Mitigation of acetaminophen-induced liver toxicity by the novel phosphatidylinositol 3-kinase inhibitor alpelisib

The sterile inflammatory response mediated by Toll-like receptors (TLRs) 4 and 9 is implicated in the massive hepatic damage caused by acetaminophen (APAP)-overdose. There is a crosstalk between TLR-dependent signaling with other intracellular kinases like phosphatidylinositol 3-kinases (PI3Ks). Nevertheless, the detailed role of PI3Kα is still unknown in hepatic sterile inflammation. Accordingly, the effect of the novel PI3Kα inhibitor alpelisib was investigated in the setting of APAP-driven sterile inflammation in the liver. This was examined by pretreating mice with alpelisib (5 and 10 mg/kg, oral) 2 h before APAP (500 mg/kg, i.p.)-intoxication. The results indicated that alpelisib dose-dependently lowered APAP-induced escalation in serum liver function biomarkers and hepatic necroinflammation score. Alpelisib also attenuated APAP-induced rise in cleaved caspase 3 and proliferating cell nuclear antigen (PCNA) in the liver hepatocytes, as indices for apoptosis and proliferation. Mechanistically, inhibition of PI3Kα by alpelisib limited APAP-induced overproduction of the pro-inflammatory tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in the blood circulation via switching off the activation of several signal transduction proteins, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), signal transducer and activator of transcription-3 (Stat-3), glycogen Synthase Kinase (GSK)-3β and nuclear factor (NF)-κB. Alpelisib also impaired APAP-instigated immune cell infiltration in the liver via reducing systemic granulocyte/macrophage-colony stimulating factor (GM-CSF) release and reversed APAP-induced abnormalities in the systemic and hepatic levels of the anti-inflammatory IL-10 and IL-22. In conclusion, selective modulation of the PI3Kα activity by alpelisib can hinder the inflammatory response and infiltration of immune cells occurring by APAP-hepatotoxicity

Bioactive brominated oxindole alkaloids from the Red Sea sponge Callyspongia siphonella

In the present study, LC-HRESIMS-assisted dereplication along with bioactivity-guided isolation led to targeting two brominated oxindole alkaloids (compounds 1 and 2) which probably play a key role in the previously reported antibacterial, antibiofilm, and cytotoxicity of Callyspongia siphonella crude extracts. Both metabolites showed potent antibacterial activity against Gram-positive bacteria, Staphylococcus aureus (minimum inhibitory concentration (MIC) = 8 and 4 µg/mL) and Bacillus subtilis (MIC = 16 and 4 µg/mL), respectively. Furthermore, they displayed moderate biofilm inhibitory activity in Pseudomonas aeruginosa (49.32% and 41.76% inhibition, respectively), and moderate in vitro antitrypanosomal activity (13.47 and 10.27 µM, respectively). In addition, they revealed a strong cytotoxic effect toward different human cancer cell lines, supposedly through induction of necrosis. This study sheds light on the possible role of these metabolites (compounds 1 and 2) in keeping fouling organisms away from the sponge outer surface, and the possible applications of these defensive molecules in the development of new anti-infective agents

Laparoscopy in management of appendicitis in high-, middle-, and low-income countries: a multicenter, prospective, cohort study.

BACKGROUND: Appendicitis is the most common abdominal surgical emergency worldwide. Differences between high- and low-income settings in the availability of laparoscopic appendectomy, alternative management choices, and outcomes are poorly described. The aim was to identify variation in surgical management and outcomes of appendicitis within low-, middle-, and high-Human Development Index (HDI) countries worldwide. METHODS: This is a multicenter, international prospective cohort study. Consecutive sampling of patients undergoing emergency appendectomy over 6 months was conducted. Follow-up lasted 30 days. RESULTS: 4546 patients from 52 countries underwent appendectomy (2499 high-, 1540 middle-, and 507 low-HDI groups). Surgical site infection (SSI) rates were higher in low-HDI (OR 2.57, 95% CI 1.33-4.99, p = 0.005) but not middle-HDI countries (OR 1.38, 95% CI 0.76-2.52, p = 0.291), compared with high-HDI countries after adjustment. A laparoscopic approach was common in high-HDI countries (1693/2499, 67.7%), but infrequent in low-HDI (41/507, 8.1%) and middle-HDI (132/1540, 8.6%) groups. After accounting for case-mix, laparoscopy was still associated with fewer overall complications (OR 0.55, 95% CI 0.42-0.71, p < 0.001) and SSIs (OR 0.22, 95% CI 0.14-0.33, p < 0.001). In propensity-score matched groups within low-/middle-HDI countries, laparoscopy was still associated with fewer overall complications (OR 0.23 95% CI 0.11-0.44) and SSI (OR 0.21 95% CI 0.09-0.45). CONCLUSION: A laparoscopic approach is associated with better outcomes and availability appears to differ by country HDI. Despite the profound clinical, operational, and financial barriers to its widespread introduction, laparoscopy could significantly improve outcomes for patients in low-resource environments. TRIAL REGISTRATION: NCT02179112

