SHP-2 and PD-1-SHP-2 signaling regulate myeloid cell differentiation and antitumor responses.

The inhibitory receptor PD-1 suppresses T cell activation by recruiting the phosphatase SHP-2. However, mice with a T-cell-specific deletion of SHP-2 do not have improved antitumor immunity. Here we showed that mice with conditional targeting of SHP-2 in myeloid cells, but not in T cells, had diminished tumor growth. RNA sequencing (RNA-seq) followed by gene set enrichment analysis indicated the presence of polymorphonuclear myeloid-derived suppressor cells and tumor-associated macrophages (TAMs) with enriched gene expression profiles of enhanced differentiation, activation and expression of immunostimulatory molecules. In mice with conditional targeting of PD-1 in myeloid cells, which also displayed diminished tumor growth, TAMs had gene expression profiles enriched for myeloid differentiation, activation and leukocyte-mediated immunity displaying >50% overlap with enriched profiles of SHP-2-deficient TAMs. In bone marrow, GM-CSF induced the phosphorylation of PD-1 and recruitment of PD-1-SHP-2 to the GM-CSF receptor. Deletion of SHP-2 or PD-1 enhanced GM-CSF-mediated phosphorylation of the transcription factors HOXA10 and IRF8, which regulate myeloid differentiation and monocytic-moDC lineage commitment, respectively. Thus, SHP-2 and PD-1-SHP-2 signaling restrained myelocyte differentiation resulting in a myeloid landscape that suppressed antitumor immunity.

Effect of CYP3A4*22, CYP3A5*3 and POR*28 genetic polymorphisms on calcineurin inhibitors dose requirements in early phase renal transplant patients.

OBJECTIVE: This study aimed to investigate the combined effect of CYP3A5*3, CYP3A4*22, and POR*28 genetic polymorphisms on tacrolimus and cyclosporine dose requirements. METHODS: One hundred thirty renal transplant patients placed on either tacrolimus or cyclosporine were recruited, where the effect of CYP3A5*3, CYP3A4*22, and POR*28 genetic polymorphisms on their dose requirements were studied at days 14, 30, and 90 post-transplantations. RESULTS: The POR*28 allele frequency in the studied population was 29.61%. The tacrolimus dose-adjusted trough concentration ratio (C0/D) was significantly lower in the fast metabolizers group ( CYP3A5*1/POR*28(CT/TT ) carriers) than in the poor metabolizers group ( CYP3A5*3/*3/CYP3A4*22 carriers) throughout the study (14, 30, and 90 days) ( P = 0.001, <0.001, and 0.003, respectively). Meanwhile, there was no significant effect of this gene combination on cyclosporine C0/D. CONCLUSION: Combining the CYP3A5*3, POR*28 , and CYP3A4*22 genotypes can have a significant effect on early tacrolimus dose requirements determination and adjustments. However, it does not have such influence on cyclosporine dose requirements.

Design, synthesis, and apoptotic antiproliferative action of new 1,2,3-triazole/1,2,4-oxadiazole hybrids as dual EGFR/VEGFR-2 inhibitors.

A novel series of 1,2,3-triazole/1,2,4-oxadiazole hybrids (7a-o) was developed as dual inhibitors of EGFR/VEGFR-2. Compounds 7a-o were evaluated as antiproliferative agents with Erlotinib as the reference drug. Results demonstrated that most of the tested compounds showed significant antiproliferative action with GI50 values ranging from 28 to 104 nM, compared to Erlotinib (GI50 = 33 nM), and compounds 7i-m were the most potent. Compounds 7h, 7i, 7j, 7k, and 7l were evaluated as dual EGFR/VEGFR-2 inhibitors. These in vitro experiments demonstrated that compounds 7j, 7k, and 7l are potent antiproliferative agents that may operate as dual EGFR/VEGFR-2 inhibitors. Compounds 7j, 7k, and 7l were evaluated for their apoptotic potential activity, where findings indicated that compounds 7j, 7k, and 7l promote apoptosis by activating caspase-3, 8, and Bax and down-regulating the anti-apoptotic Bcl-2. Molecular docking simulations show the binding mode of the most active antiproliferative compounds within EGFR and VEGFR-2 active sites.

