Crude preparations of interferon (IFN)-gamma derived from human peripheral blood leukocyte (PBL) cultures induced with 12-O-tetra-decanoylphorbol-13-acetate (TPA) and phytohemagglutinin (PHA) were more cytotoxic to HeLa cells than partially purified nautral or highly purified recombinant human IFN-gamma preparations. Conditioned media from PBL cultures contained, in addition to IFN-gamma, a mixture of cytotoxins, including classic lymphocyte-derived lymphotoxin (LT), and a TPA-induced cytotoxic activity produced by the adherent cell population (presumably monocytes). These two types of cytotoxins, indistinguishable in the mouse L929 cell LT assay, could be differentiated by an antiserum prepared against LT derived from the B lymphoblastoid cell line RPMI 1788. This antiserum neutralized lymphocyte-derived classic LT but failed to neutralize the activity of the monocyte-derived cytotoxin. Processing of conditioned media by sequential chromatography on silicic acid, Con A-Sepharose, and DEAE-Sephacel failed to separate IFN-gamma from the LT activity. However, this procedure did remove the monocyte-derived cytotoxic activity present in the original starting material, leaving predominantly classic LT. This LT showed a slightly basic isoelectric point (pI 7.6) which partially overlapped the more basic pI range of IFN-gamma. The two lymphokine activities also could not be completely separated by fast protein liquid chromatography or molecular sieve chromatography. LT in these partially purified preparations was associated with a protein having an apparent molecular weight of 58,000 on gel filtration. This form dissociated partially into a 20,000 mol wt species after denaturation with 0.1% NaDodSO4. IFN-gamma could be selectively removed from preparations containing both IFN-gamma and LT with the aid of monoclonal antibody to IFN-gamma. The addition of purified LT to purified E. coli-derived recombinant human IFN-gamma resulted in a marked synergistic enhancement of cytotoxicity for HeLa cells.

<p><div>(<em>b</em>)BACKGROUND</<em>b</em>)</div>Previous studies suggest that prone positioning (PP) can increase PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) and reduce mortality in moderate to severe acute respiratory distress syndrome (ARDS). The aim of our study was to determine whether the early use of PP com<em>b</em>ined with non-invasive ventilation (NIV) or high-flow nasal cannula (HFNC) can avoid the need for intu<em>b</em>ation in moderate to severe ARDS patients.</p><A<em>b</em>stractText>This prospective o<em>b</em>servational cohort study was performed in two teaching hospitals. Non-intu<em>b</em>ated moderate to severe ARDS patients were included and were placed in PP with NIV or with HFNC. The efficacy in improving oxygenation with four support methods-HFNC, HFNC+PP, NIV, NIV+PP-were evaluated <em>b</em>y <em>b</em>lood gas analysis. The primary outcome was the rate of intu<em>b</em>ation.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>Between January 2018 and April 2019, 20 ARDS patients were enrolled. The main causes of ARDS were pneumonia due to influenza (9 cases, 45%) and other viruses (2 cases, 10%). Ten cases were moderate ARDS and 10 cases were severe. Eleven patients avoided intu<em>b</em>ation (success group), and 9 patients were intu<em>b</em>ated (failure group). All 7 patients with a PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) &<em>lt</em>; 100 mmHg on NIV required intu<em>b</em>ation. PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) in HFNC+PP were significantly higher in the success group than in the failure group (125 ± 41 mmHg vs 119 ± 19 mmHg, P = 0.043). PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) demonstrated an upward trend in patients with all four support strategies: HFNC &<em>lt</em>; HFNC+PP ≤ NIV &<em>lt</em>; NIV+PP. The average duration for PP was 2 h twice daily.</p><p><div>(<em>b</em>)CONCLUSIONS</<em>b</em>)</div>Early application of PP with HFNC, especially in patients with moderate ARDS and <em>b</em>aseline SpO<su<em>b</em>)2</su<em>b</em>) > 95%, may help avoid intu<em>b</em>ation. The PP was well tolerated, and the efficacy on PaO<su<em>b</em>)2</su<em>b</em>)/FiO<su<em>b</em>)2</su<em>b</em>) of the four support strategies was HFNC &<em>lt</em>; HFNC+PP ≤ NIV &<em>lt</em>; NIV+PP. Severe ARDS patients were not appropriate candidates for HFNC/NIV+PP.</p><A<em>b</em>stractText>ChiCTR, ChiCTR1900023564. Registered 1 June 2019 (retrospectively registered).</A<em>b</em>stractText>

Although several cytokines, including tumor necrosis factor (TNF), can promote the growth of dendritic cells (DCs) in vitro, the cytokines that naturally regulate DC development and function in vivo have not been well defined. Here, we report that membrane lymphotoxin (LT), instead of TNF, regulates the migration of DCs in the spleen. LTalpha(-/-) mice, lacking membrane LTalpha/beta and LTalpha(3), show markedly reduced numbers of DCs in the spleen. Unlike wild-type mice and TNF(-/-) mice that have densely clustered DCs in the T cell zone and around the marginal zone, splenic DCs in LTalpha(-/-) mice are randomly distributed. The reduced number of DCs in lymphoid tissues of LTalpha(-/-) mice is associated with an increased number of DCs in nonlymphoid tissues. The number of splenic DCs in LTalpha(-/-) mice is restored when additional LT-expressing cells are provided. Blocking membrane LTalpha/beta in wild-type mice markedly diminishes the accumulation of DCs in lymphoid tissues. These data suggest that membrane LT is an essential ligand for the presence of DCs in the spleen. Mice deficient in TNF receptor, which is the receptor for both soluble LTalpha(3) and TNF-alpha(3) trimers, have normal numbers of DCs. However, LTbetaR(-/-) mice show reduced numbers of DCs, similar to the mice lacking membrane LT alpha/beta. Taken together, these results support the notion that the signaling via LTbetaR by membrane LTalpha/beta is required for the presence of DCs in lymphoid tissues.

