Altered microRNA expression profile is linked to T-cell exhaustion-related pathways in pediatric patients with acute lymphoblastic leukemia.

BACKGROUND: Although the phenotype and functions of exhausted T cells in several cancers have been identified, the involved molecular mechanisms remain to be further elucidated. In this regard, we have recently reported that the immunoregulatory cells, including myeloid-derived suppressor cells (MDSCs) and regulatory T cells (Tregs), share common dysregulated miRNAs that target specific immunosuppressive pathways in patients with in acute lymphoblastic leukemia (ALL). AIM: In this study, we aimed to further explore whether similar dysregulation in miRNA expression is linked to T cell exhaustion and dysfunctionality in B cell ALL patients. METHODS: Peripheral blood samples from pediatric patients with ALL were recruited before and after induction chemotherapy as well as from healthy donors. Affymetrix microarray platform was used for miRNA profiling, and qRT-PCR was used to validate the expression of certain miRNAs that are related to T cell exhaustion. Bioinformatics analysis was performed to explore whether the dysregulated miRNAs were linked to T-cell exhaustion related pathways. RESULTS: A total of 516 miRNAs were dysregulated in ALL patients as compared to the healthy donor. Furthermore, among the total analyzed miRNAs, 10 were found to be linked to the key genes implicated in three exhaustion-related pathways; TGF-ß, FOXO, and MAPK, as revealed by miR-pathway analysis. Moreover, qRT-PCR analysis showed similar expression pattern to those obtained by microarray analysis. CONCLUSION: Our pilot study suggests the implication of certain miRNAs in T cell exhaustion pathways via targeting the specific key genes in those pathways.

Relation between Vitamin B12 Levels and Smell Affection in COVID-19 Patients

Abstract Introduction Olfactory and gustative alterations are frequent in the initial stages of the COVID-19 infection. Vitamin B12 deficiency has been linked to olfactory dysfunction. Objective The present study aimed to assess the relationship between vitamin B12 levels and smell affection in COVID-19 patients. Methods The present study included 201 laboratory-confirmed COVID-19 patients. Smell affection was assessed using self-rated olfactory function. Serum vitamin B12 levels were assessed using commercial enzyme-linked immunosorbent assay (ELISA) kits. Results According to the smell function assessment, the patients were classified into three categories: normal osmesis (n = 77), hyposmia (n = 49), and anosmia (n = 75) (►Fig. 1). Four weeks later, 195 patients (97.0%) had their normal smell function restored. The remainder 6 patients included 4 anosmic and 2 hyposmic patients. Patients with hyposmia or anosmia had significantly lower vitamin B12 levels when compared with patients with normal osmesis (median [IQR]: 363.0 [198.0-539.0] versus 337.0 [175.0-467.0] and 491.0 [364.5-584.5] pg/ml, respectively, p < 0.001). Conclusion Vitamin B12 appears to have some contribution to smell affection in patients with COVID-19 infection.

Relation between Vitamin B12 Levels and Smell Affection in COVID-19 Patients.

Introduction Olfactory and gustative alterations are frequent in the initial stages of the COVID-19 infection. Vitamin B12 deficiency has been linked to olfactory dysfunction. Objective The present study aimed to assess the relationship between vitamin B12 levels and smell affection in COVID-19 patients. Methods The present study included 201 laboratory-confirmed COVID-19 patients. Smell affection was assessed using self-rated olfactory function. Serum vitamin B12 levels were assessed using commercial enzyme-linked immunosorbent assay (ELISA) kits. Results According to the smell function assessment, the patients were classified into three categories: normal osmesis ( n = 77), hyposmia ( n = 49), and anosmia ( n = 75) ( Fig. 1 ). Four weeks later, 195 patients (97.0%) had their normal smell function restored. The remainder 6 patients included 4 anosmic and 2 hyposmic patients. Patients with hyposmia or anosmia had significantly lower vitamin B12 levels when compared with patients with normal osmesis (median [IQR]: 363.0 [198.0-539.0] versus 337.0 [175.0-467.0] and 491.0 [364.5-584.5] pg/ml, respectively, p < 0.001). Conclusion Vitamin B12 appears to have some contribution to smell affection in patients with COVID-19 infection.

Prevalence of iron deficiency anemia and beta thalassemia carriers among relatives of beta thalassemia patients in Nile Delta region, Egypt: a multicenter study.

