Plasma tissue factor pathway inhibitor levels in coronavirus disease 2019 patients: a systematic review and meta-analysis.

Studies have suggested a relationship between tissue factor pathway inhibitor (TFPI) and coronavirus disease 2019 (COVID-19) severity. However, there is inconsistency in the findings of the studies. To enhance comprehension of this relationship, a meta-analysis was conducted. PubMed, Web of Science, and Scopus databases were searched to identify eligible studies. The mean difference was employed as effect measures and the standardized mean difference (SMD) and the 95% confidence interval (CI) were utilized as a summary statistic. Heterogeneity was assessed through the application of the chi-square test and the I2 statistic. The included studies' quality and risk of bias were assessed using the Newcastle-Ottawa assessment scale, adapted for case-control studies. A total of six studies were included with 684 cases and healthy controls (180 healthy controls and 504 COVID-19 patients with different severity, 76 mild, 292 moderate, and 136 severe). The analysis revealed a significant increase in the TFPI level in COVID-19 patients with moderate severity compared with healthy controls (SMD = 0.95 ng/ml, 95% confidence interval (CI) 0.27, 1.63 ng/ml; I2 : 87.2%). The increased TFPI level in mild and moderate COVID-19 was not significant, SMD = 0.68 ng/ml, 95% CI -0.64 to 2.0 ng/ml; I2 92.9% and SMD = 0.62 ng/ml, 95% CI -0.62 to 1.86 ng/ml; I2 91.5%, respectively. In addition, most studies indicate an association of the increased TFPI concentrations with increased markers of inflammation, endothelial damage, and hypercoagulation. Considering the anticoagulant and anti-inflammatory roles of TFPI, its increase seems to be aimed at modulating COVID-19-induced hyper-inflammation and hyper-coagulation state. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42023437353.

Natural Anticoagulant Protein Levels in Patients With Beta-Thalassemia Major: A Case-Control Study.

Background: ß-thalassemia is a group of inherited blood disorders that affect the production of ß-globin chains, leading to the reduction or absence of these chains. One of the complications observed in patients with ß-thalassemia major (ß-TM) is thrombosis, especially in those who receive frequent blood transfusions. This may be due to a decrease in the levels of the natural anticoagulants: protein C (PC), total protein S (PS), and antithrombin (AT). Methods: In this case-control study, patients with ß-TM, who had received at least 20 packed cell transfusions during their lifetime, were included. Patients with other underlying diseases like bleeding or thrombotic disorders were excluded. Totally, 118 patients with ß-TM and 120 healthy individuals were included. Results: The mean level of PC and AT was significantly lower in patients with ß-TM (48.2 ± 65.4 and 57.42 ± 13.6, respectively) compared to the control group (97.1 ± 21.46 and 81.79 ± 14.3, respectively), with P value of 0.001 and 0.01, respectively. Although the difference was not statistically significant (P = 0.1), a similar trend was observed for total PS (61.12 ± 21.12 for patients versus 72.2 ± 35.2 for the control group). Of note, the decrease in PC, AT, and total PS levels compared to the control group was 50.36%, 27.5%, and 15.34%, respectively. Conclusions: It seems that ß-TM patients who receive prolonged blood transfusions frequently are at an increased risk of decreased in natural anticoagulants levels and therefore potentially are at risk of thrombosis.

Preparation, characterization, and ex-vivo evaluation of curcumin-loaded niosomal nanoparticles on the equine sperm quality during cooled storage.