Global economic burden of unmet surgical need for appendicitis

Background: There is a substantial gap in provision of adequate surgical care in many low-and middle-income countries. This study aimed to identify the economic burden of unmet surgical need for the common condition of appendicitis. Methods: Data on the incidence of appendicitis from 170 countries and two different approaches were used to estimate numbers of patients who do not receive surgery: as a fixed proportion of the total unmet surgical need per country (approach 1); and based on country income status (approach 2). Indirect costs with current levels of access and local quality, and those if quality were at the standards of high-income countries, were estimated. A human capital approach was applied, focusing on the economic burden resulting from premature death and absenteeism. Results: Excess mortality was 4185 per 100 000 cases of appendicitis using approach 1 and 3448 per 100 000 using approach 2. The economic burden of continuing current levels of access and local quality was US $92 492 million using approach 1 and$73 141 million using approach 2. The economic burden of not providing surgical care to the standards of high-income countries was $95 004 million using approach 1 and$75 666 million using approach 2. The largest share of these costs resulted from premature death (97.7 per cent) and lack of access (97.0 per cent) in contrast to lack of quality. Conclusion: For a comparatively non-complex emergency condition such as appendicitis, increasing access to care should be prioritized. Although improving quality of care should not be neglected, increasing provision of care at current standards could reduce societal costs substantially

Pooled analysis of WHO Surgical Safety Checklist use and mortality after emergency laparotomy

Background The World Health Organization (WHO) Surgical Safety Checklist has fostered safe practice for 10 years, yet its place in emergency surgery has not been assessed on a global scale. The aim of this study was to evaluate reported checklist use in emergency settings and examine the relationship with perioperative mortality in patients who had emergency laparotomy. Methods In two multinational cohort studies, adults undergoing emergency laparotomy were compared with those having elective gastrointestinal surgery. Relationships between reported checklist use and mortality were determined using multivariable logistic regression and bootstrapped simulation. Results Of 12 296 patients included from 76 countries, 4843 underwent emergency laparotomy. After adjusting for patient and disease factors, checklist use before emergency laparotomy was more common in countries with a high Human Development Index (HDI) (2455 of 2741, 89.6 per cent) compared with that in countries with a middle (753 of 1242, 60.6 per cent; odds ratio (OR) 0.17, 95 per cent c.i. 0.14 to 0.21, P <0001) or low (363 of 860, 422 per cent; OR 008, 007 to 010, P <0.001) HDI. Checklist use was less common in elective surgery than for emergency laparotomy in high-HDI countries (risk difference -94 (95 per cent c.i. -11.9 to -6.9) per cent; P <0001), but the relationship was reversed in low-HDI countries (+121 (+7.0 to +173) per cent; P <0001). In multivariable models, checklist use was associated with a lower 30-day perioperative mortality (OR 0.60, 0.50 to 073; P <0.001). The greatest absolute benefit was seen for emergency surgery in low- and middle-HDI countries. Conclusion Checklist use in emergency laparotomy was associated with a significantly lower perioperative mortality rate. Checklist use in low-HDI countries was half that in high-HDI countries.Peer reviewe

Mortality of emergency abdominal surgery in high-, middle- and low-income countries