Immunological Signatures in Blood and Urine in 80 Individuals Hospitalized during the Initial Phase of COVID-19 Pandemic with Quantified Nicotine Exposure.

This research analyzes immunological response patterns to SARS-CoV-2 infection in blood and urine in individuals with serum cotinine-confirmed exposure to nicotine. Samples of blood and urine were obtained from a total of 80 patients admitted to hospital within 24 h of admission (tadm), 48 h later (t48h), and 7 days later (t7d) if patients remained hospitalized or at discharge. Serum cotinine above 3.75 ng/mL was deemed as biologically significant exposure to nicotine. Viral load was measured with serum SARS-CoV-2 S-spike protein. Titer of IgG, IgA, and IgM against S- and N-protein assessed specific antiviral responses. Cellular destruction was measured by high mobility group box protein-1 (HMGB-1) serum levels and heat shock protein 60 (Hsp-60). Serum interleukin 6 (IL-6), and ferritin gauged non-specific inflammation. The immunological profile was assessed with O-link. Serum titers of IgA were lower at tadm in smokers vs. nonsmokers (p = 0.0397). IgM at t48h was lower in cotinine-positive individuals (p = 0.0188). IgG did not differ between cotinine-positive and negative individuals. HMGB-1 at admission was elevated in cotinine positive individuals. Patients with positive cotinine did not exhibit increased markers of non-specific inflammation and tissue destruction. The blood immunological profile had distinctive differences at admission (MIC A/B↓), 48 h (CCL19↓, MCP-3↓, CD28↑, CD8↓, IFNγ↓, IL-12↓, GZNB↓, MIC A/B↓) or 7 days (CD28↓) in the cotinine-positive group. The urine immunological profile showed a profile with minimal overlap with blood as the following markers being affected at tadm (CCL20↑, CXCL5↑, CD8↑, IL-12↑, MIC A/B↑, GZNH↑, TNFRS14↑), t48h (CCL20↓, TRAIL↓) and t7d (EGF↑, ADA↑) in patients with a cotinine-positive test. Here, we showed a distinctive immunological profile in hospitalized COVID-19 patients with confirmed exposure to nicotine.

Novel piperine-carboximidamide hybrids: design, synthesis, and antiproliferative activity via a multi-targeted inhibitory pathway.

A new series of piperine-carboximidamide hybrids VIa-k was developed as a new cytotoxic agent targeting EGFR, BRAF, and CDK2. The antiproliferative effect against four cancer cells was investigated against erlotinib. Hybrids VIc, VIf, VIg, VIi, and VIk have the highest antiproliferative activity. Compounds VIc, VIf, VIg, VIi, and VIk inhibited EGFR with IC50 values ranging from 96 to 127 nM. Compounds VIf and VIk had the most potent inhibitory activity as BRAFV600E (IC50 = 49 and 40 nM, respectively) and were discovered to be potent inhibitors of cancer cell proliferation (GI50 = 44 and 35 nM against four cancer cell lines, respectively). Compound VIk, the most effective derivative as an antiproliferative agent, demonstrated potent anti-CDK2 action with an IC50 value of 12 nM, which is 1.5-fold more potent than the reference dinaciclib. Finally, VIc, VIf, and VIk have a high capacity to inhibit LOX-IMVI cell line survival.

Crosstalk of TNF-α, IFN-γ, NF-kB, STAT1 and redox signaling in lipopolysaccharide/d-galactosamine/dimethylsulfoxide-induced fulminant hepatic failure in mice.

