Assessment of the impact of cytomegalovirus seropositivity on blood parameters in renal hemodialysis patients.

End-stage renal disease (ESRD) patients are considered immunocompromised, putting them at high risk for infections, including cytomegalovirus (CMV). CMV can affect hematological parameters, causing further complications in ESRD patients. This study intended to determine the seropositivity of CMV infection in hemodialysis patients and its effect on different blood parameters in ESRD patients to help decrease the overall dialysis associated morbidity and mortality. Blood samples were collected from 45 ESRD patients and 45 controls. A complete blood count was performed using an automated cell counter. CMV-specific IgM and IgG levels were measured using immunochemistry testing. The seropositivity for CMV-IgG was 42.2% in ESRD patients which was significantly higher than in control group (22.2%) (p=0.042). The seropositivity for CMV-IgM was 6.7% in ESRD patients with no difference with the control group (4.4%). The prevalence of anemia was significantly higher in CMV seropositive (77.3%) compared to CMV seronegative (47.8%) ESRD patients. Other studied blood parameters were not different between CMV seronegative and seropositive ESRD patients. In conclusion, CMV infection is a significant concern for dialysis patients and can affect hematological parameters, leading to further complications. Early detection and treatment of CMV infection and monitoring of CMV IgM and IgG levels are critical to prevent further complications and improve clinical outcomes.

Screening for SARS-CoV-2 IgM and IgG antibodies among healthcare workers: A single-center study in Egypt.

Coronavirus disease 2019 (COVID-19) pandemic has become a global public health disaster, spreading throughout the world. In order to accurately determine the extent of the pandemic, it is important to accurately identify the prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection among healthcare workers (HCWs). This study intended to determine the prevalence of SARS-CoV-2 infection among HCWs and examine its correlation with the demographic characteristics of the study participants prior to the implementation of the vaccination campaign. In this cross-sectional study included 431 HCWs from Suez Canal University Hospital in Ismailia, Egypt. Their sera were screened for SARS-CoV-2 antibodies using a one-step novel coronavirus (COVID-19) IgM/IgG antibody test from Artron, Canada. Positive cases were then confirmed using nasal swab real-time reverse transcriptase PCR from Viasure, Spain. Of the 431 study participants, 254 (58.9%) were males and 177 (41.1%) females. The majority of participants, 262 (60.8%), were younger than 30 years old, 150 (34.8%) between 30 and 40 years old, and only 19 (4.4%) older than 40 years old. Out of the total samples, 26 (6%) tested positive for SARS-CoV-2 IgM, while 19 (4.4%) tested positive for both IgM and IgG. The majority of the samples, 386 (89.6%), tested negative for both IgG and IgM. There was no association between the prevalence of SARS-CoV-2 and either sex or age of study participants. In conclusion, during the study period, the prevalence of SARS-CoV-2 infection among healthcare workers at Suez Canal University Hospital in Egypt was relatively low. Additionally, there was no significant correlation observed between the prevalence of positive cases and either age or sex.

Three key genes expression profiling in Egyptian rheumatoid arthritis patients.

Rheumatoid arthritis (RA) is a multi-system autoimmune disease with synovial joints involvement. The triad of autoimmunity, genetics, and environment is the key player in RA pathogenesis. We intended to investigate gene expression of C-C Chemokine Ligand 2 (CCL2), protein tyrosine phosphatase non-receptor type 22 (PTPN22), and Cytotoxic T-lymphocyte associated protein 4 (CTLA-4) in RA patients versus controls, and its correlation with the activity of the disease. The relative expression of PTPN22, CTLA-4, and CCL2 in the peripheral blood of 59 RA patients and 50 controls was determined using RT-PCR. There was a significantly higher median (inter-quartile range) expression of CTLA-4 and CCL2 in RA patients in comparison to controls (P < 0.05). However, in RA patients, PTPN22 expression was significantly lower than in controls (P=0.0001). A weak significant correlation was detected between PTPN22 and either CTLA-4 or CCL2. Also, on comparing RA patients with moderate to severe disease activity versus those who have a mild disease activity, CCL2 was significantly over-expressed (P > 0.05). Thus, in Egyptian RA patients, there was a significant PTPN22 down-expression and greater expression of CTLA-4 and CCL2. Moreover, over-expression of CCL2 in RA patients with moderate-to-severe disease activity was significant. We conclude that these three key genes could become useful diagnostic markers for RA and CCL2 expression as a good prognostic tool for RA disease activity.