The NF-κB transcription factor regulates numerous immune responses but its contribution to interleukin-17 (IL-17) production by T cells is largely unknown. Here, we report that IL-17, but not interferon-γ (IFN-γ), production by γδ T cells required the NF-κB family members RelA and RelB as well as the lymphotoxin-β-receptor (LTβR). In contrast, LTβR-NF-κB signaling was not involved in the differentiation of conventional αβ Th17 cells. Impaired IL-17 production in RelA- or RelB-deficient T cells resulted in a diminished innate immune response to Escherichia coli infection. RelA controlled the expression of LT ligands in accessory thymocytes whereas RelB, acting downstream of LTβR, was required for the expression of RORγt and RORα4 transcription factors and the differentiation of thymic precursors into γδT17 cells. Thus, RelA and RelB within different thymocyte subpopulations cooperate in the regulation of IL-17 production by γδ T cells and contribute to the host's ability to fight bacterial infections.

(<em>b</em>)O<em>b</em>jective:</<em>b</em>) To evaluate the cardiovascular damage of patients with COVID-19, and determine the correlation of serum N-terminal pro <em>B</em>-type natriuretic peptide (NT-pro<em>B</em>NP) and cardiac troponin-I (cTnI) with the severity of COVID-19, and the impact of concomitant cardiovascular disease on severity of COVID-19 was also evaluated. (<em>b</em>)Methods:</<em>b</em>) A cross-sectional study was designed on 150 consecutive patients with COVID-19 in the fever clinic of Tongji Hospital in Wuhan from January to Fe<em>b</em>ruary in 2020, including 126 mild cases and 24 cases in critical care. <em>B</em>oth univariate and mu<em>lt</em>ivariate logistic regression were used to analyze the correlation of past medical history including hypertension, dia<em>b</em>etes and coronary heart disease (CHD) , as well as the levels of serum NT-pro<em>B</em>NP and cTnI to the disease severity of COVID-19 patients. (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) Age, hypersensitive C-reactive protein(hs-CRP) and serum creatinine levels of the patients were higher in critical care cases than in mild cases(all <i>P</i>&<em>lt</em>;0.05). Prevalence of male, elevated NT-pro<em>B</em>NP and cTnI, hypertension and coronary heart disease were significantly higher in critical cases care patients than in the mild cases(all <i>P</i>&<em>lt</em>;0.05). Univariate logistic regression analysis showed that age, male, elevated NT-pro<em>B</em>NP, elevated cTnI, elevated hs-CRP, elevated serum creatinine, hypertension, and CHD were significantly correlated with critical disease status(all <i>P</i>&<em>lt</em>;0.05). Mu<em>lt</em>ivariate logistic regression analysis showed that elevated cTnI(<i>OR</i>=26.909, 95%<i>CI</i> 4.086-177.226, <i>P</i>=0.001) and CHD (<i>OR</i>=16.609, 95%<i>CI</i> 2.288-120.577, <i>P</i>=0.005) were the independent risk factors of critical disease status. (<em>b</em>)Conclusions:</<em>b</em>) COVID-19 can significantly affect the heart function and lead to myocardial injury. The past medical history of CHD and increased level of cTnI are two independent determinants of clinical disease status in patients with COVID-19.

<A<em>b</em>stractText>To investigate whether diets differing in fat content a<em>lt</em>er the gut micro<em>b</em>iota and faecal meta<em>b</em>olomic profiles, and to determine their relationship with cardiometa<em>b</em>olic risk factors in hea<em>lt</em>hy adu<em>lt</em>s whose diet is in a transition from a traditional low-fat diet to a diet high in fat and reduced in car<em>b</em>ohydrate.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>In a 6-month randomised controlled-feeding trial, 217 hea<em>lt</em>hy young adu<em>lt</em>s (aged 18-35 years; <em>b</em>ody mass index &<em>lt</em>;28 kg/m²; 52% women) who completed the whole trial were included. All the foods were provided during the intervention period. The three isocaloric diets were: a lower-fat diet (fat 20% energy), a moderate-fat diet (fat 30% energy) and a higher-fat diet (fat 40% energy). The effects of the dietary interventions on the gut micro<em>b</em>iota, faecal meta<em>b</em>olomics and plasma inflammatory factors were investigated.</p><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>The lower-fat diet was associated with increased α-diversity assessed <em>b</em>y the Shannon index (p=0.03), increased a<em>b</em>undance of <i>Blautia</i> (p=0.007) and <i>Faecali<em>b</em>acterium</i> (p=0.04), whereas the higher-fat diet was associated with increased <i>Alistipes</i> (p=0.04), <i>Bacteroides</i> (p&<em>lt</em>;0.001) and decreased <i>Faecali<em>b</em>acterium</i> (p=0.04). The concentration of total short-chain fatty acids was significantly decreased in the higher-fat diet group in comparison with the other groups (p&<em>lt</em>;0.001). The cometa<em>b</em>olites p-cresol and indole, known to <em>b</em>e associated with host meta<em>b</em>olic disorders, were decreased in the lower-fat diet group. In addition, the higher-fat diet was associated with faecal enrichment in arachidonic acid and the lipopolysaccharide <em>b</em>iosynthesis pathway as well as elevated plasma proinflammatory factors after the intervention.</p><A<em>b</em>stractText>Higher-fat consumption <em>b</em>y hea<em>lt</em>hy young adu<em>lt</em>s whose diet is in a state of nutrition transition appeared to <em>b</em>e associated with unfavoura<em>b</em>le changes in gut micro<em>b</em>iota, faecal meta<em>b</em>olomic profiles and plasma proinflammatory factors, which might confer adverse consequences for long-term hea<em>lt</em>h outcomes.</A<em>b</em>stractText><A<em>b</em>stractText>NCT02355795; Resu<em>lt</em>s.</A<em>b</em>stractText>