Oxidative damage to sperm during cooled storage is a significant issue, and curcumin, with its antioxidant properties, could be a solution. However, its low bioavailability presents a challenge that this study aims to address. The primary objective of this study was to investigate the potential of curcumin-loaded niosomal nanoparticles (Cur-LNN) to enhance the antioxidant properties of curcumin and its effect on sperm quality during 72 h cooled storage. The thin-film hydration procedure was applied to prepare Cur-LNN. The fabricated noisomal nanocarriers were characterized using dynamic light scattering (DLS), transmission electron microscopy (TEM), field emission scanning electron microscopy (FESEM), and Fourier-transform infrared (FT-IR) spectroscopy. Moreover, the encapsulation and loading efficiency, in vitro release study, and ex-vivo antioxidant functionality of Cur-LNN on the stallion sperm preserved under cooled storage conditions were assessed. The fabricated Cur-LNN was spherical in shape and had an average particle size of 163.1 ± 1.8 nm, a zeta potential of -34.1 ± 1.9 mV, a poly-dispersity index of 0.339 ± 0.045, an encapsulation efficiency of 92.34 ± 0.18 %, and a loading efficiency of 35.57 ± 1.36 %. Ex-vivo evaluation revealed that supplementation of the semen extender with Cur-LNN has the potential to enhance sperm quality by improving total and progressive motility, plasma membrane functionality, and lipid peroxidation. These results demonstrate that Cur-LNN exhibited stronger antioxidant and protective effects than curcumin. Although further in vivo investigations are warranted, our ex-vivo results suggest that Cur-LNN has the potential to attenuate oxidative damage and can be used to fortify the antioxidant capacity of equine semen under cooled storage conditions.

COVID-19 vaccine-induced immune thrombotic thrombocytopenia: pathophysiology and diagnosis.

Coronavirus disease-19 (COVID-19) vaccine-induced immune thrombotic thrombocytopenia (VITT) is a rare but serious clinical condition with high mortality rate in apparently healthy individuals without noticeable risk factors. VITT typically arises due to the administration of vaccines that possess recombinant adenoviral vectors, including ChAdOx1 nCov-19 (AstraZeneca) and Ad26 COV2.S (Johnson & Johnson/Janssen). Thrombosis frequently occurs at atypical sites, such as the cerebral or splanchnic circulations, in this particular pathological state. Similar to heparin-induced thrombotic thrombocytopenia (HITT), it seems that the cause of VITT is the misdirection of anti-platelet factor 4 antibodies (anti-PF4 Abs), an ancient antimicrobial mechanism. Anti-PF4 Abs in patients with VITT activates the coagulation system, leading to thrombosis. This process occurs through the stimulation of platelets (Plts) and neutrophils and subsequently release of neutrophil extracellular traps (NETs). Due to the potentially fatal consequences of VITT, early diagnosis is mandatory. In addition to thrombocytopenia, thrombosis, and the presence of anti-PF4 Abs, the day of symptoms onset and the elevation of D-dimer are also required for definitive diagnosis of VITT. The absence of one or more criteria can result in the exclusion of definitive VITT and lead to the diagnosis of probable, possible, or unlikely VITT.

An Investigation on Cognitive and Behavioral Effects of Donepezil on Autistic Children in Winter 2020 in Gorgan's Taleghani Pediatric Hospital.

Objectives: The present investigation has been done to study the behavioral effects of donepezil in autistic children, given that not much research has been carried out concerning using this drug for treating autism. Materials & Methods: A cross-sectional and analytic-descriptive study was done on twenty patients with autism, aged 4-17, who visited the neurology clinic of Gorgan's Taleghani Pediatric Hospital and Yasha Pediatric Autism Clinic, Iran from 2019 to 2020. Demographic information, including sex, age, father's education, mother's education ,patient's education, family status, other problems, and ethnicity, were documented using a checklist, having been filled in during interviews with the parents. Before the trial, ABC cognitive and behavioral tests were taken to determine the children's current status. After the tests, these children received a daily dose of donepezil (10mg) before sleep for three months. At the end of the three months, the cognitive and behavioral tests were taken from the children once again. In order to analyze the effects of different factors on the studied variables, including irritability, lethargy, stereotypic behavior, hyperactivity, and inappropriate speech before and after the administration of donepezil in patients, a generalized linear model and to test the effects of donepezil on the studied variables, paired t-test was used. Results: In this study, twenty patients were registered for the investigation, 12 (60%) male and eight (40%) female. Age groups 5-6 had the highest frequency, and age group 17 had the lowest. Regardingthe parents' education, the highest frequency was for bachelor's degrees, and the lowest was for less-than-high school education and master's degree. The highest frequency for the patients' education was kindergarten (60%), and then groups without education (20%) and elementary school-level education (15%). Most of the studied patients (80%) did not have other problems besides autism, and only 20% had other problems besides autism. The family status of 15% of the families was 'separated,' and ethnically, most patients (80%) were Fars, while the rest (20%) were Turkmen. None of the analyzed factors (age, sex, father's education, mother's education, patient's education, other problems, family status, and ethnicity) had a significant effect on the studied variables after the administration of donepezil. Among the studied variables prior to the administration of donepezil and among the analyzed factors, the father's education, the patient's education, other problems, and family status had only a significant effect on stereotypic behavior. The present research findings of the present research indicated that all the studied variables, including irritability, lethargy, stereotypic behavior, hyperactivity, and inappropriate speech, were significantly decreased toward the desired goal. The decreased amounts in irritability, lethargy, stereotypic behavior, hyperactivity, and inappropriate speech after the administration of donepezil were, respectively, 38%, 44%, 54%, 41%, and 54% and on average, these behaviors were reduced by 46%. Conclusion: The present investigation has shown that all the studied variables, including irritability, lethargy, stereotypic behavior, hyperactivity, and inappropriate speech, were significantly decreased towards the desired goal by 46%. This significant decrease is indicative of the effectiveness of the treatment of autism patients with donepezil, and therefore, this drug can be placed as a prominent and essential part of the medical therapy of autism.