Purpose: The clinical study of fulminant hepatic failure is challenging due to its high mortality and relative rarity, necessitating reliance on pre-clinical models to gain insight into its pathophysiology and develop potential therapies. Methods and Results: In our study, the combination of the commonly used solvent dimethyl sulfoxide to the current-day model of lipopolysaccharide/d-galactosamine-caused fulminant hepatic failure was found to cause significantly greater hepatic damage, as indicated by alanine aminotransferase level. The effect was dose-dependent, with the maximum increase in alanine aminotransferase observed following 200 µl/kg dimethyl sulfoxide co-administration. Co-administration of 200 µl/kg dimethyl sulfoxide also remarkably increased histopathological changes induced by lipopolysaccharide/d-galactosamine. Importantly, alanine aminotransferase levels and survival rate in the 200 µl/kg dimethyl sulfoxide co-administration groups were both greater than those in the classical lipopolysaccharide/d-galactosamine model. We found that dimethyl sulfoxide co-administration aggravated lipopolysaccharide/d-galactosamine-caused liver damage by stimulating inflammatory signaling, as indicated by tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) levels. Further, nuclear factor kappa B (NF-kB) and transcription factor activator 1 (STAT1) were upregulated, as was neutrophil recruitment, indicated by myeloperoxidase activity. Hepatocyte apoptosis was also increased, and greater nitro-oxidative stress was noted, as determined based on nitric oxide, malondialdehyde, and glutathione levels. Conclusion: Co-treatment with low doses of dimethyl sulfoxide enhanced the lipopolysaccharide/d-galactosamine-caused hepatic failure in animals, with higher toxicity and greater survival rates. The current findings also highlight the potential danger of using dimethyl sulfoxide as a solvent in experiments involving the hepatic immune system, suggesting that the new lipopolysaccharide/d-galactosamine/dimethyl sulfoxide model described herein could be used for pharmacological screening with the goal to better understand hepatic failure and evaluate treatment approaches.

Production of biodiesel from oleaginous fungal lipid using highly catalytic bimetallic gold-silver core-shell nanoparticle.

AIMS: This study aims to synthesize, characterize and apply gold-silver core-shell nanoparticles (Au@Ag NPs), a nanocatalyst, to maximize biodiesel production from fungal isolate Fusarium solani (FS12) via a transesterification one-step reaction. METHODS AND RESULTS: The Au@Ag NPs structure was examined by UV-vis spectrophotometer, transmission electron microscopy, X-ray diffraction and Fourier transform infrared (FTIR). All devices were used to characterize Au@Ag NPs and confirmed successful synthesis of nanoparticles. Fungal lipid was quantitatively determined by sulfo-phospho-vanillin colorimetric method. Among 15 F. solani isolates obtained from rhizospheric soils of the date palm, F. solani (AF12) was chosen as the highly significant producer that accumulates above 20% lipid. The maximum biodiesel yield was 91.28 ± 0.19%, obtained under the optimum reaction conditions of 3% Au@Ag NPs as nanocatalyst concentration, and 1:20 oil to methanol molar ratio at 70℃ for 30 min. HPLC method was applied for monitoring in situ transesterification reaction. FTIR spectroscopy was used in qualitative analysis of biodiesel by verifying the presence of unique characteristic peaks of diagnostic significance. The quality of the biodiesel produced was confirmed by the high purity of fatty acid methyl esters analysis content up to >99%. CONCLUSIONS: These findings propose the applicability of F. solani (FS12) as a promising isolate to accumulate lipids and biodiesel production as a feedstock. SIGNIFICANCE AND IMPACT OF THE STUDY: The link between nanotechnology and fungi. Au@Ag NPs were synthesized at room temperature, which displayed high catalytic activity for in situ transesterification reaction. Catalytic activity appeared at low temperature, mole ratio and short reaction time. Oleaginous fungi are described as easily grown, have short life cycle, are cost-effective, and they utilized various sources of carbon up to waste and a simplified process to develop scale-up production as well, economic value, opposite the usage of vegetable oils which need for large agricultural areas.

Why a Propofol Infusion Should Be the Anesthetic of Choice for Auditory Brainstem Response Testing in Children.