Background: Surgical mortality data are collected routinely in high-income countries, yet virtually no low- or middle-income countries have outcome surveillance in place. The aim was prospectively to collect worldwide mortality data following emergency abdominal surgery, comparing findings across countries with a low, middle or high Human Development Index (HDI). Methods: This was a prospective, multicentre, cohort study. Self-selected hospitals performing emergency surgery submitted prespecified data for consecutive patients from at least one 2-week interval during July to December 2014. Postoperative mortality was analysed by hierarchical multivariable logistic regression. Results: Data were obtained for 10 745 patients from 357 centres in 58 countries; 6538 were from high-, 2889 from middle- and 1318 from low-HDI settings. The overall mortality rate was 1⋅6 per cent at 24 h (high 1⋅1 per cent, middle 1⋅9 per cent, low 3⋅4 per cent; P < 0⋅001), increasing to 5⋅4 per cent by 30 days (high 4⋅5 per cent, middle 6⋅0 per cent, low 8⋅6 per cent; P < 0⋅001). Of the 578 patients who died, 404 (69⋅9 per cent) did so between 24 h and 30 days following surgery (high 74⋅2 per cent, middle 68⋅8 per cent, low 60⋅5 per cent). After adjustment, 30-day mortality remained higher in middle-income (odds ratio (OR) 2⋅78, 95 per cent c.i. 1⋅84 to 4⋅20) and low-income (OR 2⋅97, 1⋅84 to 4⋅81) countries. Surgical safety checklist use was less frequent in low- and middle-income countries, but when used was associated with reduced mortality at 30 days. Conclusion: Mortality is three times higher in low- compared with high-HDI countries even when adjusted for prognostic factors. Patient safety factors may have an important role. Registration number: NCT02179112 (http://www.clinicaltrials.gov)

Global variation in anastomosis and end colostomy formation following left-sided colorectal resection

Background End colostomy rates following colorectal resection vary across institutions in high-income settings, being influenced by patient, disease, surgeon and system factors. This study aimed to assess global variation in end colostomy rates after left-sided colorectal resection. Methods This study comprised an analysis of GlobalSurg-1 and -2 international, prospective, observational cohort studies (2014, 2016), including consecutive adult patients undergoing elective or emergency left-sided colorectal resection within discrete 2-week windows. Countries were grouped into high-, middle- and low-income tertiles according to the United Nations Human Development Index (HDI). Factors associated with colostomy formation versus primary anastomosis were explored using a multilevel, multivariable logistic regression model. Results In total, 1635 patients from 242 hospitals in 57 countries undergoing left-sided colorectal resection were included: 113 (6·9 per cent) from low-HDI, 254 (15·5 per cent) from middle-HDI and 1268 (77·6 per cent) from high-HDI countries. There was a higher proportion of patients with perforated disease (57·5, 40·9 and 35·4 per cent; P < 0·001) and subsequent use of end colostomy (52·2, 24·8 and 18·9 per cent; P < 0·001) in low- compared with middle- and high-HDI settings. The association with colostomy use in low-HDI settings persisted (odds ratio (OR) 3·20, 95 per cent c.i. 1·35 to 7·57; P = 0·008) after risk adjustment for malignant disease (OR 2·34, 1·65 to 3·32; P < 0·001), emergency surgery (OR 4·08, 2·73 to 6·10; P < 0·001), time to operation at least 48 h (OR 1·99, 1·28 to 3·09; P = 0·002) and disease perforation (OR 4·00, 2·81 to 5·69; P < 0·001). Conclusion Global differences existed in the proportion of patients receiving end stomas after left-sided colorectal resection based on income, which went beyond case mix alone

Entwicklung CD40 gerichteter bifunktioneller scFv-TRAIL Fusionsproteine die TRAILR1- und TRAILR2-spezifischen Zelltod und dendritischen Zellaktivierung induzieren