(<em>b</em>)Background:</<em>b</em>) Recently discovered relationships <em>b</em>etween the gastrointestinal micro<em>b</em>iome and the <em>b</em>rain have implications for psychiatric disorders, including major depressive disorder (MDD). Bacterial transplantation from MDD patients to rodents produces depression-like <em>b</em>ehaviors. In humans, case-control studies have examined the gut micro<em>b</em>iome in hea<em>lt</em>hy and affected individuals. We systematically reviewed existing studies comparing gut micro<em>b</em>ial composition in MDD and hea<em>lt</em>hy volunteers. (<em>b</em>)Methods:</<em>b</em>) A Pu<em>b</em>Med literature search com<em>b</em>ined the terms "depression," "depressive disorder," "stool," "fecal," "gut," and "micro<em>b</em>iome" to identify human case-control studies that investigated relationships <em>b</em>etween MDD and micro<em>b</em>iota quantified from stool. We evaluated the resu<em>lt</em>ing studies, focusing on <em>b</em>acterial taxa that were different <em>b</em>etween MDD and hea<em>lt</em>hy controls. (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) Six eligi<em>b</em>le studies were found in which 50 taxa exhi<em>b</em>ited differences (<i>p</i> &<em>lt</em>; 0.05) <em>b</em>etween patients with MDD and controls. Patient characteristics and methodologies varied widely <em>b</em>etween studies. Five phyla-<i>Bacteroidetes, Firmicutes, Actino<em>b</em>acteria, Fuso<em>b</em>acteria</i>, and <i>Proto<em>b</em>acteria</i>-were represented; however, divergent resu<em>lt</em>s occurred across studies for all phyla. The largest num<em>b</em>er of differentiating taxa were within phylum <i>Firmicutes</i>, in which nine families and 12 genera differentiated the diagnostic groups. The majority of these families and genera were found to <em>b</em>e statistically different <em>b</em>etween the two groups in two identified studies. Family <i>Lachnospiraceae</i> differentiated the diagnostic groups in four studies (with an even split in directionality). Across all five phyla, nine genera were higher in MDD (<i>Anaerostipes, Blautia, Clostridium, Kle<em>b</em>siella, Lachnospiraceae incertae sedis, Para<em>b</em>acteroides, Parasutterella, Phascolarcto<em>b</em>acterium</i>, and <i>Streptococcus</i>), six were lower (<i>Bifido<em>b</em>acterium, Dialister, Escherichia/Shigella, Faecali<em>b</em>acterium</i>, and <i>Ruminococcus</i>), and six were divergent (<i>Alistipes, Bacteroides, Megamonas, Oscilli<em>b</em>acter, Prevotella</i>, and <i>Rose<em>b</em>uria</i>). We highlight mechanisms and products of <em>b</em>acterial meta<em>b</em>olism as they may relate to the etiology of depression. (<em>b</em>)Conclusions:</<em>b</em>) No consensus has emerged from existing human studies of depression and gut micro<em>b</em>iome concerning which <em>b</em>acterial taxa are most relevant to depression. This may in part <em>b</em>e due to differences in study design. Given that <em>b</em>acterial functions are conserved across taxonomic groups, we propose that studying micro<em>b</em>ial functioning may <em>b</em>e more productive than a purely taxonomic approach to understanding the gut micro<em>b</em>iome in depression.

<A<em>b</em>stractText>The coronavirus disease 2019 (COVID-19) is rapidly spreading in China and more than 30 countries over last two months. COVID-19 has mu<em>lt</em>iple characteristics distinct from other infectious diseases, including high infectivity during incu<em>b</em>ation, time delay <em>b</em>etween real dynamics and daily o<em>b</em>served num<em>b</em>er of confirmed cases, and the intervention effects of implemented quarantine and control measures.</A<em>b</em>stractText><A<em>b</em>stractText>We develop a Suscepti<em>b</em>le, Un-quanrantined infected, Quarantined infected, Confirmed infected (SUQC) model to characterize the dynamics of COVID-19 and explicitly parameterize the intervention effects of control measures, which is more suita<em>b</em>le for analysis than other existing epidemic models.</A<em>b</em>stractText><p><div>(<em>b</em>)Resu<em>lt</em>s</<em>b</em>)</div>The SUQC model is applied to the daily released data of the confirmed infections to analyze the out<em>b</em>reak of COVID-19 in Wuhan, Hu<em>b</em>ei (excluding Wuhan), China (excluding Hu<em>b</em>ei) and four first-tier cities of China. We found that, <em>b</em>efore January 30, 2020, all these regions except Beijing had a reproductive num<em>b</em>er <i>R</i> > 1, and after January 30, all regions had a reproductive num<em>b</em>er <i>R</i> &<em>lt</em>; 1, indicating that the quarantine and control measures are effective in preventing the spread of COVID-19. The confirmation rate of Wuhan estimated <em>b</em>y our model is 0.0643, su<em>b</em>stantially lower than that of Hu<em>b</em>ei excluding Wuhan (0.1914), and that of China excluding Hu<em>b</em>ei (0.2189), <em>b</em>ut it jumps to 0.3229 after Fe<em>b</em>ruary 12 when clinical evidence was adopted in new diagnosis guidelines. The num<em>b</em>er of unquarantined infected cases in Wuhan on Fe<em>b</em>ruary 12, 2020 is estimated to <em>b</em>e 3,509 and declines to 334 on Fe<em>b</em>ruary 21, 2020. After fitting the model with data as of Fe<em>b</em>ruary 21, 2020, we predict that the end time of COVID-19 in Wuhan and Hu<em>b</em>ei is around late March, around mid March for China excluding Hu<em>b</em>ei, and <em>b</em>efore early March 2020 for the four tier-one cities. A total of 80,511 individuals are estimated to <em>b</em>e infected in China, among which 49,510 are from Wuhan, 17,679 from Hu<em>b</em>ei (excluding Wuhan), and the rest 13,322 from other regions of China (excluding Hu<em>b</em>ei). Note that the estimates are from a deterministic ODE model and should <em>b</em>e interpreted with some uncertainty.</p><A<em>b</em>stractText>We suggest that rigorous quarantine and control measures should <em>b</em>e kept <em>b</em>efore early March in Beijing, Shanghai, Guangzhou and Shenzhen, and <em>b</em>efore late March in Hu<em>b</em>ei. The model can also <em>b</em>e useful to predict the trend of epidemic and provide quantitative guide for other countries at high risk of out<em>b</em>reak, such as South Korea, Japan, Italy and Iran.</A<em>b</em>stractText><A<em>b</em>stractText>The supplementary materials can <em>b</em>e found online with this article at 10.1007/s40484-020-0199-0.</A<em>b</em>stractText>