BACKGROUND: Auditory brainstem response (ABR) testing is considered to be relatively resistant to effects of volatile anesthetics. The impact of newer anesthetics on interpretability of ABR testing is unknown. This study compared sevoflurane versus propofol anesthesia on qualitative interpretability of ABR click-testing in children. METHODS: This prospective double-blind crossover study enrolled children (≤18 years old) receiving general anesthesia for elective ABR testing. All subjects received both sevoflurane and propofol anesthesias in the same ABR testing session. Deidentified ABR data were reviewed by 5 audiologists (blinded to anesthetic and patient) to determine threshold levels for hearing loss. The primary outcome was qualitative interpretability (false positive) of ABR click-testing. RESULTS: Each patient was tested at 4 different intensities in each ear: generating 624 records under each anesthetic, for a total of 1248 records. A few patients were tested at 5 different intensities in a single ear accounting for the additional 11 records, yielding 1259 records. Under sevoflurane anesthesia, 21 of the same patients (37 ears) were identified with abnormal ABR levels consistent with hearing loss (one or both ears). The probability of a patient being diagnosed with hearing "loss" in one or both ears was significantly less with propofol versus sevoflurane anesthesia (mid P =.0312). If patients with bilateral loss are compared, the mid P value is 0.0098. The effect size based on patients was medium to large, with a minimum value of Cohen w = 0.320. CONCLUSIONS: Sevoflurane produced more false positives for hearing loss and suggested more severe hearing loss than propofol. False-positive ABR tests, produced by certain anesthetic agents, can have significant life-long impact and negative psychosocial and developmental implications. Use of the intravenous anesthetic propofol is superior to sevoflurane for ABR testing in children.

New 1,3,4-oxadiazoles linked with the 1,2,3-triazole moiety as antiproliferative agents targeting the EGFR tyrosine kinase.

A series of 1,3,4-oxadiazole-1,2,3-triazole hybrids bearing different pharmacophoric moieties has been designed and synthesized. Their antiproliferative activity was evaluated against four human cancer cell lines (Panc-1, MCF-7, HT-29, and A-549) using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The preliminary activity test displayed that the most active compounds, 6d, 6e, and 8a-e, suppressed cancer cell growth (GI50 = 0.23-2.00 µM) comparably to erlotinib (GI50 = 0.06 µM). Compounds 6d, 6e, and 8a-e inhibited the epidermal growth factor receptor tyrosine kinase (EGFR-TK) at IC50 = 0.11-0.73 µM, compared to erlotinib (IC50 = 0.08 ± 0.04 µM). The apoptotic mechanism revealed that the most active hybrid 8d induced expression levels of caspase-3, caspase-9, and cytochrome-c in the human cancer cell line Panc-1 by 7.80-, 19.30-, and 13-fold higher than doxorubicin. Also, 8d increased the Bax level by 40-fold than doxorubicin, along with decreasing Bcl-2 levels by 6.3-fold. Cell cycle analysis after treatment of Panc-1 cells with hybrid 8d revealed a high proportion of cell accumulation (41.53%) in the pre-G1 phase, indicating cell cycle arrest at the G1 transition. Computational docking of the 8d and 8e hybrids with the EGFR binding site revealed their ability to bind with EGFR similar to erlotinib. Finally, in silico absorption, distribution, metabolism, and excretion/pharmacokinetic studies for the most active hybrids are discussed.

An Electrostatic MEMS Roll-Pitch Rotation Rate Sensor with In-Plane Drive Mode.

In this paper, we presented a novel electrostatic Roll/Pitch MEMS gyroscope with in-plane drive mode and out-of-plane sense mode. The proposed structure is developed based on a tuning fork gyroscope with decoupled sense mass on each tine that control the sense out-of-plane frequency. A multi-height deep reactive ion etching (DRIE) fabrication process was utilized to achieve and enhance decoupling between the drive and sense modes. We presented our design methodology followed by an analytical and finite element (FEM) model. Our experimental results showed a good match between the analytical model and those obtained experimentally, from the drive and sense oscillation frequencies. Our characterization setup used a custom made application specific integrated circuit (ASIC) for characterization and was able to achieve ARW of 0.2 deg/rt-h, a bias instability 5.5 deg/h, and scale factor non-linearity (SFNL) 156 ppm FS.

Impact of a Clinical Pharmacist Intervention Program on the Follow-up of Type-2 Diabetic Patients.