Evaluation of blood and biochemical parameters of COVID-19 patients in Suez Canal University Hospital; A retrospective study.

INTRODUCTION: The severe acute respiratory syndrome coronavirus 2 (SARS CoV2-CoV-2) viral outbreak in Wuhan (China) caused thousands of confirmed cases and deaths around the world. Severe viral pneumonia with respiratory failure and death are the ultimate consequence of infection. AIM: This study aimed to evaluate the regularly performed standard laboratory parameters that can assist in COVID-19 case identification and establish an effective approach to help care and management of (COVID-19) patients. METHODOLOGY: COVID-19 (n = 129) patients were hospitalized in the Suez Canal University Hospital and were retrospectively examined. Laboratory parameters were gathered from patients upon admission (n = 129) during the period from the 20th of June to 15th of August 2020. SARS-CoV-2 cases were diagnosed clinically and radiologically by chest Computed Tomography (CT) and confirmed by RT-PCR. RESULTS: The results showed that COVID-19 survivors exhibited lower hemoglobin (Hb) and hematocrit (HCT), while showed higher Red Cell Distribution Width (RDW), neutrophil lymphocyte ratio (NLR), and lymphocytes. Logistic regression analysis showed that age greater than 60 years old, neutrophilia and high NLR were associated with more deaths. CONCLUSION: Monitoring of lymphopenia, neutrophilia and NLR may help categorizing patients who may need Intensive care.

The Association of Angiopoietin-2 1064 C/T Rs3020221 Gene Polymorphism with Knee Osteoarthritis.

Osteoarthritis (OA) is a common type of arthritis, affecting millions of people around the world. Angiopoietin-2 (Angpt-2) has a role in the development of chronic inflammatory diseases. We aimed to assess the serum Angpt-2 levels in knee OA patients and to investigate the association of Angpt-2 gene polymorphism(rs3020221 C/T) with knee OA susceptibility and severity. Angiopoietin-2(rs3020221C/T) gene polymorphism was identified in 254 knee OA patients and 227 healthy controls using real-time polymerase chain reaction. Serum Angpt-2 was measured using ELISA. The Arabic version of the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) Index and Kellgren-Lawrence (KL) grading score were used to assess the clinical and radiological severity of OA and their relationship with Angpt-2(rs3020221C/T) gene polymorphism was investigated. Serum Angpt-2 levels were significantly higher in knee OA patients than in the controls (P = .001). OA patients with C/T genotype had a four times greater risk of developing OA than other genotypes (OR = 4.39, 95% CI = 2.85-6.76). Additionally, the T allele presented more in OA patients 224/508 (44%) with two times risk of developing OA (OR = 1.86, 95% CI = 1.43-2.43, p = .001). Angpt-2 SNP (rs3020221C/T) genotype C/T was significantly associated with elevated serum Angpt-2 levels (14.15 ± 5.62 ng/ml). The serum Angpt-2 levels are significantly elevated in OA patients and Angpt-2 gene polymorphism (rs3020221 C/T) may be a risk factor for OA development and both are associated with the severity of knee OA. Carriers of the C/T genotype have a significantly higher serum Angpt-2 levels and a greater risk of developing OA.