Status of major hemostatic components in the setting of COVID-19: the effect on endothelium, platelets, coagulation factors, fibrinolytic system, and complement.

The coagulation, fibrinolytic, anticoagulation, and complement systems are in delicate balance with the vessel wall endothelium ensuring appropriate hemostasis. Coagulopathy in coronavirus disease 2019 (COVID-19) is not a simple disorder of one hemostatic component but a complicated process affecting most of the hemostasis system. COVID-19 disturbs the balance between the procoagulant systems and the regulatory mechanisms. Here, we investigate the effect of COVID-19 on key hemostatic components, including platelets, endothelial cells, coagulation factors, fibrinolytic system, anticoagulant protein system, and complement system, to improve our understanding of the pathophysiological processes underlying COVID-19 coagulopathy based on evidence.

Factor VII Padua in Iran: clinical and laboratory findings of three unrelated patients.

The congenital factor VII (FVII) deficiency with an estimated incidence of one per 300 000 is the most common rare congenital bleeding disorder. The heterogeneous clinical pictures, including asymptomatic to life-threatening manifestations, are seen in patients with FVII deficiency. A variety of gene variants throughout the FVII ( F7 ) gene have been reported so far. In this setting, very rare FVII Padua polymorphism provokes an interesting condition in which results of prothrombin time and FVII activity are different based on the thromboplastin sources used in these tests. The current study aimed to report the phenotype and genotyping of patients with Padua variant. During the workup of the laboratory for FVII deficiency for diagnosis of FVII Padua, all patients with FVII deficiency who had prolonged prothrombin time, normal activated partial thromboplastin time, and variable FVII activity results using different sources of thromboplastin were included. Demographic data and clinical findings were recorded. For the molecular study, the F7 gene sequencing was performed using the Sanger sequencing technique. Five patients with FVII Padua and a history of mild-to-moderate bleeding, including easy bruising, epistaxis, gingivorrhagia, and bleeding after surgical challenges (including dental extraction and tonsillectomy), were detected during the study. DNA sequencing revealed a heterozygote CGG to CAG (Arg364Gln) variant in exon 9 at nucleotide position 1091, consistent with the genetic variant of FVII Padua. Timely diagnosis of FVII Padua is vital to avoid unnecessary exposure of patients to replacement therapy.

Post-COVID-19 splenic infarction in a patient with chronic atrial fibrillation: A case report.

We describe splenic infarction (SI), an infrequent condition, in an 82-year-old COVID-19 patient with chronic atrial fibrillation (AF). COVID-19 may cause thrombosis, and AF is a predisposing factor for splenic infarction. Suspicion of SI may be warranted in COVID-19 patients with abdominal pain, especially if a predisposing factor exists.

Anti-proliferative activity of disulfiram through regulation of the AKT-FOXO axis: A proteomic study of molecular targets.