Aims: The primary aim of this current study was to investigate the impact of the clinical pharmacist interventions on glycemic control and other health-related clinical outcomes in patients with type 2 diabetes in Egypt. Methods: A prospective trial was conducted on 100 patients with uncontrolled type 2 diabetes admitted in the diabetes outpatient's clinics. Patients were randomly allocated into the clinical pharmacist intervention group and usual care group. In the intervention group, the clinical pharmacist, in collaboration with the physician had their patients receive pharmaceutical care interventions. In contrast, the usual care group patients received routine care without clinical pharmacist's interference. Results: After 6-month of follow-up, of the average HbA1c and FBG values of the patients in the clinical pharmacist intervention group (HbA1c % from 8.6 to 7.0; FBG (mg/dL) from 167.5 to 121.5) decreased significantly compared to the usual care group patients (HbA1c % from 8.1 to 7.8; FBG (mg/dL) from 157.3 to 155.9) (P < .05). Additionally, the results indicated that mean scores of patients 'diabetes knowledge, medication adherence, and diabetes self-care activities of the patients in the clinical pharmacist group increased significantly compared to the control group (P < .05). Conclusions: The study demonstrated an improvement in HbA1c, FBG, and lipid profile, in addition to self-reported medication adherence, diabetes knowledge, and diabetes self-care activities in patients with type 2 diabetes who received pharmaceutical care interventions. The study outcomes support the benefits and the need to integrate clinical pharmacist interventions in the multidisciplinary healthcare team in Egypt.

Dietary cold-pressed red and black pepper oil mixture enhances growth, carcass, blood chemistry, antioxidant, immunity and caecal pathogens of quails.

This study was carried out to explore the influence of black and red pepper oils (BRPO) as a natural antibacterial agent on growth, carcass, liver and kidney functions, lipid profile, immunity and antioxidant indices of growing quails. A total of 225 Japanese quails (1-week-old) were haphazardly divided into 5 groups, each group consists of 45 unsexed birds with 5 replications (9 birds each). The first group was fed basal diet (control), and the 2nd, 3rd, 4th and 5th groups fed rations containing BRPO (0.4, 0.8, 1.2 and 1.6 g/kg diet respectively). During 1-3 weeks of age, body weight gain (BWG) and feed conversion ratio (FCR) were improved with dietary BRPO. During 3-5 and 1-5 weeks of age, BWG was quadratically increased with BRPO supplementation. Also, a quadratic improvement in FCR was observed with BRPO during the overall period. There were no statistical differences in all carcass parameters due to BRPO treatment. Feeding quails on rations enriched with BRPO was linearly influenced liver functions. The highest BRPO levels (1.2 and 1.6 g/kg diet) minimized plasma content of total cholesterol (TC) and triglyceride (TG) respectively. Malondialdehyde (MDA) was reduced in quails fed diets supplemented with BRPO. In apart from the highest level of BRPO, complement 3 (C3) was increased with increasing BRPO level. Birds fed diets enriched with BRPO exhibited lower colonization of TBC, lactobacilli, coliform, Salmonella and E. coli than those in the control group. It could be concluded that dietary BRPO can positively affect quail growth, liver and kidney functions, antioxidant and immunity parameters, lipid profile and lipid peroxidation as well as pathogenic bacteria.

Replacement of dietary fish oil with plant oils improves the immunological responses and the antioxidant status in Oreochromis niloticus exposed to suboptimal temperature.

Here, we investigated the effects of total dietary fish oil (FO) substitution with plant oil (PO) on hematological indices, immune status, antioxidant activity, IL1ß and TNF-α gene expression, and hypoxia stress resistance in Oreochromis niloticus at suboptimal temperatures. Fish (n = 360) were randomly divided into 12 circular fiberglass tanks (500 L; 3 replicates for each dietary group, 30 fish/replicate, 90 fish/group). The control group was fed a basal diet with FO as the lipid source. The CO, SFO, and LSO groups were fed a basal diet with complete replacement of FO with corn, sunflower, and linseed oils, respectively. After 4 weeks, no effects on hematological indices were observed in fish fed the experimental diets. The LSO and CO groups showed a significant increase in γ globulin levels. The highest levels of non-specific immune parameters, antioxidant activity, and IL-1ß and TNF-α gene expressions were recorded in the LSO group. The LSO group also exhibited good resistance to hypoxia stress. Therefore, the total dietary substitution of FO with PO (especially LSO and CO) is recommended as a valuable strategy to ameliorate the immunosuppressive effects of suboptimal temperatures and enhance the resistance of O. niloticus to hypoxia stress.