OBJECTIVE

Liver resection (LR) and liver transplantation (LT) are considered the only two potentially curative treatments for hepatocellular carcinoma (HCC). Recently, there has been an intense debate as to whether LR or LT is the optimal initial treatment for patients with Child A or B cirrhosis. The aim of this study was to compare the results of LR and LT in patients with HCC and with Child A or B cirrhosis in a single center over a 10-year period.

METHODS

Seventy-eight patients were treated with LT and 130 were treated with LR. We evaluated patient characteristics, short-term results such as hospital stay, postoperative complication, mortality, and long-term results such as overall and recurrence-free survival and recurrence.

RESULTS

The hospital stay of the LT group was significantly longer than that of the LR group (P < 0.001). The postoperative complication rate and the early operative mortality rate were similar between the two groups. The overall survival rate was higher after LT than it was after LR, but not to a statistically significant degree (P = 0.267). The recurrence-free survival rate was significantly higher after LT than it was after LR (P = 0.002). Within and beyond the Milan criteria, the overall survival rate was higher after LT than it was after LR, but not to a statistically significant degree. The recurrence-free survival rate was significantly higher after LT than it was after LR in the patients within Milan criteria (P < 0.001). HCC recurred more frequently after resection (51.5%) than it did after transplantation (29.5%) (P < 0.001), and HCC recurrence developed in the liver more frequently after LR than it did after LT (P = 0.002). However, after recurrence, LR had better survival than LT did, but not to a statistically significant degree (P = 0.177).

CONCLUSIONS

LT should be considered as the primary treatment in patients with HCC within the Milan criteria. LR is recommended for patients with HCC beyond the Milan criteria. The LT group showed a significantly lower recurrence rate than the LR group. However, in the case of recurrence, the LT group showed a poorer long-term outcome than the LR group.

Transmissible spongiform encephalopathies (TSEs) may be acquired peripherally, in which case infectivity usually accumulates in lymphoid tissues before dissemination to the nervous system. Studies of mouse scrapie models have shown that mature follicular dendritic cells (FDCs), expressing the host prion protein (PrP(c)), are critical for replication of infection in lymphoid tissues and subsequent neuroinvasion. Since FDCs require lymphotoxin signals from B lymphocytes to maintain their differentiated state, blockade of this stimulation with a lymphotoxin beta receptor-immunoglobulin fusion protein (LT beta R-Ig) leads to their temporary dedifferentiation. Here, a single treatment with LT beta R-Ig before intraperitoneal scrapie inoculation blocked the early accumulation of infectivity and disease-specific PrP (PrP(Sc)) within the spleen and substantially reduced disease susceptibility. These effects coincided with an absence of FDCs in the spleen for ca. 28 days after treatment. Although the period of FDC dedifferentiation was extended to at least 49 days by consecutive LT beta R-Ig treatments, this had little added protective benefit after injection with a moderate dose of scrapie. We also demonstrate that mature FDCs are critical for the transmission of scrapie from the gastrointestinal tract. Treatment with LT beta R-Ig before oral scrapie inoculation blocked PrP(Sc) accumulation in Peyer's patches and mesenteric lymph nodes and prevented neuroinvasion. However, treatment 14 days after oral inoculation did not affect survival time or susceptibility, suggesting that infectivity may have already spread to the peripheral nervous system. Although manipulation of FDCs may offer a potential approach for early intervention in peripherally acquired TSEs, these data suggest that the duration of the treatment window may vary widely depending on the route of exposure.

Development of a small animal model for the in vivo study of human immunity and infectious disease remains an important goal, particularly for investigations of HIV vaccine development. NOD/Lt mice homozygous for the severe combined immunodeficiency (Prkdcscid) mutation readily support engraftment with high levels of human hematolymphoid cells. However, NOD/LtSz-scid mice are highly radiosensitive, have short life spans, and a small number develop functional lymphocytes with age. To overcome these limitations, we have backcrossed the null allele of the recombination-activating gene (Rag1) for 10 generations onto the NOD/LtSz strain background. Mice deficient in RAG1 activity are unable to initiate V(D)J recombination in Ig and TCR genes and lack functional T and B lymphocytes. NOD/LtSz-Rag1null mice have an increased mean life span compared with NOD/LtSz-scid mice due to a later onset of lymphoma development, are radioresistant, and lack serum Ig throughout life. NOD/LtSz-Rag1null mice were devoid of mature T or B cells. Cytotoxic assays demonstrated low NK cell activity. NOD/LtSz-Rag1null mice supported high levels of engraftment with human lymphoid cells and human hemopoietic stem cells. The engrafted human T cells were readily infected with HIV. Finally, NOD/LtSz-Rag1null recipients of adoptively transferred spleen cells from diabetic NOD/Lt+/+ mice rapidly developed diabetes. These data demonstrate the advantages of NOD/LtSz-Rag1null mice as a radiation and lymphoma-resistant model for long-term analyses of engrafted human hematolymphoid cells or diabetogenic NOD lymphoid cells.