Due to its potent anti-tumor activity, well-investigated pharmacokinetic properties and safety profile, disulfiram (DSF) has emerged as a promising candidate for drug repurposing in cancer therapy. Although several molecular mechanisms have been proposed for its anti-cancer effects, the precise underlying mechanisms remain unclear. In the present study, we showed that DSF inhibited proliferation of cancer cells by inducing reactive oxygen species (ROS) production, a G1 cell cycle arrest and autophagy. Moreover, DSF triggered apoptosis via suppression of the anti-apoptotic protein survivin. To elucidate the mechanisms for the anti-proliferative activities of DSF, we applied a 2-DE combined with MALDI-TOF-MS/MS analysis to identify differentially expressed proteins in breast cancer cells upon treatment with DSF. Nine differentially expressed proteins were identified among which, three candidates including calmodulin (CaM), peroxiredoxin 1 (PRDX1) and collagen type I alpha 1 (COL1A1) are involved in the regulation of the AKT signaling pathway. The results of western blot analysis confirmed that DSF inhibited p-AKT, suggesting that DSF induces its anti-tumor effects via AKT blockade. Moreover, we found that DSF increased the mRNA levels of FOXO1, FOXO3 and FOXO4, and upregulated the expression of their target genes involved in G1 cell cycle arrest, apoptosis and autophagy. Finally, DSF potentiated the anti-proliferative effects of well-known chemotherapeutic agents such as arsenic trioxide (ATO), doxorubicin, paclitaxel and cisplatin. Altogether, these findings provide mechanistic insights into the anti-growth activities of DSF.

Association between Toxoplasma gondii exposure and hematological malignancies: A systematic review and meta-analysis.

The possible association between Toxoplasma gondii infection and hematological malignancies has been suggested by several studies. The current systematic review and meta-analysis was directed to further understand this relationship. In the present study, five electronic databases were reviewed for T. gondii infection in patients with blood cancer. The random effects model and 95% confidence intervals (CI) were used to determine the overall odds ratio (OR). Heterogeneity was deliberate with Cochran's Q test and I2 statistic. In total, 13 studies including 1504 patients with hematological neoplasia and 2622 subjects without any malignancies were included in this meta-analysis. Overall, 324 patients with blood cancer and 391 subjects without any malignancies were exposed to Toxoplasma infection. The pooled random effect favored a statistically significant increased risk of T. gondii infection in patients with hematological neoplasia compared with non-cancer individuals [OR = 2.43; 95% CI: 1.49-3.97; χ2 = 49.7, I2 = 75.9%, P = 0.00]. Also, significant pooled ORs of positive association were observed for studies which measured anti- Toxoplasma antibodies (IgG) [OR = 2.66; 95% CI: 1.46-4.83; χ2 = 40.3; I2 = 82.6%, P = 0.00]. T-value and P-value were obtained 0.20 and 0.85 by Harbords modified regression test. This meta-analysis demonstrates that toxoplasmosis may be associated with an elevated risk of hematological malignancies. Also, it has found that immunoglobulin class and types of blood cancer are the specific sources of heterogeneity. Additional studies should be performed to examine the impact of T. gondii infection in the onset or development of hematologic neoplasms in the future.

Disulfiram/copper causes ROS levels alteration, cell cycle inhibition, and apoptosis in acute myeloid leukaemia cell lines with modulation in the expression of related genes.

The majority of acute myeloid leukaemia (AML) patients will die from their disease or therapy-related complications. There is an inevitable need to improve the survival of AML patients. Previous studies show that disulfiram (DSF), an anti-alcoholism drug with a low toxicity profile, demonstrates anticancer behaviors. Here, we evaluated the cytotoxicity and mechanistic action of DSF on the AML cell lines KG-1, NB4, and U937. The microculture tetrazolium test revealed that DSF alone or in combination with copper (Cu) is highly toxic to the AML cells at concentrations lower than those achievable in the clinical setting, with Cu increasing the DSF-induced inhibition of metabolic activity. Flow cytometric analysis and QRT-PCR indicated that in the two cell lines, NB4 and U-937, DSF/Cu increased reactive oxygen species (ROS) levels in association with the induction of superoxide dismutase 2 (SOD2) expression and suppression of catalase (CAT). In the KG-1 cell line, DSF/Cu reduced the ROS levels in agreement with the induction of CAT expression. The cell cycle and apoptosis assessment by flow cytometry demonstrated that DSF/Cu induced G0/G1 cell cycle arrest and apoptosis. These were associated with the increased expression of FOXO tumor suppressors, decreased expression of the MYC oncogene and the modulation of their known target genes related to the cell cycle and apoptosis. Therefore, DSF/Cu caused the disturbance of the ROS balance, cell cycle arrest and apoptosis in AML cells in coordination with the modulation in expression of their related genes. These results propose the possible use of DSF in AML therapies.

