Refsgaardcunningham7144

Biotechnology and nanotechnology are important tools for understanding biochemical pathways. They can be used efficiently for stimulating and increasing the production of secondary metabolites in medicinal plants. The present study aimed to identify the γ-terpinene synthase gene (CcTPS2) as an effective contributor to the biosynthetic pathway of monoterpenes. The effects of silver nanoparticles (AgNPs; 50 and 100 mg l- 1) and time (24 and 48 h) were examined on secondary metabolites in cell suspension cultures of Carum carvi. This involved the identification, isolation, and sequencing of a partial sequence in the CcTPS2 gene of C. carvi. The genomic sequence of CcTPS2 comprised 292 bp which were organized into two exons (110 and 82 bp) and one intron (100 bp), while the cDNA was 192 bp. In the scale of nucleotides, the CcTPS2 gene showed 96% similarity with the TPS2 gene of Oliveria decumbens. We generated sequence data of the CcTPS2 gene for the first time in this species, thereby enabling further developments in understanding the molecular mechanisms responsible for terpene biosynthesis and other chemical derivatives in C. carvi. The results of GC/MS and GC/FID showed that AgNPs strongly affected the secondary metabolites in cell suspension cultures of C. carvi. According to the results, the AgNPs (50 mg l- 1) increased p-cymene and carvone contents in comparison with the control. The exposure of plants to 100 mg l- 1 AgNPs induced the production of thymol and carvacrol. The results of real-time PCR revealed that the exposure of plants to 100 mg l- 1 AgNPs caused a significant upregulation of CcTPS2 expression for 24 h. These cell suspension cultures were elicited by AgNPs, the application of which proved as an effective method to improve the production of secondary metabolites in vitro.Precursor feeding is a potential strategy for increasing specialized metabolite production in plant cell culture systems. In the present study, cell suspension cultures were developed and subsequently evaluated for precursor feeding investigations. Cell suspension cultures were established in Murashige and Skoog (MS) medium containing 0.5 mg/L thidiazuron (TDZ) + 1 mg/L α-naphthalene acetic acid (NAA). The growth biomass and metabolite pattern were analyzed to identify specific culture days required for prolific biomass production. The maximum cell dry weight (DW) was observed in leaf cell suspension (1.22 g/100 mL) and root cell suspension culture (1.12 g/100 mL) on day 21. Afterward, the effect of precursor concentrations (tyrosol; 0.5, 1, 2, and 3 mM) along with two light regimes, photoperiod (16L/8D h, 70 µmol/m2/s) and dark (24 h), was evaluated for cell growth and metabolite accumulation. The results revealed that leaf cell suspension treated with 3 mM tyrosol concentration detected maximum salidroside content (26.05 mg/g DW) on day 15, incubated under photoperiod (16L/8D h) condition. Similarly, under photoperiod (16L/8D h), root cell suspension treated with 3 mM tyrosol produced maximum salidroside content (26.62 mg/g DW) on day 12. Moreover, the total phenolics content increased significantly (44.21 mg/g DW) on day 12 in 3 mM tyrosol treatment under photoperiod (16L/8D h). However, precursor concentrations did not influence the total flavonoids content. The present investigation suggests that the immediate pathway precursor, tyrosol, has a strong effect on enhanced production of salidroside, irrespective of explant type and light regimes.One of the hypothesized mechanisms of sudden cardiac death in humans is an arrhythmia precipitated by increased sympathetic outflow to a compromised heart. The stellate ganglia provide the main sympathetic innervation to the heart, where the left stellate ganglion appears to play a role in arrhythmogenesis. Case reports of sudden cardiac death have described left stellate ganglion inflammation but no larger studies have been performed. Thus, we have specifically assessed whether the left stellate ganglion was inflamed in those dying from sudden cardiac death versus other causes of death. Thirty-one left stellate ganglia were resected from cadavers diagnosed with sudden cardiac deaths and compared with 18 ganglia from cadavers diagnosed with non-sudden cardiac deaths. Ganglia were stained with hematoxylin and eosin and lymphocytic aggregates compared. The proportion of left stellate ganglion inflammation (77%) was significantly higher in deaths from sudden cardiac deaths than non-sudden cardiac deaths (33%). This study provides information on a previously recognized, but understudied, structure that may help understand sudden cardiac death. We found high prevalence of stellate ganglion inflammation and propose that this may trigger sympathetic storms.Autoimmune diseases of the peripheral nervous system have so far been treated mainly with exogenous high-dose intravenous immunoglobulins (IVIg), that act through several mechanisms, including neutralization of pathogenic autoantibodies, modulation of lymphocyte activity, interference with antigen presentation, and interaction with Fc receptors, cytokines, and the complement system. Other therapeutic strategies have recently been developed, in part to address the increasing shortage of IVIg, prime among which is the use of B cell depleting monoclonal antibodies, or small molecule inhibitors targeting the B-cell specific kinases. Rituximab, a chimeric monoclonal antibody against CD20 + B lymphocytes, is currently the most used, especially in anti-MAG antibody neuropathy and autoimmune neuropathies with antibodies to nodal/paranodal antigens that are unresponsive to IVIg. After several reports of its efficacy in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), rituximab is currently under invesy rozanolixizumab is currently being assessed in phase 2 trials in CIDP. However, none of the abovementioned monoclonal antibodies is currently approved for treatment of any immune-mediated neuropathies. While more specific and individualized therapies are being developed, the possibility of combined treatments targeting different pathogenic mechanisms deserves consideration as well.Ponatinib is effective in adults with Philadelphia chromosome-positive (Ph+) leukemia, resistant or intolerant to second-generation tyrosine kinase inhibitors, but there are limited data on its use in children. The clinical courses of nine pediatric patients with Ph+ acute lymphoblastic leukemia (Ph+ ALL) and four with chronic myeloid leukemia (CML) who received ponatinib therapy were retrospectively reviewed. The median age at the start of ponatinib therapy was 12 years (range 8-16 years). Nine patients were male and four were female. Six patients received ponatinib alone, three received ponatinib with prednisolone, one received ponatinib with rituximab intrathecal therapy, and one received ponatinib with conventional chemotherapy. Two patients received ponatinib both alone and in combination with chemotherapy. The median dose and duration of ponatinib were 16.9 mg/m2 (7-34.3) and 1.1 months (0.2-22.7), respectively. Six patients with Ph+ ALL and two with CML responded to ponatinib. One of the eight patients who received ponatinib alone had grade 4 increased lipase levels. Grade 3 non-hematologic toxicities included elevated alanine aminotransferase levels (25%), elevated aspartate aminotransferase levels (25%), elevated gamma-glutamyl transferase levels (12.5%), hypertension (12.5%), and polymorphic erythema (12.5%). Ponatinib may be safe and effective in pediatric patients with Ph+ leukemia.