Effects of dietary curcumin and acetylsalicylic acid supplements on performance, muscle amino acid and fatty acid profiles, antioxidant biomarkers and blood chemistry of heat-stressed broiler chickens.

The objective was to investigate the effects of dietary curcumin and acetylsalicylic acid (ASA) on the performance and physiological responses of broiler chickens under chronic thermal stress. One hundred and sixty day-old male chicks (Ross 308) were divided equally into 4 groups (each contained 4 replicates). On the day 22 of age and thereafter, the first group (TN) was raised in a thermoneutral condition (23 ±â€¯1 °C), while the second group (HS) was subjected to 8 h of thermal stress (34 °C) and both groups fed the basal diet with no supplements. The third (CR) and fourth (AS) groups were subjected to the same thermal stress conditions and fed curcumin-supplemented diet (100 mg curcumin kg-1 diet) and ASA-supplemented diet (1 g ASA kg-1 diet), respectively. Dietary treatment had a significant effect on ADFI (P = 0.041), average daily gain (P = 0.013) and final body weight (P = 0.001). The curcumin-supplemented had higher values for these measures compared with other experimental groups (P < 0.05). Also, the dietary curcumin supplement significantly increased the carcass yield as compared to the HS group (P < 0.05). Compared with the HS group, the dietary curcumin and ASA supplements decreased the concentration of malondialdehyde in the breast muscles (P = 0.014). Both dietary supplements exhibited a marked ability to restore the serum TAC, Na and K in heat-stressed broiler chickens. The current study reported a remarkable ability of curcumin supplement to restore the concentrations of polyunsaturated fatty acids (PUFA) in the breast muscles of heat-stressed broilers, including α-linolinec acid and Docosahexaenoic acid (P = 0.009 and 0.001, respectively). It could be concluded that supplemental dietary curcumin or ASA enhanced growth performance and antioxidant biomarkers of heat-stressed broilers. Moreover, curcumin might be an effective dietary supplement to alleviate the adverse effect of chronic thermal stress on carcass yield and meat quality.

Impact of terlipressin infusion during and after live donor liver transplantation on incidence of acute kidney injury and neutrophil gelatinase-associated lipocalin serum levels: A randomized controlled trial.

BACKGROUND AND AIM: Acute kidney injury (AKI) with liver transplantation (LT) is not uncommon. Impact of terlipressin infusion on AKI, hemodynamics, and plasma concentration of neutrophil gelatinase-associated lipocalin (NGAL) was studied. METHODS: Patients (n=50) were randomized (NCT02059460, USA) into two equal groups: terlipressin vs Controls. Terlipressin (1-4 µg/kg/h) was administrated for 5 days. Intraoperative transesophageal Doppler for hemodynamic management. Renal functions, peak portal vein blood flow velocity (PPV), and hepatic artery resistive index (HARI) were recorded. Plasma NGAL (pNGAL) was measured baseline, 2 and 24 hours postreperfusion. RESULTS: Hepatitis C virus (HCV) was the main etiology. Age, sex, model of end-stage liver disease (MELD), and renal functions were comparable. Postoperative AKI incidence and NGAL concentrations were comparable (P>.05) between terlipressin and controls groups (44% vs 48% and 112.5±9 vs 93.1±8 ng/mL), respectively, but intraoperative NGAL in both groups increased significantly 2 hours postreperfusion (P<.05). The three NGAL readings were comparable (P>.05) between AKI (n=23) and non-AKI developers (n=27). Mean arterial blood pressure (MAP) was maintained in both groups with less systemic vascular resistance (SVR) fluctuations with terlipressin. Median norepinephrine consumption was lower in terlipressin vs controls (8 vs 12 mg; P=.04). The PPV and HARI were not affected by terlipressin at any stage (P>.05). CONCLUSION: Postliver transplant AKI was not prevented by terlipressin use nor predicted by NGAL levels.

Dosing and efficacy of intranasal dexmedetomidine sedation for pediatric transthoracic echocardiography: a retrospective study.