It remains unclear what determines the subcellular localization of hepatitis B virus (HBV) core protein (HBc) and particles. To address this fundamental issue, we have identified four distinct HBc localization signals in the arginine rich domain (ARD) of HBc, using immunofluorescence confocal microscopy and fractionation/Western blot analysis. ARD consists of four tight clustering arginine-rich subdomains. ARD-I and ARD-III are associated with two co-dependent nuclear localization signals (NLS), while ARD-II and ARD-IV behave like two independent nuclear export signals (NES). This conclusion is based on five independent lines of experimental evidence: i) Using an HBV replication system in hepatoma cells, we demonstrated in a double-blind manner that only the HBc of mutant ARD-II+IV, among a total of 15 ARD mutants, can predominantly localize to the nucleus. ii) These results were confirmed using a chimera reporter system by placing mutant or wild type HBc trafficking signals in the heterologous context of SV40 large T antigen (LT). iii) By a heterokaryon or homokaryon analysis, the fusion protein of SV40 LT-HBc ARD appeared to transport from nuclei of transfected donor cells to nuclei of recipient cells, suggesting the existence of an NES in HBc ARD. This putative NES is leptomycin B resistant. iv) We demonstrated by co-immunoprecipitation that HBc ARD can physically interact with a cellular factor TAP/NXF1 (Tip-associated protein/nuclear export factor-1), which is known to be important for nuclear export of mRNA and proteins. Treatment with a TAP-specific siRNA strikingly shifted cytoplasmic HBc to nucleus, and led to a near 7-fold reduction of viral replication, and a near 10-fold reduction in HBsAg secretion. v) HBc of mutant ARD-II+IV was accumulated predominantly in the nucleus in a mouse model by hydrodynamic delivery. In addition to the revised map of NLS, our results suggest that HBc could shuttle rapidly between nucleus and cytoplasm via a novel TAP-dependent NES.

<A<em>b</em>stractText>The DiRECT trial assessed remission of type 2 dia<em>b</em>etes during a primary care-led weight-management programme. At 1 year, 68 (46%) of 149 intervention participants were in remission and 36 (24%) had achieved at least 15 kg weight loss. The aim of this 2-year analysis is to assess the dura<em>b</em>ility of the intervention effect.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>DiRECT is an open-la<em>b</em>el, cluster-randomised, controlled trial done at primary care practices in the UK. Practices were randomly assigned (1:1) via a computer-generated list to provide an integrated structured weight-management programme (intervention) or <em>b</em>est-practice care in accordance with guidelines (control), with stratification for study site (Tyneside or Scotland) and practice list size (>5700 or ≤5700 people). Allocation was concealed from the study statisticians; participants, carers, and study research assistants were aware of allocation. We recruited individuals aged 20-65 years, with less than 6 years' duration of type 2 dia<em>b</em>etes, BMI 27-45 kg/m², and not receiving insulin <em>b</em>etween July 25, 2014, and Aug 5, 2016. The intervention consisted of withdrawal of antidia<em>b</em>etes and antihypertensive drugs, total diet replacement (825-853 kcal per day formula diet for 12-20 weeks), stepped food reintroduction (2-8 weeks), and then structured support for weight-loss maintenance. The coprimary outcomes, analysed hierarchically in the intention-to-treat population at 24 months, were weight loss of at least 15 kg, and remission of dia<em>b</em>etes, defined as H<em>b</em>A<su<em>b</em>)1c</su<em>b</em>) less than 6·5% (48 mmol/mol) after withdrawal of antidia<em>b</em>etes drugs at <em>b</em>aseline (remission was determined independently at 12 and 24 months). The trial is registered with the ISRCTN registry, num<em>b</em>er 03267836, and follow-up is ongoing.</p><p><div>(<em>b</em>)FINDINGS</<em>b</em>)</div>The intention-to-treat population consisted of 149 participants per group. At 24 months, 17 (11%) intervention participants and three (2%) control participants had weight loss of at least 15 kg (adjusted odds ratio [aOR] 7·49, 95% CI 2·05 to 27·32; p=0·0023) and 53 (36%) intervention participants and five (3%) control participants had remission of dia<em>b</em>etes (aOR 25·82, 8·25 to 80·84; p&<em>lt</em>;0·0001). The adjusted mean difference <em>b</em>etween the control and intervention groups in change in <em>b</em>odyweight was -5·4 kg (95% CI -6·9 to -4·0; p&<em>lt</em>;0·0001) and in H<em>b</em>A<su<em>b</em>)1c</su<em>b</em>) was -4·8 mmol/mol (-8·3 to -1·4 [-0·44% (-0·76 to -0·13)]; p=0·0063), despite only 51 (40%) of 129 patients in the intervention group using anti-dia<em>b</em>etes medication compared with 120 (84%) of 143 in the control group. In a post-hoc analysis of the whole study population, of those participants who maintained at least 10 kg weight loss (45 of 272 with data), 29 (64%) achieved remission; 36 (24%) of 149 participants in the intervention group maintained at least 10 kg weight loss. Serious adverse events were similar to those reported at 12 months, <em>b</em>ut were fewer in the intervention group than in the control group in the second year of the study (nine vs 22).</p><A<em>b</em>stractText>The DiRECT programme sustained remissions at 24 months for more than a third of people with type 2 dia<em>b</em>etes. Sustained remission was linked to the extent of sustained weight loss.</A<em>b</em>stractText><A<em>b</em>stractText>Dia<em>b</em>etes UK.</A<em>b</em>stractText>