Convalescent plasma in COVID-19: renewed focus on the timing and effectiveness of an old therapy.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the coronavirus disease 2019 (COVID-19) pandemic that has strained health care systems worldwide and resulted in high mortality. The current COVID-19 treatment is based on supportive and symptomatic care. Therefore, convalescent plasma (CP), which provides passive immunization against many infectious diseases, has been studied for COVID-19 management. To date, a large number of randomized and non-randomized clinical trials as well as many systematic reviews have revealed conflicting results. This article summarizes the basic principles of passive immunization, particularly addressing CP in COVID-19. It also evaluates the effectiveness of CP as a therapy in patients with COVID-19, clinical trial reports and systematic reviews, regulatory considerations and different protocols that are authorized in different countries to use it safely and effectively. An advanced search was carried out in major databases (PubMed, Cochrane Library, and MEDLINE) and Google Scholar using the following key words: SARS-CoV-2, COVID-19, convalescent plasma, and the applied query was "convalescent plasma" AND "COVID-19 OR SARS-CoV-2". The results were filtered and duplicate data were removed. Collective evidence indicates that two cardinal players determine the effectiveness of CP use, time of infusion, and quality of CP. Early administration of CP with high neutralizing anti-spike IgG titer is hypothesized to be effective in improving clinical outcome, prevent progression, decrease the length of hospital stay, and reduce mortality. However, more reliable, high quality, well-controlled, double-blinded, randomized, international and multicenter collaborative trials are still needed.

Assessment of human parvovirus B19 infection in Egyptian hemodialysis patients.

INTRODUCTION: Parvovirus B19V has been shown to be associated with end-stage renal disease (ESRD) with increased risk of post-infection anemia, especially in hemodialysis (HD) patients. This effect may be due to immunosuppression, insufficient erythropoietin, or short lifespan of red blood cells. Therefore, parvovirus infection should be investigated in this group of patients suffering from anemia or pancytopenia. We assessed the frequency of parvovirus B19 in HD patients attending Suez Canal University Hospital and analyzed the correlation of this infection with hematological parameters in those patients compared with normal individuals. METHODS: We recruited 80 ESRD patients on hemodialysis and 70 healthy controls. History-taking, full examination, and complete blood count (CBC) were performed for all study subjects. Parvovirus B19 detection was performed through polymerase chain reaction (PCR), which included the QIAamp DNA Mini Kit for extracting DNA, which was amplified using TaqMan Universal Master Mix and detected using TaqMan MGB probes by real-time PCR using Rotor Gene Analyzer (6000). FINDINGS: HD patients had a significantly higher frequency of B19V infection than the control group (p = .02). We also found that parvovirus B19-infected HD patients had significantly lower CBC values than uninfected patients. CONCLUSION: The frequency of parvovirus B19 was significantly higher in HD patients and was associated with lower hematological parameters than in uninfected patients, suggesting a significant role of this virus in the pathogenesis of anemia and/or pancytopenia in ESRD.

Genotyping of Helicobacter pylori Virulence Genes cagA and vacA: Regional and National Study.

Helicobacter pylori (H. pylori) plays a crucial role in the pathogenesis of gastritis, peptic ulcer, and gastric cancer. The presence of pathogenicity islands (PAI) genes contributes to the pathogenesis of many gastrointestinal disorders. Cytotoxin-associated gene A (cagA) and vacuolating cytotoxin gene (vacA) are the most known virulence genes in H. pylori. So, our aim was to study H. pylori virulence genes' role in gastric disorders pathogenesis. Our study included 150 adult patients who suffered dyspeptic symptoms and were referred to the GIT endoscopy unit. Gastric biopsies were attained for rapid urease test (RUT) and histopathological examination, and multiplex PCR technique for detection of virulence genes was performed. It was found that 100 specimens were (RUT) positive, of which sixty samples (60%) were PCR positive for H. pylori ureC gene. The vacA and cagA genes were identified in 61.6% and 53% of H. pylori strains, respectively. Only 5 cases were vacA-positive and cagA-negative. The most virulent vacA s1 allele existed in 56.6% of cases. Out of the 60 H. pylori strains, 66% had at least one virulence gene and 34% did not show any virulence gene. H. pylori infection showed significant increase with age. H. pylori are prevalent amid dyspeptic patients in our region. The main genotype combinations were vacA+/cagA+ of s1m1 genotype and they were frequently associated with peptic ulcer diseases, gastritis, and gastroesophageal reflux disease.