PURPOSE: We designed this retrospective observational study on the use of α2-agonist dexmedetomidine to determine the optimum intranasal dose to achieve sedation for pediatric transthoracic echocardiography and to identify any dose-related adverse effects. METHODS: Outpatient children aged three months to three years with diverse diagnoses of congenital heart disease, including cyanotic cardiac defects, underwent transthoracic echocardiography under dexmedetomidine sedation. Aerosolized intranasal dexmedetomidine was administered with initial doses ranging from 1-3 µg·kg(-1). A rescue dose of 1 µg·kg(-1) was administered if adequate sedation was not achieved within 45 min following the first dose. The primary study outcome was the achievement of adequate sedation to allow transthoracic echocardiography (TTE) scanning, including subxiphoid and suprasternal probe manipulation. RESULTS: Sedation with intranasal dexmedetomidine for transthoracic echocardiography was successful in 62 of the 63 (98%) patients studied, with an intranasal rescue dose required in 13 (21%) patients. Intranasal doses of dexmedetomidine 2.5-3.0 µg·kg(-1) were required for tolerating TTE probe placement, including subxiphoid and suprasternal manipulation, with minimal response and a 90% success rate. Excluding patients who required a second dose of dexmedetomidine, the mean (standard deviation) time from administration to achieving such sedation (onset time) was 26 (8) min for low-dose (1-2 µg·kg(-1)) dexmedetomidine and 28 (8) min for moderate-dose (2.5-3.0 µg·kg(-1)) dexmedetomidine (P = 0.33). Time from administration of low-dose dexmedetomidine to discharge, including TTE scan time, was 80 (14) min, and it increased with moderate-dose dexmedetomidine to 91 (22) min (P = 0.05). Mild to moderate bradycardia and hypotension were observed, but no interventions were required. CONCLUSION: We found that aerosolized intranasal dexmedetomidine offers satisfactory conditions for TTE in children three months to three years of age with an optimal dose of 2.5-3.0 µg·kg(-1)administered under the supervision of a pediatric cardiac anesthesiologist.

The sedation mindset: philosophy, science, and practice.

PURPOSE OF REVIEW: Medicine is evolving. An increasing influx of medically complex patients coupled with diminishing resources set the stage for substantial challenges in providing safe, effective sedation and analgesia for children requiring medical procedures. This review will discuss the essential components of a successful sedation plan outside of the traditional operating room setting. RECENT FINDINGS: As the discipline of sedation has developed, specialty societies have created and updated guidelines, policies, and statements intended to guide their own practice. There is a lack of consensus among them regarding appropriate targeted depths of sedation, monitoring requirements, definitions of adverse events, resuscitation skills required, and appropriate sedatives used. A transparent, collaborative approach is needed to ensure the sharing of expertise and to encourage evidence-based consistency and safety optimization across venues and specialties. SUMMARY: To meet this need, a multidisciplinary strategy is essential in training, performance of procedures outside of the operating room, and care coordination. To deliver safe, effective care, the sedationist must: perform a targeted presedation assessment; optimize the patient and family prior to sedation; tailor the induction and maintenance to the specific child's condition, needs, and procedure; safely recover the child; and provide a safe plan for postsedation care.

Differential distribution of IL28B.rs12979860 single-nucleotide polymorphism among Egyptian healthcare workers with and without a hepatitis C virus-specific cellular immune response.

The CC genotype of the interleukin (IL)-28B.rs12979860 gene has been associated with spontaneous hepatitis C virus (HCV) clearance and treatment response. The distribution and correlation of an IL28B.rs12979860 single-nucleotide polymorphism (SNP) with HCV-specific cell-mediated immune (CMI) responses among Egyptian healthcare workers (HCWs) is not known. We determined this relationship in 402 HCWs who serve a patient cohort with ~85% HCV prevalence. We enrolled 402 HCWs in four groups: group 1 (n = 258), seronegative aviremic subjects; group 2 (n = 25), seronegative viremic subjects; group 3 (n = 41), subjects with spontaneously resolved HCV infection; and group 4 (n = 78), chronic HCV patients. All subjects were tested for an HCV-specific CMI response using an ex-vivo interferon-gamma (IFNγ) ELISpot assay with nine HCV genotype-4a overlapping 15-mer peptide pools corresponding to all of the HCV proteins. All subjects were tested for IL28B.rs12979860 SNP by real-time PCR. An HCV-specific CMI was demonstrated in ~27% of the seronegative aviremic HCWs (group 1), suggesting clearance of infection after low-level exposure to HCV. The frequency of IL28B.rs12979860 C allele homozygosity in the four groups was 49%, 48%, 49%, and 23%, while that of the T allele was 14%, 16%, 12 and 19%, respectively, suggesting differential distributions among subjects with different HCV status. As reported, IL28B.rs12979860 predicted the outcome of HCV infection (p < 0.05), but we did not find any relationship between the IL28B genotypes and the outcome of HCV-specific CMI responses in the four groups (p > 0.05). The data show differential IL28B.rs12979860 genotype distribution among Egyptian HCWs with different HCV status and could not predict the outcome of HCV-specific CMI responses.