The receptors for cholera toxin and Escherichia coli heat-labile toxin (LT) in rabbit small intestinal epithelium were characterized and compared. (i) In vivo studies in ligated intestinal loops showed that whereas LT B subunits could block the fluid secretogenic action of purified LT as well as cholera toxin, cholera toxin B subunits did not inhibit the LT response even when tested in a concentration 100-fold higher than one which gave complete blocking of cholera toxin action. (ii) In vitro studies indicated that isolated intestinal epithelial cells or brush-border membranes could bind about 10-fold more of E. coli LT than of cholera toxin. (iii) All binding sites for cholera toxin in duodenal, jejunal, or ileal mucosal cells or brush-border membranes were extracted by chloroform-methanol-water (4:8:3), which removed lipids quantitatively but did not extract glycoproteins. The extracted cholera toxin binding sites were to greater than 95% recovered in a monosialoganglioside fraction; quantitatively these sites closely corresponded to the concentration of chromatographically identified mucosal GM1 ganglioside (1 nmol of cholera toxin was bound per 1 to 2 nmol of GM1). In contrast, a substantial fraction of mucosal binding sites for E. coli LT remained in the delipidized tissue residue, and these sites had properties consistent with a glycoprotein nature. Thus, whereas cholera toxin appeared to bind highly selectively to GM1 ganglioside receptor sites of rabbit small intestine, E. coli LT bound both to GM1 ganglioside and to a main glycoprotein receptor for which cholera toxin lacks affinity.

Enterotoxigenic Escherichia coli (ETEC) is the most common bacterial cause of diarrhoea in the world, annually affecting up to 400,000,000 children under 5 years of age living in developing countries (DCs). Although ETEC possesses numerous antigens, the relatively conserved colonization factor (CF) antigens and the heat labile enterotoxin (LT) have been associated with protection and most vaccine candidates have exploited these antigens. A safe and effective vaccine against ETEC is a feasible goal as supported by the acquisition of protective immunity. The success of an ETEC vaccine targeting infants and children in DCs will depend on a combination of maximally antigenic vaccine preparations and regimens for their delivery which will produce optimal immune responses to these antigens. Vaccine candidates having a high priority for accelerated development and clinical testing for eventual use in infants would include inactivated ETEC or Shigella hybrids expressing ETEC antigens as well as attenuated ETEC strains which express the major CF antigens and LT toxin B-subunit, as well as attenuated Shigella, Vibrio cholerae and Salmonella typhi hybrids engineered to deliver antigens of ETEC. Candidates for an ETEC vaccine would have to meet the minimal requirement of providing at least 50% protection against severe disease in DCs during the first 2 years of life. The critical roadblock to achieving this goal has not been the science as much as the lack of a sufficiently funded and focused effort to bring it to realization. However, a Product Development Partnership to overcome this hurdle could accelerate the time lines towards when control of ETEC disease in DCs is substantially closer.

Surface lymphotoxin (LT) is a heteromeric complex of LT-alpha and LT-beta chains that binds to the LT-beta receptor (LT-beta-R), a member of the tumor necrosis factor (TNF) family of receptors. The biological function of this receptor-ligand system is poorly characterized. Since signaling through other members of this receptor family can induce cell death, e.g., the TNF and Fas receptors, it is important to determine if similar signaling events can be communicated via the LT-beta-R. A soluble form of the surface complex was produced by coexpression of LT-alpha and a converted form of LT-beta wherein the normally type II LT-beta membrane protein was changed to a type I secreted form. Recombinant LT-alpha 1/beta 2 was cytotoxic to the human adenocarcinoma cell lines HT-29, WiDr, MDA-MB-468, and HT-3 when added with the synergizing agent interferon (IFN) gamma. When immobilized on a plastic surface, anti-LT-beta-R monoclonal antibodies (mAbs) induced the death of these cells, demonstrating direct signaling via the LT-beta-R. Anti-LT-beta-R mAbs were also identified that inhibited ligand-induced cell death, whereas others were found to potentiate the activity of the ligand when added in solution. The human WiDr adenocarcinoma line forms solid tumors in immunocompromised mice, and treatment with an anti-LT-beta-R antibody combined with human IFN-gamma arrested tumor growth. The delineation of a biological signaling event mediated by the LT-beta-R opens a window for further studies on its immunological role, and furthermore, activation of the LT-beta-R may have an application in tumor therapy.

The first three-dimensional chromium(III) dicarboxylate, MIL-53as or Cr(III)(OH) x [O(2)C-C(6)H(4)-CO(2)].[HO(2)C-C(6)H(4)-CO(2)H](0.75), has been obtained under hydrothermal conditions (as: as-synthesized). The free acid can be removed by calcination giving the resulting solid, MIL-53ht or Cr(III)(OH) x [O(2)C-C(6)H(4)-CO(2)]. At room temperature, MIL-53ht adsorbs atmospheric water immediately to give Cr(III)(OH) x [O(2)C-C(6)H(4)-CO(2)] x H(2)O or MIL-53lt (lt: low-temperature form, ht: high-temperature form). Both structures, which have been determined by using X-ray powder diffraction data, are built up from chains of chromium(III) octahedra linked through terephthalate dianions. This creates a three-dimensional structure with an array of one-dimensional large pore channels filled with free disordered terephthalic molecules (MIL-53as) or water molecules (MIL-53lt); when the free molecules are removed, this leads to a nanoporous solid (MIL-53ht) with a Langmuir surface area over 1500 m(2)/g. The transition between the hydrated form (MIL-53lt) and the anhydrous solid (MIL-53ht) is fully reversible and followed by a very high breathing effect (more than 5 A), the pores being clipped in the presence of water molecules (MIL-53lt) and reopened when the channels are empty (MIL-53ht). The thermal behavior of the two solids has been investigated using TGA and X-ray thermodiffractometry. The sorption properties of MIL-53lt have also been studied using several organic solvents. Finally, magnetism measurements performed on MIL-53as and MIL-53lt revealed that these two phases are antiferromagnetic with Néel temperatures T(N) of 65 and 55 K, respectively. Crystal data for MIL-53as is as follows: orthorhombic space group Pnam with a = 17.340(1) A, b = 12.178(1) A, c = 6.822(1) A, and Z = 4. Crystal data for MIL-53ht is as follows: orthorhombic space group Imcm with a = 16.733(1) A, b = 13.038(1) A, c = 6.812(1) A, and Z = 4. Crystal data for MIL-53lt is as follows: monoclinic space group C2/c with a = 19.685(4) A, b = 7.849(1) A, c = 6.782(1) A, beta = 104.90(1) degrees, and Z = 4.

