<em>b</em> extended interval dosing (EID) vs standard interval dosing (SID) in patients with mu<em>lt</em>iple sclerosis (MS).</A<em>b</em>stractText><A<em>b</em>stractText>This retrospective cohort study included anti-JC virus anti<em>b</em>ody-positive patients (n = 35,521) in the TOUCH data<em>b</em>ase as of June 1, 2017. The effect of EID on PML risk was evaluated with 3 planned analyses using Kaplan-Meier methods stratified <em>b</em>y prior immunosuppressant use. Risk of PML was analyzed <em>b</em>y Cox regression adjusted for age, sex, prior immunosuppressants, time since natalizuma<em>b</em> initiation, and cumulative num<em>b</em>er of infusions.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>This study included 35,521 patients (primary analysis: 1,988 EID, 13,132 SID; secondary analysis: 3,331 EID, 15,424 SID; tertiary analysis: 815 EID, 23,168 SID). Mean average dosing intervals were 35.0 to 43.0 and 29.8 to 30.5 days for the EID and SID cohorts, respectively. Hazard ratios (95% confidence intervals) of PML risk for EID vs SID were 0.06 (0.01-0.22, <i>p</i> &<em>lt</em>; 0.001) and 0.12 (0.05-0.29, <i>p</i> &<em>lt</em>; 0.001) for the primary and secondary analyses, respectively. Relative risk reductions were 94% and 88% in favor of EID for the primary and secondary analyses, respectively. The tertiary analysis included no cases of PML with EID.</p><A<em>b</em>stractText>Natalizuma<em>b</em> EID is associated with clinically and statistically significantly lower PML risk than SID.</A<em>b</em>stractText><A<em>b</em>stractText>This study provides Class III evidence that for patients with MS, natalizuma<em>b</em> EID is associated with a lower PML risk than SID.</A<em>b</em>stractText>

BACKGROUND

Some experimental trials have demonstrated that rapamycin (RAPA) is able to inhibit HIV-1 progression in three different ways: (1) reducing CCR5-gene transcription, (2) blocking interleukin-2 intracellular secondary messenger (mammalian target of rapamycin), and (3) up-regulating the beta-chemokine macrophage inflammatory protein (MIP; MIP-1alpha and MIP-1beta). We present the preliminary results of a prospective nonrandomized trial concerning the first HIV patient series receiving RAPA monotherapy after liver transplantation (LT).

METHODS

Since June 2003, 14 HIV patients have received cadaveric donor LT due to end-stage liver disease (ESLD) associated or not associated with hepatocellular carcinoma, scored by the model for ESLD system. Patients were assessed using the following criteria for HIV characterization: CD4 T-cell count more than 100/mL and HIV-RNA levels less than 50 copies/mL. Primary immunosuppression was based on calcineurin inhibitors (CI), whereas switch to RAPA monotherapy occurred in cases of CI complications or Kaposi's sarcoma.

RESULTS

Mean overall post-LT follow-up was 14.8 months (range: 0.5-52.6). Six of 14 patients were administered RAPA monotherapy. Mean preswitch period from CI to RAPA was 67 days (range: 10-225 days). Mean postswitch follow-up was 11.9 months (range: 2-31 months). All patients were affected by ESLD, which was associated with hepatocellular carcinoma in seven patients. ESLD occurred due to hepatitis C virus (HCV)-related hepatopathy for nine patients, hepatitis B virus-related hepatopathy for one patient, and hepatitis B virus-HCV hepatopathy for four patients. Significantly better control of HIV and HCV replication was found among patients taking RAPA monotherapy (P=0.0001 and 0.03, respectively).

CONCLUSIONS

After in vitro and in vivo experimental evidence of RAPA antiviral proprieties, to our knowledge, this is the first clinical report of several significant benefits in long-term immunosuppression maintenance and HIV-1 control among HIV positive patients who underwent LT.