Mental Stress, Anxiety and Depressive Symptoms and Interleuken-6 Level among Healthcare Workers during the COVID-19 Pandemic.

OBJECTIVES: This study aimed to evaluate the interleukin-6 (IL-6) levels and its relationship to stress, anxiety, and depressive symptoms among healthcare workers (HCWs) compared to controls during the COVID-19 pandemic. METHODS: A total of 80 HCWs in Suez Canal University Hospital in Ismailia, Egypt, and 80 controls were analyzed during the COVID-19 pandemic. The Depression, Anxiety and Stress Scale (DASS 21) questionnaire was used, and serum IL-6 level was determined in both groups. RESULTS: IL-6 levels were high in 81.2% (65) of HCWs compared to 36% (45) of controls (P < .05). The DASS score was higher in participants with high IL-6 levels (>3 ng/mL) than in those with mild to moderate levels (P < .05). The regression model revealed that the type of work as a healthcare staff, irregular or night shift, and stress were predictors of increased IL-6 levels among the studied sample (P < .05) (odds ratio = 20.30, 2.44, and 2.04, respectively). CONCLUSION: The IL-6 level and DASS score were higher in HCWs compared to those in controls during the COVID-19 pandemic. The type of work as a healthcare staff, stress, and irregular or night shift were predictors of increased IL-6 levels.

Prevalence of Aminoglycoside Resistance and Aminoglycoside Modifying Enzymes in Acinetobacter baumannii Among Intensive Care Unit Patients, Ismailia, Egypt.

BACKGROUND: Acinetobacter baumannii is an opportunistic pathogen that rapidly develops antibiotic resistance against commonly prescribed antimicrobial agents in hospitalized patients worldwide. Aminoglycosides are commonly used in the treatment of A. baumannii health care-associated infections (HAIs). Aminoglycosides resistance mechanisms are varied and commonly involve production of aminoglycoside-modifying enzymes (AME) and efflux systems. AIM: This study aimed to provide an insight into the frequency of genes encoding AME in A. baumannii strains isolated from different clinical specimens in intensive care units (ICU). METHODOLOGY: A total of 52 multidrug-resistant (MDR) A. baumannii strains were isolated from ICU, Suez Canal University Hospitals. Species identification and antibiotics susceptibility testing were done by the automated system VITEK 2. The genes encoding AME were detected by PCR. RESULTS: Aminoglycosides resistance (amikacin, gentamicin and tobramycin) was observed in 35 isolates (67.3%). We found that aacC1 gene was the predominant AME resistance gene among A. baumannii isolates, detected in 14 isolates (40%), aphA6 in 11 isolates (31.4%) and addA1 in 5 isolates (14.2%). We found 5 isolates containing 2 AME genes, 3 of them with aacC1 and aphA6 and the remaining 2 with both aacC1 and aadA1 genes. Nearly, 5 isolates (14.2%) were negative for all AME resistance genes. CONCLUSION: Our study indicated that AME encoding genes are predominant in A. baumannii strains in our region which stressed on the importance of preventive measures to control spreading of resistance genes.

Amelioration of pulmonary aflatoxicosis by green tea extract: An in vivo study.