TRAIL is a member of TNF superfamily and mediates apoptosis by binding to two DRs, TRAILR1 and TRAILR2. Despite the fact that there are other TRAILRs, TRAILR1 and TRAILR2 receive the major research interest due to their ability to trigger apoptosis and their possible use as targets in tumor therapy. Due to the potential advantages of TRAILR1- or TRAILR2-specific targeting, we investigated recently published TRAIL DR-specific mutants, one conferring specificity for TRAILR1 (TRAILmutR1) and one for TRAILR2 (TRAILmutR2). It was well proved in this work that TRAILmutR1 shows specific binding to TRAILR1 and no specific binding to TRAILR2. TRAILmutR2 vice versa shows specific binding to TRAILR2 and no significant binding to TRAILR1. Moreover, these mutants were able to induce caspase activation and cell death in a TRAILR1/2-specific manner. Moreover, the enhancement of TRAILR2-induced apoptosis by secondary oligomerization of soluble wild-type TRAIL was confirmed for the TRAILR2-specifc TRAIL mutant and similar findings were made with the TRAILR1-specific TRAIL mutant. The soluble form of TRAIL exhibits weak apoptotic activity as compared to transmembrane TRAIL. Therefore, there is the challenge in clinical research to improve the activity of soluble TRAIL. A second strategy besides the above mentioned oligomerization to improve soluble TRAIL activity is anchoring of the molecule to the cell surface, e.g. through the genetic fusion with a scFv domain recognizing a cell surface antigen. In this work, we generated fusion proteins of TRAIL, TRAILmutR1 and TRAILmutR2 with a scFv recognizing CD40 (scFv:G28). Initially, we analyzed the functionality of both the TRAIL domain and the scFv:G28 domain of the corresponding fusion proteins. TRAIL functionality was well proved through its ability to induce cell death in TRAIL sensitive cells such as Jurkat cells, provided that scFv:G28-TRAIL fusion proteins were oligomerized by anti-Flag mAb M2. Concerning the scFv:G28 domain, the fusion proteins showed enhanced binding affinity to cell lines expressing CD40 as compared to their parental CD40-negative cells. Consistent with previous studies investigating TRAIL fusion proteins with other cell surface antigen-targeting scFvs, the scFv:G28 fusion proteins with TRAIL, TRAILmutR1 and TRAILmutR2 showed enhanced induction of cell death in a CD40-dependent manner. Moreover, our results revealed that these fusion proteins have a significant paracrine apoptotic effect on CD40-negative bystander cells upon anchoring to CD40-positive cells which are TRAIL resistant. Thus, the current work provides for the first time scFv fusion proteins of TRAIL and TRAILR1- and TRAILR2-specific TRAIL mutants with CD40-restricted activity. These fusion proteins provide the advantage of attenuating the off-target effects and the potential side effects of per se highly active TRAIL variants on one hand due to the CD40-binding dependent enhancement of activity and on the other hand due to the differential use of TRAILR1 and TRAILR2. CD40 represents a tumor associated marker which is expressed on many tumor cells but also on immune cells. Therefore, the last part of this work focused on the analysis of the ability of scFv:G28-TRAIL fusion proteins to induce CD40 signaling both in tumor cells and also in immune cells. It turned out that the scFv:G28-TRAIL fusion proteins are able to induce CD40 signaling in CD40-positive tumor cells but especially also in immune cells such as iDCs leading to their maturation and further activation of immune responses. Taken together, this work provides novel bifunctional scFv-TRAIL fusion proteins which combine the induction of apoptosis via TRAIL DR with stimulation of CD40 signaling which possibly enhances antitumor immunity.TRAIL ist ein Mitglied der TNF-Superfamilie und vermittelt Apoptose durch die Aktivierung der Todesrezeptoren, TRAILR1 und TRAILR2. Obwohl es weitere TRAIL-Rezeptoren gibt, liegt das Hauptaugenmerk auf den beiden Apoptose induzierenden Rezeptoren TRAILR1 und TRAILR2 auf Grund ihrer möglichen Anwendung in der Tumortherapie. Wegen der möglichen Vorteile eines spezifischen TRAILR1- und TRAILR2-Targetings, haben wir kürzlich publizierte TRAIL-Todesrezeptor spezifische TRAIL Mutanten untersucht, von denen eine spezifisch für TRAILR1 (TRAILmutR1) und die andere spezifisch für TRAILR2 (TRAILmutR2) ist. Es konnte in dieser Arbeit sehr gut belegt werden, dass TRAILmutR1 spezifisch an TRAILR1 bindet und keine Bindung an TRAILR2 zeigte. Dem entsprechend zeigte die Variante TRAILmutR2 nur eine spezifische Bindung an TRAILR2 und keine signifikante Bindung an TRAILR1. Des Weiteren waren die Mutanten in der Lage, die Caspase-Aktivierung und den Zelltod TRAILR1/2-abhängig zu induzieren. Außerdem konnte eine Erhöhung der TRAILR2-induzierten Apoptose durch eine sekundäre Oligomerisierung der TRAILR2-spezifische TRAIL-Mutante erzielt werden. Ähnliche Ergebnisse zeigte die TRAILR1-spezifische TRAIL-Mutante. Um die Aktivität des löslichen TRAIL Oligomerisierung unabhängig zu erhöhen, wurden in dieser Arbeit TRAIL-Fusionsproteine mit einem scFv (scFv:G28), der CD40 erkennt generiert. In Übereinstimmung mit früheren Studien, die mit TRAIL-Fusionsproteinen von anderen Zelloberflächenantigen-spezifischen scFvs wurden, zeigten die CD40-spezifischen scFv:G28 Fusionsproteine mit TRAIL, TRAILmutR1 und TRAILmutR2 eine verstärkte CD40-abhängige Induktion des Zelltods. Darüber hinaus zeigten unsere Ergebnisse, dass diese Fusionsproteine nach Bindung an CD40-positive Zellen einen parakrinen apoptotischen Effekt, auf umliegende CD40-negative Zellen haben. Diese Arbeit beschreibt somit zum ersten Mal scFv-TRAIL Fusionsproteine mit einer CD40-abhängigen TRAILR1- und TRAILR2-spezifischen Aktivität. CD40 repräsentiert einen tumorassoziierten Marker, der in vielen Tumorzellen aber auch in Zellen des Immunsystems exprimiert wird. Aus diesem Grund fokussierte sich der zweite Teil dieser Arbeit auf die Analyse der Fähigkeit der scFv:G28-TRAIL Fusionsproteine, CD40-Signaling sowohl in Tumor- als auch in Immunzellen zu stimulieren. Es konnte festgestellt werden, dass die scFv:G28-TRAIL Fusionsproteine in der Lage sind, CD40-Signaling in CD40-positiven Tumorzellen, aber auch in Immunzellen, z.B. in iDCs, in denen die ScFv-TRAIL Fusionsproteine die Reifung und Aktivierung induzieren ohne Zelltod auszulösen. Zusammengefasst beschreibt diese Arbeit neue bifunktionelle scFv-TRAIL Fusionsproteine, die die Induktion der Apoptose via TRAIL-Todesrezeptoren und die Stimulation des kostimulatorischen CD40-Moleküls kombinieren, was zu einer synergistischen dualen Antitumor-Aktivität führen kann