TGF-ß and Th17 cells related injuries in patients with sulfur mustard exposure.

Sulfur mustard (SM) is a vesicating agent that has been employed as a chemical warfare agent. High-dose exposure to sulfur mustard may lead to the damage of rapidly proliferating cells of bone marrow and, therefore, suppression of the immune system. This may be continued as dysfunction of the immune system, and ultimately result in secondary immune disorders. Studies have suggested a role for T cells in SM-induced lung injury. Moreover, observations from animal studies indicate a delayed-type hypersensitivity (DTH) response after skin exposure to SM, providing an understanding that SM can stimulate specific T cell-mediated immune responses. On the other hand, T helper (Th) 17 cells, which are a subset of CD4+ T cells, have recently been reported to be involved in a number of inflammatory, autoimmune, and chronic fibrotic lung diseases. Furthermore, a strong association has been established between the overproduction of profibrotic cytokines like transforming growth factor (TGF)-ß and Th17 cell number. In this review, we aimed to go through the new findings about the involvement and interactions of TGF-ß and Th17 in SM-related injuries.

Redistribution of cell cycle by arsenic trioxide is associated with demethylation and expression changes of cell cycle related genes in acute promyelocytic leukemia cell line (NB4).

PML-RARα perturbs the normal epigenetic setting, which is essential to oncogenic transformation in acute promyelocytic leukemia (APL). Transcription induction and recruitment of DNA methyltransferases (DNMTs) by PML-RARα and subsequent hypermethylation are components of this perturbation. Arsenic trioxide (ATO), an important drug in APL therapy, concurrent with degradation of PML-RARα induces cell cycle change and apoptosis. How ATO causes cell cycle alteration has remained largely unexplained. Here, we investigated DNA methylation patterns of cell cycle regulatory genes promoters, the effects of ATO on the methylated genes and cell cycle distribution in an APL cell line, NB4. Analysis of promoter methylation status of 22 cell cycle related genes in NB4 revealed that CCND1, CCNE1, CCNF, CDKN1A, GADD45α, and RBL1 genes were methylated 60.7, 84.6, 58.6, 8.7, 33.4, and 73.7%, respectively, that after treatment with 2 µM ATO for 48 h, turn into 0.6, 13.8, 0.1, 6.6, 10.7, and 54.5% methylated. ATO significantly reduced the expression of DNMT1, 3A, and 3B. ATO induced the expression of CCND1, CCNE1, and GADD45α genes, suppressed the expression of CCNF and CDKN1A genes, which were consistent with decreased number of cells in G1 and S phases and increased number of cells in G2/M phase. In conclusion, demethylation and alteration in the expression level of the cell cycle related genes may be possible mechanisms in ATO-induced cell cycle arrest in APL cells. It may suggest that ATO by demethylation of CCND1 and CCNE1 and their transcriptional activation accelerates G1 and S transition into the G2/M cell cycle arrest.

Accuracy of prediction of simulated polygenic phenotypes and their underlying quantitative trait loci genotypes using real or imputed whole-genome markers in cattle.