Aflatoxins (AFB1) are mycotoxins known to be associated with human and animal diseases. The lung is a at risk from AFB1exposure either via inhalation or circulation. Green tea consumption is increasing over time due to widespread popularity as antioxidants, anti-inflammatory, and cytoprotective agents. Therefore, we attempted to study the lung toxicity caused by AFB1 and the possible ameliorating effect of green tea extract. Forty adult male albino rats were divided into five groups; Group I: Untreated control group, Group II (vehicle): Each rat received 1 ml of olive oil, Group III (GTE): Each rat received Camellia sinensis, green tea extract (30 mg/kg/day), Group IV(AFB1): Each rat received (50 µg/kg/day of AFB1). Group V (AFB1+ GTE): Each rat received the same previously mentioned doses of AFB1 in addition to GTE concomitantly. All treatments were orally gavaged for 8 weeks then rats were sacrificed. Serum levels of pro-inflammatory (IL-1ß, TNF-α, IL-6) and anti-inflammatory (IL-10) cytokines were measured, lung tissues' oxidative stress indices were also measured in addition to the histopathological study which was performed by using hematoxylin & eosin and Masson trichrome stains. Morphometric and statistical analyses were also performed. Oral gavage of AFB1 resulted in significant histopathological changes in the lung tissues, in the form of variable degrees of congestion, hemorrhage, interstitial inflammation with infiltration by chronic inflammatory cells, interstitial fibrosis, bronchitis, vasculitis and fibrous thickening of arterial walls. Inflammation was evident by elevated levels of pro-inflammatory cytokines and a declined level of anti-inflammatory cytokines. Also, oxidative stress was evident by increased levels of Malondialdehyde (MDA), Myeloperoxidase (MPO), and decreased levels of total glutathione (tGSH) and Catalase (CAT). The histopathological changes, inflammatory cytokines, and oxidative stress markers were significantly decreased during concomitant administration of green tea extract in (AFB1+ GTE) group. Aflatoxin B1 has deleterious effects on the lung tissue that could be minimized by concomitant administration of Green tea extract owing to its anti-inflammatory, antioxidant, and protective properties.

Serum level of interleukin-22 in patients with cutaneous warts: A case-control study.

BACKGROUND: Warts are viral cutaneous infections caused by human papilloma virus (HPV), presented by verrucous growth over the skin surface. The immune response is considered to play a crucial role in HPV clearance. It depends on intact cellular immunity including natural killer (NK) cell and cytotoxic T cells. It has been clarified that T-helper (Th) 1 cytokines (interleukin (IL)-2, interferon-γ, and tumor necrosis factor-a) and IL-17 are involved in HPV clearance. IL-22 is one of IL-10 family of cytokines produced by NK cells, Th1, Th17, and Th22 cells. In the skin, IL-22 reduces keratinocyte cornification and enhances keratinocyte production of antimicrobial peptides. IL-22 overexpression has been demonstrated in various viral infections and skin inflammatory disorders. AIM: The aim of this study was to assess serum levels of IL-22 in patients with warts and its association with their different clinical characteristics. METHODS: The study included 20 patients with warts and 20 control subjects. Serum concentration of IL-22 was measured by enzyme-linked immune sorbent assay. RESULTS: Serum levels of IL-22 were significantly higher in patients with warts than in control subjects (P < .001). The levels were significantly higher in patients with recurrent warts after prior treatment than in patients with first-time warts (P = .007). Moreover, a significant positive correlation was detected between serum levels of IL-22 and the number of warts (P = .017). CONCLUSION: Serum level of IL-22 was elevated in patients with warts. Thus, IL-22 may have a crucial role in the antiviral immune response against this infection.

Inducible clindamycin resistance in clinical isolates of staphylococcus aureus in Suez Canal University Hospital, Ismailia, Egypt.