BACKGROUND: Screening of ß thalassemia among close relatives is more feasible in highly prevalent countries with limited resources. The purpose of this study is to determine the prevalence of ß thalassemia carriers and iron deficiency anemia among relatives of ß thalassemia patients in Mid Delta, Egypt. METHODS: This is a cross-sectional multi-center study conducted on 2118 relatives of patients with ß thalassemia from different Egyptian governorates in the Mid Delta region. They were subjected to history taking with precise determination of geographic location, general examination, and the following investigations: complete blood counts, serum ferritin for those who showed microcytic hypochromic anemia, and high-performance liquid chromatography for those who were not diagnosed as iron deficiency anemia. RESULTS: The total prevalence of iron deficiency anemia among close relatives of confirmed ß thalassemia patients in the Nile Delta region was 17.19%. The highest prevalence of iron deficiency anemia (45.05%) was reported in Al-Gharbia Governorate, followed by Al-Menoufia Governorate (21.67%), and the lowest prevalence was that of Al-Sharkia Governorate (4.91%). The differences were highly statistically significant (p < 0.001). ß thalassemia carrier prevalence rate in the studied relatives was 35.84%, with the highest prevalence detected in Al-Sharkia Governorate (51.32%), followed by Kafr-Alsheikh and Al-Dakahilia Governorates (41.78%, 37.13%) respectively, while Al-Menoufia Governorate had the lowest prevalence rate (25.00%). These differences were also highly statistically significant (p < 0.001). CONCLUSION: More than one-third of relatives of patients with ß thalassemia are carriers of the disease, while 17.19% suffer from iron deficiency anemia. This study demonstrates the importance of tracing the high number of beta thalassemia carriers among relatives of patients with ß thalassemia in Egypt.

Correction to: MicroRNA signature in hepatocellular carcinoma patients: identification of potential markers.

The correct spelling of the 7th authors' name is Mona Watany.

MicroRNA signature in hepatocellular carcinoma patients: identification of potential markers.

MicroRNAs (miRNAs) play important roles in liver pathologies and they are potential biomarkers for diagnosis of liver diseases progression. Changes in miRNA sera expression can be used as non-invasive biomarkers for hepatocellular carcinoma (HCC). The aim of the study was to identify the miRNome profiling of HCC and its diagnostic value in distinguishing HCC from healthy individuals. Expression profiles of miRNAs in serum samples of 20 HCC patients and 10 healthy controls were detected. Whole miRNome profiling was done using next generation sequencing. Receiver operating characteristic (ROC) analysis was performed to assess the diagnostic performance of the deregulated miRNAs for discriminating HCC cases from healthy controls. MiRNA 142 was highly expressed in HCC (P value = 0.023) while miRNAs 191, 22, and 126 were higher in the controls (P value = 0.005, 0.034, 0.010 respectively). We have identified 5 novel miRNAs and they were highly expressed in HCC than controls. Analysis of ROC curve demonstrated that these deregulated miRNAs can be used as a reliable biomarker for detection of HCC with high diagnostic accuracy (AUC = 0.93). We have detected a panel of serum miRNAs that can be used as a reliable noninvasive screening biomarker of HCC. The study recommends further research to shed light on a possible role of the newly discovered novel miRNAs in HCC pathogenesis.

Stem Cell Therapy Using Bone Marrow-Derived Mononuclear Cells in Treatment of Lower Limb Lymphedema: A Randomized Controlled Clinical Trial.

BACKGROUND: Up till now, there is no satisfactory treatment for lymphedema. The aim of this study is to evaluate stem cell therapy in lymphedema. METHODS AND RESULTS: This prospective randomized study includes 40 patients with chronic lymphedema divided randomly into two groups: group I (stem cell therapy group) and group II (control group). In group I, bone marrow was aspirated and mononuclear cells were separated and then transplanted into the patients. In group II, patients compression therapy alone was applied. Group I included 20 patients (12 males and 8 females), their age ranged from 18 to 38 years with a mean age of 24.8 ± 6.39 years, whereas group II included 20 patients (10 males and 10 females), their age ranged from 18 to 36 years with a mean value of 25.6 ± 8.18 years. In group I, there was a decrease in the mean circumference at ankle after 6 months, which was statistically significant (t = 3.250, p = 0.014). This was associated with marked improvement of pain and walking ability. Whereas in group II, the change in the circumference was statistically insignificant (t = 1256, p = 0.349) with no satisfactory pain relief and improvement in walking ability. Biopsies examined by immunohistochemistry showed marked increase in the number of lymphatic capillaries in group I. CONCLUSION: Stem cell therapy can achieve improvement in limb circumference as well as pain relief and improvement in walking ability in patients with chronic lymphedema compared with those in control group.