BACKGROUND: More accurate genomic predictions are expected when the effects of QTL (quantitative trait loci) are predicted from markers in close physical proximity to the QTL. The objective of this study was to quantify to what extent whole-genome methods using 50 K or imputed 770 K SNPs (single nucleotide polymorphisms) could predict single or multiple QTL genotypes based on SNPs in close proximity to those QTL. METHODS: Phenotypes with a heritability of 1 were simulated for 2677 Hereford animals genotyped with the BovineSNP50 BeadChip. Genotypes for the high-density 770 K SNP panel were imputed using Beagle software. Various Bayesian regression methods were used to predict single QTL or a trait influenced by 42 such QTL. We quantified to what extent these predictions were based on SNPs in close proximity to the QTL by comparing whole-genome predictions to local predictions based on estimates of the effects of variable numbers of SNPs i.e. ±1, ±2, ±5, ±10, ±50 or ±100 that flanked the QTL. RESULTS: Prediction accuracies based on local SNPs using whole-genome training for single QTL with the 50 K SNP panel and BayesC0 ranged from 0.49 (±1 SNP) to 0.75 (±100 SNPs). The minimum number of local SNPs for an accurate prediction is ±10 SNPs. Prediction accuracies that were based on local SNPs only were higher than those based on whole-genome SNPs for both 50 K and 770 K SNP panels. For the 770 K SNP panel, prediction accuracies were higher than 0.70 and varied little i.e. between 0.73 (±1 SNP) and 0.77 (±5 SNPs). For the summed 42 QTL, prediction accuracies were generally higher than for single QTL regardless of the number of local SNPs. For QTL with low minor allele frequency (MAF) compared to QTL with high MAF, prediction accuracies increased as the number of SNPs around the QTL increased. CONCLUSIONS: These results suggest that with both 50 K and imputed 770 K SNP genotypes the level of linkage disequilibrium is sufficient to predict single and multiple QTL. However, prediction accuracies are eroded through spuriously estimated effects of SNPs that are distant from the QTL. Prediction accuracies were higher with the 770 K than with the 50 K SNP panel.

Cloning and Expression of Iranian Turkmen-thoroughbred Horse Follicle Stimulating Hormone in Pichia pastoris.

BACKGROUND: Follicle stimulating hormone (FSH) plays an essential role in reproductive physiology and follicular development. OBJECTIVE: A new variant of the equine fsh (efsh) gene was cloned, sequenced, and expressed in Pichia pastoris (P. pastoris) GS115 yeast expression system. MATERIALS AND METHODS: The full-length cDNAs of the efshα and efshß chains were amplified by reverse transcription polymerase chain reaction (RT-PCR) using the total RNA isolated from an Iranian Turkmen-thoroughbred horse's anterior pituitary gland. The amplified efsh chains were cloned into the pPIC9 vector and transferred into P. pastoris. The secretion of recombined eFSH using P. pastoris expression system was confirmed by Western blotting and immunoprecipitation (IP) methods. RESULTS: The DNA sequence of the efshß chain accession number JX861871, predicted two putative differential nucleotide arrays, both of which are located in the 3'UTR. Western blotting showed a molecular mass of 13 and 18 kDa for eFSHα and eFSHß subunits, respectively. The expression of desired protein was confirmed by protein G immunoprecipitation kit. CONCLUSIONS: eFSH successfully expressed in P. pastoris. These findings lay a foundation to improve ovulation and embryo recovery rates as well as the efficiency of total embryo-transfer process in mares.

Silibinin induces apoptosis and inhibits proliferation of estrogen receptor (ER)-negative breast carcinoma cells through suppression of nuclear factor kappa B activation.

BACKGROUND: Silibinin is a traditionally well-known drug for its hepatoprotective efficacy against various types of liver afflictions. In addition, it has recently been considered broadly as a potential chemopreventive agent against many types of cancers. The current study was designed to evaluate the restrictive effects of pharmacological doses of silibinin on SKBR3, an ErbB2-overexpressed and ER-negative human breast carcinoma cell line. METHODS: Effect of silibinin on metabolic activity and proliferation of human breast carcinoma (SKBR3) cell line were evaluated by MTT and BrdU assays respectively. Furthermore, the proapoptotic effect of silibinin was investigated using flow cytometry. The NF-κB phosphorylation assay was also used to assess the effect of silibinin on NF-κB activation. The alkalizing effect of silibinin on SKBR3 cell line was evaluated by measuring pH of media of the silibinin-treated cells compared to control. RESULTS: Our results indicate that silibinin inhibited metabolic activity and cell proliferation of SKBR3 cells in a dose-dependent manner. Moreover, silibinin significantly induced apoptosis in SKBR3 cells. On the other hand, silibinin significantly inhibited activation of NF-κB which is known to be highly active in this cell line. Alkalizing effect of silibinin was also observed. CONCLUSION: The results obtained here indicate that silibinin may be an efficacious therapeutic agent against ER-negative breast carcinomas with high inhibitory effect on NF-κB.

Differential sensitivity of p44/p42-MAPK- and PI3K/Akt-targeted neuroblastoma subtypes to arsenic trioxide.