INTRODUCTION: The increasing incidence of methicillin resistance among Staphylococci has led to renewed interest in the usage of macrolide-lincosamide-streptogramin B (MLSB) antibiotics to treat S. aureus infections, with clindamycin being the preferable agent owing to its excellent pharmacokinetic properties. Inducible clindamycin resistance my lead to therapeutic failure. AIM: Detection of the prevalence of constitutive and inducible clindamycin resistance in clinical isolates of S. aureus to improve the clinical outcomes in patients. METHODOLOGY: A total of 176 non-duplicate staphylococcal isolates were isolated from different clinical samples. Methicillin resistance was detected using Cefoxitin disk diffusion (CDD) method. Phenotypic clindamycin resistance was performed for all isolates by D test. Polymerase Chain Reaction (PCR) assay were done for detection of erm resistance genes (ermA, ermB and ermC). RESULTS: Out of 176 strains of S. aureus, 108 isolates (61.3%) were identified as MRSA. Erythromycin and clindamycin resistance was detected in 96 isolates (54.5%) and 68 isolates (38.6%) respectively. Clindamycin resistance (cMLSB) was significantly higher (p value < 0.001) in MRSA strains (56 isolates) compared to MSSA (12 isolates). Resistant genes were detected in 160 isolates (91%). The ermA gene was detected in 28 isolates (16%), the ermB gene was detected in 80 isolates (45.5%) (p < 0.001). CONCLUSIONS AND RECOMMENDATIONS: The frequency of constitutive and inducible clindamycin resistance in MRSA isolates emphasizes the need to use D test in routine antimicrobial susceptibility testing to detect the susceptibility to clindamycin as the inducible resistance phenotype can inhibit the action of clindamycin and affect the treatment efficacy.

Human equilibrative nucleoside transporter-1 and deoxycytidine kinase can predict gemcitabine effectiveness in Egyptian patients with Hepatocellular carcinoma.

BACKGROUND: Several biomarkers of gemcitabine effectiveness have been studied in cancers, but less so in hepatocellular carcinoma (HCC), which is identified as the fifth most common cancer worldwide. Investigation of human equilibrative nucleoside transporter-1 (HENT-1) and deoxycytidine kinase (DCK), genes involved in gemcitabine uptake and metabolism, can be beneficial in the selection of potential cancer patients who could be responding to the treatment. AIM: To study HENT-1 and DCK gene expression in HCC patients with different protocols of treatment. METHODS: Using real-time PCR, we analyzed expression levels of HENT-1 and DCK genes from peripheral blood samples of 109 patients (20 controls & 89 HCC patients) between March 2015 and March 2017. All the 89 HCC patients received the antioxidants selenium (Se) and vitamin E (Vit.E) either alone (45 patients) or in combination with gemcitabine (24 patients) or radiofrequency ablation (RFA) (20 patients). RESULTS: There was a significant increase in HENT-1 expression levels in HCC patients treated with Se and Vit.E alone as compared to controls (P Ë‚ .0001), while there was no significant difference between HCC patients treated with gemcitabine or RFA as compared to controls. In contrast, expression of DCK was significantly increased in all groups of HCC patients as compared to controls (P Ë‚ .0001). CONCLUSIONS: HENT-1 and DCK mRNA expressions are important markers of HCC and for GEM effect and GEM sensitivity in patients with HCC. This could be beneficial in the selection of HCC patients sensitive to gemcitabine to avoid subjecting resistant patients to unnecessary chemotherapy.

Sry-related High Mobility Group Box 17 Functions as a Tumor Suppressor by Antagonizing the Wingless-related Integration Site Pathway.

A transcription factor Sry-related high mobility group box (Sox) 17 is involved in developmental processes including spermatogenesis, cardiovascular system, endoderm formation, and so on. In this article, we firstly review the studies on the relation between the Sox17 expression and tumor malignancy. Although Sox17 positively promotes various tissue development, most of the cancers associated with Sox17 show decreased expression levels of Sox17, and an inverse correlation between Sox17 expression and malignancy is revealed. We briefly discuss the mechanism of such Sox17 down-regulation by focusing on DNA methylation of CpG sites located in the Sox17 gene promoter. Next, we overview the function of Sox17 in the fetal hematopoiesis, particularly in the dorsal aorta in midgestation mouse embryos. The Sox17 expression in hematopoietic stem cell (HSC)-containing intra-aortic hematopoietic cell cluster (IAHCs) is important for the cluster formation with the hematopoietic ability. The sustained expression of Sox17 in adult bone marrow HSCs and the cells in IAHCs of the dorsal aorta indicate abnormalities that are low lymphocyte chimerism and the aberrant proliferation of common myeloid progenitors in transplantation experiments. We then summarize the perspectives of Sox17 research in cancer control.