Chemotherapy alters the increased numbers of myeloid-derived suppressor and regulatory T cells in children with acute lymphoblastic leukemia.

INTRODUCTION: Acute lymphoblastic leukemia (ALL) is the most common cancer diagnosed in children. The precise mechanism behind the relapse in this disease is not clearly known. One possible mechanism could be the accumulation of immunosuppressive cells, including myeloid-derived suppressor cells (MDSCs) and T regulatory cells (Tregs) which we and others have reported to mediate suppression of anti-tumor immune responses. AIM: In this study, we aimed to analyze the numbers of these cells in a population of B-ALL pediatric patients. METHODS: Peripheral blood samples withdrawn from B-ALL pediatric patients (n = 45 before, during and after the induction phase of chemotherapy. Using multi parametric flow cytometric analysis. MDSCs were identified as Lin-HLA-DR-CD33+CD11b+; and Treg cells were defined as CD4+CD25+CD127-/low. RESULTS: Early diagnosed B-ALL patients showed significant increases in the numbers of MDSCs and Tregs as compared to healthy volunteers. During induction of chemotherapy, however, the patients showed higher and lower numbers of MDSCs and Treg cells, respectively as compared to early diagnosed patients (i.e., before chemotherapy). After induction of chemotherapy, the numbers of MDSCs and Treg cells showed higher increases and decreases, respectively as compared to the numbers in patients during chemotherapy. CONCLUSION: Our results indicate that B-ALL patients harbor high numbers of both MDSCs and Tregs cells. This pilot study opens a new avenue to investigate the mechanism mediating the emergence of these cells on larger number of B-ALL patients at different treatment stages.

Stem cell therapy as a novel therapeutic intervention for resistant cases of alopecia areata and androgenetic alopecia.

BACKGROUND: Management of alopecia areata (AA) and androgenetic alopecia (AGA) is often challenging as patients may be resistant to currently available modalities of treatment. The use of stem cells may be a novel option for resistant cases. OBJECTIVE: To evaluate the safety and efficacy of the use of autologous bone marrow derived mononuclear cells (including stem cells) as compared to follicular stems cells for the management of resistant cases of AA and AGA. METHODS: This study included 40 patients (20 AA patients and 20 AGA patients), all patients were treated with a single session of intradermal injection of autologous stem cells (SCs) therapy. They were divided into four groups according to the applied modality [either autologous bone marrow derived mononuclear cells (bone marrow mononuclear cells [BMMCs] or autologous follicular stem cells [FSC]). RESULTS: Six months after stem cell therapy (SCT) injection, there was a significant improvement, confirmed by immunostaining and digital dermoscopy. The mean improvement in all groups was "very good". There was no significant difference between both methods in either type of alopecia. No serious adverse events were reported. CONCLUSION: Autologous BMMCs and FSC seem to be a safe tolerable and effective treatment for the management of both resistant AA and AGA.

Autologous transplantation of CD34(+) bone marrow derived mononuclear cells in management of non-reconstructable critical lower limb ischemia.

Patients with a decrease in limb perfusion with a potential threat to limb viability manifested by ischemic rest pain, ischemic ulcers, and/or gangrene are considered to have critical limb ischemia (CLI). Because of this generally poor outcome, there is a strong need for attempting any procedure to save the affected limb. The aim of this work is to evaluate the possibility to use stem cell therapy as a treatment option for patients with chronic critical lower limb ischemia with no distal run off. This study includes 20 patients with chronic critical lower limb ischemia with no distal run off who are unsuitable for vascular or endovascular option. These patients underwent stem cell therapy (SCT) by autologous transplantation of bone marrow derived mononuclear cells. 55 % of patients treated with SCT showed improvement of the rest pain after the first month, 60 % continued improvement of the rest pain after 6 months, 75 % after 1 year and 80 % after 2 years and continued without any deterioration till the third year. Limb salvage rate after STC was 80 % after the first year till the end of the second and third years. SCT can result in angiogenesis in patients with no-option CLI, providing a foundation for the application of this therapy to leg ischemia.

Autologus bone marrow stem cells in atrophic acne scars: A pilot study.