The pathogenesis of aspirin-induced asthma (AIA) has not yet been clearly elucidated, although eicosanoid metabolites appear to play an important role. We hypothesized that levels of eicosanoids in exhaled air condensate are abnormal in patients with AIA and that they change in patients receiving steroid therapy. We measured cysteinyl-leukotrienes (cys-LTs), prostaglandin E(2) (PGE(2)), and leukotriene B(4) (LTB(4)), and also 8-isoprostane as a marker of oxidative stress, by enzyme immunoassay in exhaled breath condensate from patients with AIA (17 steroid naive; mean age, 41 +/- 23 years; FEV(1), 63%pred), 26 patients with aspirin-tolerant asthma (ATA) (11 steroid naive; mean age, 47 +/- 18 years; FEV(1), 69%pred), and 16 healthy subjects (mean age, 45 +/- 17 years; FEV(1), 93%pred). Cys-LTs were significantly higher in steroid-naive patients with AIA compared with steroid-naive patients with ATA and healthy subjects (152.3 +/- 30.4 and 36.6 +/- 7.1 versus 19.4 +/- 2.8 pg/ml; p < 0.05 and p < 0.05, respectively). Steroid-naive patients with AIA also had higher levels of 8-isoprostane than normal subjects (131.8 +/- 31.0 versus 21.9 +/- 4.5 pg/ml; p < 0.05). There were significantly lower levels of both cys-LTs and 8-isoprostanes in steroid-treated patients with AIA. There was no difference in either the PGE(2) or LTB(4) level between the patient groups. This is the first study to show that cys-LTs and 8-isoprostanes are elevated in expired breath condensate of steroid-naive patients with AIA, and that cys-LTs are decreased in steroid-treated patients. Exhaled PGE(2) levels are not reduced, so that it is unlikely that a deficiency of PGE(2) is an important mechanism, whereas exhaled LTB(4) levels are unchanged, indicating an abnormality beyond 5-lipoxygenase.

Mast cells are tissue-resident cells with important functions in allergy and inflammation. Pluripotential hematopoietic stem cells in the bone marrow give rise to committed mast cell progenitors that transit via the blood to tissues throughout the body, where they mature. Knowledge is limited about the factors that release mast cell progenitors from the bone marrow or recruit them to remote tissues. Mouse femoral bone marrow cells were cultured with IL-3 for 2 wk and a range of chemotactic agents were tested on the c-kit(+) population. Cells were remarkably refractory and no chemotaxis was induced by any chemokines tested. However, supernatants from activated mature mast cells induced pronounced chemotaxis, with the active principle identified as leukotriene (LT) B(4). Other activation products were inactive. LTB(4) was highly chemotactic for 2-wk-old cells, but not mature cells, correlating with a loss of mRNA for the LTB(4) receptor, BLTLTB(4). Furthermore, LTB(4) was highly potent in attracting mast cell progenitors from freshly isolated bone marrow cell suspensions. Finally, LTB(4) was a potent chemoattractant for human cord blood-derived immature, but not mature, mast cells. These results suggest an autocrine role for LTB(4) in regulating tissue mast cell numbers.

Prion pathogenesis following oral exposure is thought to involve gut-associated lymphatic tissue, which includes Peyer's patches (PPs) and M cells. Recruitment of activated B lymphocytes to PPs requires alpha(4)beta(7) integrin; PPs of beta 7(-/-) mice are normal in number but are atrophic and almost entirely devoid of B cells. Here we report that minimal infectious dose and disease incubation after oral exposure to logarithmic dilutions of prion inoculum were similar in beta 7(-/-) and wild-type mice, and PPs of both beta 7(-/-) and wild-type mice contained 3-4 log LD(50)/g prion infectivity>> or =125 days after challenge. Despite marked reduction of B cells, M cells were present in beta 7(-/-) mice. In contrast, mice deficient in both tumor necrosis factor and lymphotoxin-alpha (TNF alpha(-/-) x LT alpha(-/-)) or in lymphocytes (RAG-1(-/-), mu MT), in which numbers of PPs are reduced in number, were highly resistant to oral challenge, and their intestines were virtually devoid of prion infectivity at all times after challenge. Therefore, lymphoreticular requirements for enteric and for intraperitoneal uptake of prions differ from each other. Although susceptibility to prion infection following oral challenge correlates with the number of PPs, it is remarkably independent of the number of PP-associated lymphocytes.