Vitamin D potentiates anti-tumor activity of 5-fluorouracil via modulating caspase-3 and TGF-β1 expression in hepatocellular carcinoma-induced in rats

We investigated the role of vitamin D (Vit D) alone and in combination with 5-fluorouracil (5-FU) in thioacetamide (TAA)-induced hepatocellular carcinoma (HCC) in rats. Fifty male Sprague–Dawley rats were randomized into a control group and 4 groups that received TAA (200 mg/kg, i.p.) twice per week for 16 weeks. These 4 groups were further divided as follows: HCC group; 5-FU group (75 mg/kg, i.p., once weekly for 3 weeks starting from the 12th week); Vit D group (200 IU/kg daily by oral tube for 16 weeks); and 5-FU + Vit D group (received the previously mentioned dosage regimens of 5-FU and Vit D). HCC was detected by histopathological changes in liver sections and the elevation of serum α-fetoprotein (AFP). Treatment with 5-FU + Vit D significantly decreased gene expression of nuclear factor erythroid 2-related factor 2 (NrF2) and transforming growth factor β1 (TGF-β1) at both the gene and protein level and serum AFP concentrations in comparison with their corresponding monotherapy. Moreover, 5-FU + Vit D treatment enhanced apoptosis by increasing caspase-3 gene and protein expression. Conclusively, Vit D enhances antitumor activity of 5-FU in an HCC-induced model and improves liver function of treated animals. Combination therapy in a TAA-induced HCC rat model was more effective than 5-FU or Vit D through the modulation of TGF-β1, caspase-3, and NrF2 expressions.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author