Maintenance of hematopoietic stem and progenitor cells in fetal intra-aortic hematopoietic clusters by the Sox17-Notch1-Hes1 axis.

The aorta-gonad-mesonephros region, from which definitive hematopoiesis first arises in midgestation mouse embryos, has intra-aortic hematopoietic clusters (IAHCs) containing hematopoietic stem cells (HSCs) and hematopoietic progenitor cells (HPCs). We previously reported expression of the transcription factor Sox17 in IAHCs, and overexpression of Sox17 in CD45lowc-KIThigh cells comprising IAHCs maintains the formation of cell clusters and their multipotency in vitro over multiple passages. Here, we demonstrate the importance of NOTCH1 in IAHC formation and maintenance of the HSC/HPC phenotype. We further show that Notch1 expression is positively regulated by SOX17 via direct binding to its gene promoter. SOX17 and NOTCH1 were both found to be expressed in vivo in cells of IAHCs by whole mount immunostaining. We found that cells transduced with the active form of NOTCH1 or its downstream target, Hes1, maintained their multipotent colony-forming capacity in semisolid medium. Moreover, cells stimulated by NOTCH1 ligand, Jagged1, or Delta-like protein 1, had the capacity to form multilineage colonies. Conversely, knockdown of Notch1 and Hes1 led to a reduction of their multipotent colony-forming capacity. These results suggest that the Sox17-Notch1-Hes1 pathway is critical for maintaining the undifferentiated state of IAHCs.

Thrombopoietin contributes to the formation and the maintenance of hematopoietic progenitor-containing cell clusters in the aorta-gonad-mesonephros region.

In the midgestation mouse embryo, hematopoietic cell clusters containing hematopoietic stem/progenitor cells arise in the aorta-gonad-mesonephros (AGM) region. We have previously reported that forced expression of the Sox17 transcription factor in CD45lowc-Kithigh AGM cells, which are the hematopoietic cellular component of the cell clusters, and subsequent coculture with OP9 stromal cells in the presence of three cytokines, stem cell factor (SCF), interleukin-3 (IL-3), and thrombopoietin (TPO), led to the formation and the maintenance of cell clusters with cells at an undifferentiated state in vitro. In this study, we investigated the role of each cytokine in the formation of hematopoietic cell clusters. We cultured Sox17-transduced AGM cells with each of the 7 possible combinations of the three cytokines. The size and the number of Sox17-transduced cell clusters in the presence of TPO, either alone or in combination, were comparable to that observed with the complete set of the three cytokines. Expression of TPO receptor, c-Mpl was almost ubiquitously expressed and maintained in Sox17-transduced hematopoietic cell clusters. In addition, the expression level of c-Mpl was highest in the CD45lowc-Kithigh cells among the Sox17-transduced cell clusters. Moreover, c-Mpl protein was highly expressed in the intra-aortic hematopoietic cell clusters in comparison with endothelial cells of dorsal aorta. Finally, stimulation of the endothelial cells prepared from the AGM region by TPO induced the production of hematopoietic cells. These results suggest that TPO contributes to the formation and the maintenance of hematopoietic cell clusters in the AGM region.

Interleukin-22 and Chemokine Interferon γ -Inducible-10 (IP-10) Levels in Chronic Hepatitis C Patients and Treatment Response to Pegylated Interferon and Ribavirin Therapy.