<A<em>b</em>stractText>The selective TRK inhi<em>b</em>itor larotrectini<em>b</em> was approved for paediatric and adu<em>lt</em> patients with advanced TRK fusion-positive solid tumours <em>b</em>ased on a primary analysis set of 55 patients. The aim of our analysis was to explore the efficacy and long-term safety of larotrectini<em>b</em> in a larger population of patients with TRK fusion-positive solid tumours.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>Patients were enrolled and treated in a phase 1 adu<em>lt</em>, a phase 1/2 paediatric, or a phase 2 adolescent and adu<em>lt</em> trial. Some eligi<em>b</em>ility criteria differed <em>b</em>etween these studies. For this pooled analysis, eligi<em>b</em>le patients were aged 1 month or older, with a locally advanced or metastatic non-CNS primary, TRK fusion-positive solid tumour, who had received standard therapy previously if availa<em>b</em>le. This analysis set includes the 55 patients on which approval of larotrectini<em>b</em> was <em>b</em>ased. Larotrectini<em>b</em> was administered orally (capsule or liquid formulation), on a continuous 28-day schedule, to adu<em>lt</em>s mostly at a dose of 100 mg twice daily, and to paediatric patients mostly at a dose of 100 mg/m² (maximum of 100 mg) twice daily. The primary endpoint was o<em>b</em>jective response as assessed <em>b</em>y local investigators in an intention-to-treat analysis. Contri<em>b</em>uting trials are registered with ClinicalTrials.gov, NCT02122913 (active not recruiting), NCT02637687 (recruiting), and NCT02576431 (recruiting).</p><A<em>b</em>stractText>Between May 1, 2014, and Fe<em>b</em> 19, 2019, 159 patients with TRK fusion-positive cancer were enrolled and treated with larotrectini<em>b</em>. Ages ranged from less than 1 month to 84 years. The proportion of patients with an o<em>b</em>jective response according to investigator assessment was 121 (79%, 95% CI 72-85) of 153 evalua<em>b</em>le patients, with 24 (16%) having complete responses. In a safety population of 260 patients treated regardless of TRK fusion status, the most common grade 3 or 4 larotrectini<em>b</em>-related adverse events were increased alanine aminotransferase (eight [3%] of 260 patients), anaemia (six, 2%), and decreased neutrophil count (five [2%]). The most common larotrectini<em>b</em>-related serious adverse events were increased alanine aminotransferase (two [&<em>lt</em>;1%] of 260 patients), increased aspartate aminotransferase (two [&<em>lt</em>;1%]), and nausea (two [&<em>lt</em>;1%]). No treatment-related deaths occurred.</A<em>b</em>stractText><A<em>b</em>stractText>These data confirm that TRK fusions define a unique molecular su<em>b</em>group of advanced solid tumours for which larotrectini<em>b</em> is highly active. Safety data indicate that long-term administration of larotrectini<em>b</em> is feasi<em>b</em>le.</A<em>b</em>stractText><A<em>b</em>stractText>Bayer and Loxo Oncology.</A<em>b</em>stractText>

Recent studies revealed that the lymphotoxin/lymphotoxin beta receptor (LT)/LTbetaR system activates the noncanonical nuclear factor-kappaB (NF-kappaB) signaling pathway involving I kappa B kinase 1/I kappa B kinase alpha (IKK1/IKKalpha) and NF-kappaB-inducing kinase (NIK) to direct processing of the nfkappab2 protein p100 to yield RelB:p52 complexes. Despite the biochemical evidence, LT-, RelB-, p52-deficient mice show discrepant phenotypes. We now demonstrate that p105/p50 also constitutes an important pathway for LTbetaR signaling. Our studies revealed that mice deficient in either p50 or p52 have defects in the formation of inguinal lymph nodes (LNs), but that the complete defect in lymph node formation and splenic microarchitecture seen in LT-deficient mice is recapitulated only in mice deficient in both p50 and p52. Biochemically, we find not only that both p50- and p52-containing NF-kappaB activities are induced by LTbetaR signaling, but that the induction of NF-kappaB-containing complexes by LTbetaR engagement is perturbed in single knockouts. Importantly, the LTbetaR can additionally activate the less well-characterized p52:RelA and p50:RelB pathways, which play pivotal roles in vivo for the development and organization of lymphoid structures. Our genetic, cellular, and molecular evidence points toward a model of LT-mediated NF-kappaB regulation in which p105/p50 and p100/p52 have distinct and coordinating molecular specificities but differ in the upstream signaling pathways that regulate them.

BACKGROUND

Colonoscopy preparation regimens are often poorly tolerated because they require use of large-volume bowel preparation solution and diet restrictions for adequate cleansing. This study evaluated the efficacy and tolerability of a split-dose polyethylene glycol-electrolyte solution plus bisacodyl and a regular diet.

METHODS

A total of 187 patients (104 men, 83 women; age range 18-91 years) were randomly assigned to receive either 3 L of polyethylene glycol-electrolyte solution (n = 96; Group A) with a liquid diet on the day before colonoscopy, or 2 L of polyethylene glycol-electrolyte solution, one tablet of bisacodyl, and a minimally restricted diet on the day before colonoscopy, and then 1 L of the same solution on the day of colonoscopy (n = 91; Group B). Acceptability, adverse events, and willingness to retake the preparation were assessed by questionnaire. The quality of the preparation was graded by an endoscopist, blinded to the type of preparation, by using a previously described scale (excellent to poor).

RESULTS

There were 96 patients in Group A and 91 in Group B. Colon cleansing was significantly better in Group B with regard to the overall quality of the preparation (p lt; 0.05). Compliance was significantly higher in Group B as evidenced by the lower number of patients who discontinued the preparation (4 vs. 15; p = 0.02) because of side effects such as nausea or vomiting. The degree of discomfort, adverse events, and willingness to retake the preparation were not significantly different between the groups.

CONCLUSIONS

Colonic preparation with split-dose polyethylene glycol-electrolyte provided better quality colon cleansing and higher compliance, with less dietary restrictions, than preparation with whole-dose polyethylene glycol-electrolyte.