Interleukin 28B.rs12979860 genotype does not affect hepatitis C viral load in Egyptians with genotype 4 chronic infection.

Several host and viral factors affect the natural history of Hepatitis C Virus (HCV) infection. Interleukin 28B (IL28B).rs12979860 single nucleotide polymorphism (SNP) was found to predict viral clearance with and without therapy. Subjects with the CC (favorable) genotype of IL28B.rs12979860 were more likely to spontaneously clear the infection and respond favorably to therapy. These data suggest that subjects with the "favorable" CC genotype might have a lower viral load when compared to those with the "unfavorable" TT genotype. Therefore, we examined the effect of IL28B.rs12979860 SNP on HCV viral load and clearance among HCV-infected Egyptians. This cross sectional study was conducted on 375 HCV antibody-positive subjects. Detection and quantification of HCV-RNA was determined by RT-PCR. IL28B.rs12979860 genotyping was performed using SYBR green real-time PCR and specific primers. Of 375 HCV-antibody positive subjects, 239 (63.7%) had chronic HCV infection while the remaining 136 (36.3%) subjects had spontaneously cleared the virus. The frequency of IL28-B CC, CT, and TT genotypes among spontaneous resolvers were 54.4%, 39.0%, and 6.6% while among the chronically infected subjects, they were 31.4%, 49.8%, and 18.8%, respectively. As expected, IL28 genotype predicted spontaneous HCV clearance (p < 0.001). The average HCV viral loads were 1.5 ± 0.69 x 10(6), 0.62 ± 0.11 x 10(6) and 0.51 ± 0.14 x 10(6) IU/ml among chronic subjects with the IL28B.rs12979860 CC, CT and TT genotypes, respectively (p > 0.05). In conclusion, our results show that IL28B.rs12979860 genotype does not affect viral load among chronic HCV infected Egyptians. These findings further confirm the complexity of viral host interactions in determining HCV infection outcome.

Detection of Aspergillus flavus in stored peanuts using real-time PCR and the expression of aflatoxin genes in toxigenic and atoxigenic A. flavus isolates.

Aspergillus flavus is the main species from section Flavi responsible for aflatoxin accumulation in stored peanuts. Rapid methods to detect A. flavus could help to prevent aflatoxins from entering the food chain. A real-time polymerase chain reaction (RTi-PCR) assay was standardized for rapid, specific, and sensitive detection of A. flavus in stored peanuts. A. flavus was detected in 53.6% and 50% of peanut samples by RTi-PCR and A. flavus and Aspergillus parasiticus agar culture, respectively, with 95% agreement between them. Twenty-two A. flavus isolates were screened using high-performance liquid chromatography for their capacity to produce aflatoxin AFB1 (B1). B1 was produced by >72% of the isolates. Sixteen isolates produced B1 at concentrations ranging from 1.64 to 109.18 µg/mL. Four aflatoxin biosynthetic pathway genes (aflD, aflM, aflP, and aflQ) were evaluated using PCR and reverse-transcription PCR in 22 A. flavus isolates from peanut kernels with the aim of rapidly and accurately differentiating toxigenic and atoxigenic isolates. The PCR amplification of genes did not correlate with aflatoxin production capability. The expression of aflD and aflQ was a good marker for differentiating toxigenic from atoxigenic isolates.