BACKGROUND: Acne scar is a very distressing and difficult problem for physicians and patients. Management of cutaneous scarring from acne can be challenging and confusing. The available modalities may be effective, having considerable morbidity and long downtime. Besides, they may not have the same efficacy in different skin types or acne scar types. OBJECTIVE: To evaluate the short-term safety and efficacy of autologous bone marrow (BM) stem cells (SCs) in treating atrophic acne scars. METHODS: Fourteen patients with moderate to severe atrophic acne scars were included. All patients were subjected to single session of autologous BMSCs therapy. Each patient received 5 µg/kg/day granulocyte colony-stimulating factor (G-CSF) as a single subcutaneous dose for 2 successive days before BM aspiration. The SC-containing solution was injected under each scar intradermally. The scars of the patients were clinically assessed both qualitatively and quantitatively before and after 6 months. The patients were given a preformed questionnaire Cardiff acne disability index (CADI) before and after treatment. RESULTS: After 6 months of the injection, there was significant improvement in the qualitative grading, quantitative grading and CADI scores. All types of scars showed significant improvement. No significant adverse effects were reported in any patient. CONCLUSION: Autologous BMSCs seem to be a safe and effective treatment option for the management of all types of atrophic facial acne scars.

Evaluation of the protective effect of Cystone against cisplatin-induced nephrotoxicity in cancer patients, and its influence on cisplatin antitumor activity.

PURPOSE: Evaluating the role of cystone, a polyherbal preparation, in protecting cancer patients against cisplatin-induced nephrotoxicity, and its impact on the cytotoxic activity of cisplatin. METHODS: A prospective open-label randomized controlled trial conducted on 49 cancer patients who received six cycles of 70 mg/m(2) cisplatin-based regimens. The study comprised two groups, a control group (A) in which 28 patients received cisplatin without cystone supplement, and an experimental group (B) in which 21 patients received cisplatin with cystone supplement. Renal function parameters including serum creatinine, creatinine clearance, blood urea, and serum cystatin C were compared between both groups throughout chemotherapy cycles. Patient response to treatment was evaluated in both groups after 3rd and 6th cycles. RESULTS: At the end of the study, mean levels of serum creatinine, blood urea, and serum cystatin C were significantly lower, whereas creatinine clearance was significantly higher in group (B) compared with group (A). In group (B), there was no significant difference between mean levels of renal markers at baseline and after completion of treatment; while significant changes were observed in group (A). Grading of acute kidney injury according to Common Terminology Criteria for Adverse Events revealed significantly better renal status among patients in group (B) "grades 0 and 1 in 76 and 24 % of the patients, respectively" compared with group (A) "grades 0, 1, and 2 in 36, 32, and 32 % of the patients, respectively". Based on Response Evaluation Criteria in Solid Tumors, there was no significant difference between both groups. CONCLUSIONS: Cystone can protect cancer patients from cisplatin nephrotoxicity without interfering with its antitumor activity.

Prevalence of hepatitis C infection among children with ß-thalassaemia major in Mid Delta, Egypt: a single centre study.

BACKGROUND: Transfusion dependant patients are at a higher risk of acquiring bloodborne infections even under conditions of safe transfusion. This study was designed to determine sero-prevalence of hepatitis C infection and possible associated risk factors in thalassaemic children. METHODS: One hundred and twenty five children with ß thalassaemia major (ß-TM) were recruited from the Haematology/Oncology Unit, Paediatric Department, Tanta University Hospital, Egypt, between April 2010 and October 2011. Patients underwent history taking, full clinical examination, routine investigations and venous blood sampling. Serum was stored at -20°C till tested for hepatitis C (HCV Ab) and B (HBsAg) by ELISA. HCV Ab positive cases were confirmed by PCR. RESULTS: All patients were HBsAg negative. HCV Ab ELISA was positive in 76%, negative in 20% and equivocal in 4%. Fifty patients (40%) had positive PCR for HCV. PCR showed low viraemia in 78%, moderate viraemia in 20% and high viraemia in 2%. A positive family history of HCV, history of minor operative intervention and/or dental procedures were significantly associated with higher frequency of HCV infection in thalassaemic children, while amount and frequency of transfused blood, age at transfusion and chelation state were not. CONCLUSION: HCV infection is highly prevalent in children with ß-TM in Egypt despite strict pre-transfusion blood testing. This should arouse the attention for environmental and community acquired factors. Quality management to insure infection control in minor operative procedures and adding more sensitive tests for blood screening are recommended.

Is mycophenolate mofetil superior to pulse intravenous cyclophosphamide for induction therapy of proliferative lupus nephritis in Egyptian patients?