Severe alcoholic hepatitis (SAH) presenting as acute-on-chronic liver failure (ACLF) carries a high short-term mortality. Alteration of gut microbiota is a crucial component implicated in its pathogenesis, whose modulation has been suggested as a potential therapeutic tool. We evaluated the safety of fecal microbiota transplantation (FMT) and its efficacy in improving short-term survival and clinical severity scores in patients with SAH-ACLF.

Thirty-three patients [13 in the FMT arm; 20 in the standard of care arm (SOC)] with SAH-ACLF were included in this open-label study. A single FMT session was administered as a freshly prepared stool suspension from pre-identified healthy family member stool donors through a nasojejunal tube. Patients were followed up on days 7, 28, and 90.

Survival at 28 and 90 days was significantly better in the FMT arm (100% versus 60%, p = 0.01; 53.84% versus 25%, p = 0.02). Hepatic encephalopathy resolved in 100% versus 57.14% (FMT versus SOC, p = 0.11) patients, while ascites resolved in 100% versus 40% survivors (p = 0.04). Major adverse event rates, including spontaneous bacterial peritonitis and gastrointestinal bleeding, were similar in both groups (p = 0.77; p = 0.70). Infigratinib mouse Median IL1beta decreased by21.39% (IQR -73.67 to 7.63) in the FMT group, whereas it increased in the SOC by 27.44% (IQR -0.88 to 128.11) (p = 0.01). Percentage changes in bilirubin and ALT between baseline and day 7 emerged as predictors of 90-day mortality.

FMT is safe, improves short-term and medium-term survival, and leads to improvement in clinical severity scores in patients with SAH-ACLF.

NCT03827772 available from http//clinicaltrials.gov/ct2/show/NCT03827772 CTRI Reference number CTRI/2019/02/017538 dated 7 February 2019.

NCT03827772 available from http//clinicaltrials.gov/ct2/show/NCT03827772 CTRI Reference number CTRI/2019/02/017538 dated 7 February 2019.

Microvascular invasion (MVI) is essential for the management of hepatocellular carcinoma (HCC). However, MVI is hard to evaluate in patients without sufficient peri-tumoral tissue samples, which account for over a half of HCC patients.

We established an MVI deep-learning (MVI-DL) model with a weakly supervised multiple-instance learning framework, to evaluate MVI status using only tumor tissues from the histological whole slide images (WSIs). A total of 350 HCC patients (2917 WSIs) from the First Affiliated Hospital of Sun Yat-sen University (FAHSYSU cohort) were divided into a training and test set. One hundred and twenty patients (504 WSIs) from Dongguan People's Hospital and Shunde Hospital of Southern Medical University (DG-SD cohort) formed an external test set. Unsupervised clustering and class activation mapping were applied to visualize the key histological features.

In the FAHSYSU and DG-SD test set, the MVI-DL model achieved an AUC of 0.904 (95% CI 0.888-0.920) and 0.871 (95% CI 0.837-0.905), respectively. Visualization results showed that macrotrabecular architecture with rich blood sinus, rich tumor stroma and high intratumor heterogeneity were identified as the key features associated with MVI ( +), whereas severe immune infiltration and highly differentiated tumor cells were associated with MVI (-). In the simulation of patients with only one WSI or biopsies only, the AUC of the MVI-DL model reached 0.875 (95% CI 0.855-0.895) and 0.879 (95% CI 0.853-0.906), respectively.

The effective, interpretable MVI-DL model has potential as an important tool with practical clinical applicability in evaluating MVI status from the tumor areas on the histological slides.

The effective, interpretable MVI-DL model has potential as an important tool with practical clinical applicability in evaluating MVI status from the tumor areas on the histological slides.