PI3K/Akt and MAPK/ERK pathways are differentially activated in neuroblastoma (NB) cell types. In an effort to enhance the effectiveness of the NB treatment, we designed experiments to evaluate the effects of ATO in combination with PI3K and MEK1/2 specific inhibitors, LY29004 and U0126, respectively, in SK-N-MC and SK-N-BE(2) cell lines. The results indicated that specific inhibition of PI3K and MEK1/2 significantly enhanced antiproliferative and proapoptotic effects of ATO in SK-N-BE(2), but not in SK-N-MC. Furthermore, in SK-N-BE(2), NF-κB activation was significantly suppressed by LY29004+ATO treatments as compared with ATO alone, indicating that inhibition of PI3K may enhance anti-neoplastic properties of ATO in I-type NB cells through suppression of NF-κB. Moreover, expressions of c-Myc, Bad, Bax and ATM in SK-N-BE(2) cell line were significantly increased by U0126+ATO treatment as compared to treatment with ATO alone. Expression of telomerase hTERT was almost depleted by U0126+ATO treatment. Regarding the fact that activation of PI3K and MAPK in SK-N-BE(2) is higher than in other NB subtypes, we hypothesize that growth of SK-N-BE(2) cell line is highly dependent on these pathways and inhibition of these pathways may has promise for the treatment of multi-drug resistant I-type NB cells by ATO. However, for successful strategies for the treatment of this heterogeneous tumor, other combinations approaches need to be considered to simultaneously target other NB cells.

Azidothymidine hinders arsenic trioxide-induced apoptosis in acute promyelocytic leukemia cells by induction of p21 and attenuation of G2/M arrest.

To enhance anticancer efficacy of the arsenic trioxide (ATO), the combination of ATO and azidothymidine (AZT), with convergence anti-telomerase activity, were examined on acute promyelocytic leukemia (APL) cell line, NB4. In spite of an induction of apoptosis by both drugs separately and a synergistic effect of them on hTERT down-regulation and telomerase inhibition, the ATO-induced cytotoxicity was reduced when it was used in combination with AZT. AZT attenuated the ATO effects on viability, metabolic activity, DNA synthesis, and apoptosis. These observations, despite the deflection from the main goal of this study, dedicate an especial opportunity to elucidate the importance of some of the mechanisms that have been suggested by which ATO induces apoptosis. Cell cycle distribution, ROS level, and caspase-3 activation analyses suggest that AZT reduced the ATO-induced cytotoxic effect possibly via relative induction and diminution of cells accumulated in (G1, S) and (G2/M) phase, respectively, as well as through attenuation of ROS generation and subsequent caspase-3 inhibition. QRT-PCR assay revealed that induction of p21expression by the combined AZT/ATO compared to ATO alone could be a reason for the relative decline of cells accumulation in G2/M and the increase of cells in G1 and S phases. Therefore, the G2/M arrest and ROS generation are likely principle mediators for the ATO-induced apoptosis and can be used as a guide to design rational combinatorial strategies involving ATO and agents with G2/M arrest or ROS generation capacity to intensify ATO-induced apoptosis.

Effect of dietary nutrient density and vitamin premix withdrawal on performance and meat quality of broiler chickens.

BACKGROUND: The objective of this study was to evaluate the effects of feeding high- and low nutrient density diets, and three different vitamin premix withdrawal regimes on broiler performance and meat quality. Male broiler chicks (480 days old) were reared on the floor in a 2 × 3 factorial arrangement for 42 days. Chickens were slaughtered at 42 days of age and meat samples kept at -20 ± 1°C and analysed after 1, 90 and 180 days of storage. RESULTS: Broiler performance was significantly affected by dietary nutrient density. Vitamin premix withdrawal had no significant effect on body weight. The results showed no significant differences between nutrient density and vitamin premix withdrawal on lightness (L*), redness (a*) and yellowness (b*). Oxidative stability of thigh muscle lipids during frozen storage was significantly affected by nutrient density, while vitamin premix withdrawal had no significant impact on lipid oxidation. High nutrient density diet led to a significantly (P < 0.05) decreased pH compared with the low nutrient density diet. CONCLUSION: Increasing dietary nutrient density improved broiler performance but impaired meat quality while vitamin premix withdrawal during finisher periods had no negative effect on broiler performance and meat quality.