IL-22 plays a vital role in improving hepatic damage by targeting hepatocytes that express high levels of IL-22 receptor1. IP-10 is a chemokine that recruit mononuclear cells to liver parenchyma and improves the host immune response against hepatitis C virus. The study targeted 27 patients with chronic HCV who received pegylated Interferon and Ribavirin. IL-22 and IP-10 serum levels were measured by Elisa. The level of the serum IL-22 is higher in HCV patients groups receiving the antiviral treatment compared to control group and its levels significantly increased with response to treatment. The level of the serum IP-10 is higher in HCV patients groups compared to control group and its level significantly decreased with effective antiviral treatment. In conclusion, IL-22 and IP-10 levels could be used with a high sensitivity and specificity during antiviral treatment of HCV infected patients as predictive markers for treatment response.

Sox17 as a candidate regulator of myeloid restricted differentiation potential.

Sry related high mobility group box 17 (Sox17), which is a marker of endodermal cells and a transcriptional regulator, has a critical role in the maintenance of fetal and neonatal hematopoietic stem cells (HSC). Sox17 has been identified as a key regulator of the development and differentiation of fetal hematopoietic progenitors from the aorta-gonad-mesonephros (AGM) region. The co-culture of Sox17-transduced hematopoietic progenitor cells (CD45(low) c-Kit(high) cells) from AGM regions on OP9 stromal cells gives rise to multipotential hematopoietic stem/progenitor cells. Here, we show that in a primary transplantation experiment, Sox17-transduction in CD45(low) c-Kit(high) cells of embryonic day (E) 10.5 AGM increased the absolute number of common myeloid progenitors (CMPs) in the bone marrow (BM) of recipient mice in comparison to that of granulocyte/macrophage progenitors (GMPs) and the megakaryocyte/erythroid progenitors (MEPs). When Sox17-transduced cells were cultured with OP9 stromal cells, Sox17-transduced GMPs (Sox17-GMPs), Sox17-transduced CMPs (Sox17-CMPs), and Sox17-transduced MEPs (Sox17-MEPs) were generated. Sox17-GMPs and Sox17-CMPs maintained their self-renewal capacity and the hematopoietic ability upon co-culture with the OP9 stromal cells for some passages. Moreover, Sox17-GMPs exhibited the increase in expression of c-Mpl and GATA-2 in comparison to GMPs of BM and Sox17-CMPs showed the increase in expression of c-Mpl, NF-E2, and ß-globin genes in comparison to CMPs of BM. Furthermore, when Sox17-transduced cells were cultured in methylcellulose to examine the colony-forming ability, Sox17-GMPs and Sox17-CMPs maintained the formation of mixed colonies for some passages. Taken together, Sox17 is suggested to regulate the maintenance and differentiation of hematopoietic progenitors derived from AGM regions at midgestation, in particular myeloid progenitors.

Sox17-mediated maintenance of fetal intra-aortic hematopoietic cell clusters.

During mouse development, definitive hematopoiesis is first detected around embryonic day 10.5 (E10.5) in the aorta-gonad-mesonephros (AGM) region, which exhibits intra-aortic cell clusters. These clusters are known to contain hematopoietic stem cells (HSCs). On the other hand, it is not clear how the cells in such clusters maintain their HSC phenotype and how they are triggered to differentiate. Here we found that an endodermal transcription factor marker, Sox17, and other F-group (SoxF) proteins, Sox7 and Sox18, were expressed in E10.5 intra-aortic cell clusters. Forced expression of any of these SoxF proteins, particularly Sox17, in E10.5 AGM CD45(low) c-Kit(high) cells, which are the major component of intra-aortic clusters, led to consistent formation of cell clusters in vitro during several passages of cocultures with stromal cells. Cluster-forming cells with constitutive Sox17 expression retained long-term bone marrow reconstitution activity in vivo. Notably, shutdown of exogenously introduced Sox17 gene expression resulted in immediate hematopoietic differentiation. These results indicate that SoxF proteins, especially Sox17, contribute to the maintenance of cell clusters containing HSCs in the midgestation AGM region. Furthermore, SoxF proteins play a pivotal role in controlling the HSC fate decision between indefinite self-renewal and differentiation during fetal hematopoiesis.