BACKGROUND: Recent studies have suggested that mycophenolate mofetil (MMF) may offer advantages over intravenous cyclophosphamide (IVC) for the treatment of lupus nephritis. The aim of this study was to evaluate the efficacy of MMF compared with IVC in the induction therapy of proliferative lupus nephritis. METHODS: We randomly assigned 47 patients with newly diagnosed active proliferative lupus nephritis class III or IV to open-label oral MMF 2 g/day for 6 months or intravenous cyclophosphamide 0.5-1 g/m(2) monthly for 6 months in addition to corticosteroids. RESULTS: In the intention-to-treat analysis, 14 of the 24 patients (58.33%) receiving MMF and 12 of the 23 patients receiving cyclophosphamide (52.17%) had remission (P = 0.48); complete remission occurred in 6 of the 24 patients (25%) and 5 of the 23 patients (21.74%), respectively (P = 0.53). Improvements in packed cell volume, the erythrocyte sedimentation rate, anti-double-stranded DNA antibodies titer (anti-dsDNA), serum complement, proteinuria, urinary activity, renal function, serum soluble interleukin-2 receptor alpha concentration and the systemic lupus activity measure score were similar in both groups. Two patients assigned to MMF and another patient assigned to IVC developed end-stage renal failure with commencement of dialysis. Adverse events were similar. Major infections occurred in two patients in each group. There was no difference in gastrointestinal side effects, but more diarrhea occurred in those receiving MMF. CONCLUSION: In this 24-week trial, MMF or IVC combined with corticosteroids demonstrated equal efficacy in inducing remission of proliferative lupus nephritis.

Quantitative measurement of telomerase reverse transcriptase mRNA and chromosomal analysis of urine by M-FISH in the diagnosis and follow-up of bladder cancer.

This study was performed to evaluate the clinical utility of the measurement of the expression of telomerase enzyme (the catalytic subunit of the complex hTERT) and of the chromosomal analysis of urine by multicolor fluorescence in situ hybridization (M-FISH) assay for the detection of bladder cancer and its recurrence. These results were compared with those afforded by urine cytology, hematuria screening and the bladder tumor antigen (BTA) and fibrin degradation products (FDP) tests. Urine samples were obtained from three groups: 30 patients with bladder cancer, 15 patients with non-malignant bladder disorders and 8 healthy individuals. hTERT mRNA was measured by reverse transcription real-time PCR. M-FISH was performed using a mixture of fluorescent labeled probes for the centromeric regions of chromosomes 3, 7, 17, and the locus specific identifier p16 probe for the 9p21 locus. We demonstrated that the overall sensitivity of hemoglobin dipstick and the BTA and FDP tests was insignificantly greater than that of urine cytology, but with a lower specificity. The hTERT mRNA expression marker offered significantly greater sensitivity for detecting all bladder cancers, especially superficial and low grade tumors, than did urine cytology, and with a higher specificity. M-FISH was significantly more sensitive than urine cytology in detecting bladder tumors, but had lower specificity (insignificant results). The superior sensitivity of M-FISH was maintained when comparing the two assays in terms of low-stage and low grade tumor detection. M-FISH was the most sensitive and specific test in detecting tumor recurrence, followed by the BTA test. hTERT, hematuria screening and FDP showed relatively low sensitivity and low specificity during follow-up. Our findings suggest that the assessment of hTERT expression and chromosomal abnormalities in urine represent reliable tools - equally specific yet far more sensitive than conventional cytology - for the early detection of bladder cancer. The high sensitivity of FISH in detecting recurrence makes it useful for reducing the number of cyctoscopies usually performed in the accurate follow-up of these cases.

Study of gene expression of CD30 variant (CD30v) and CD30 ligand (CD30L) in acute leukemia.

Proliferation of malignant lymphohematopoietic cells is thought to be regulated by a number of surface molecules on tumour cells whose expression may contribute to neoplastic transformation. In this work, reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the gene expression (mRNA) of CD30 variant (CD30v) and CD30 Ligand (CD30L) on the peripheral blood mononuclear cells (PBMCs) of 15 healthy individuals as a control group, 15 patients with newly diagnosed acute myeloid leukemia (AML) and 15 patients with newly diagnosed acute lymphocytic leukemia (ALL). The results revealed that simultaneous positive expression of both CD30v and CD30L was found in 46.7%, 40% and 53.3% of whole leukemic patients and those with AML and ALL respectively, with significant difference from controls in whom no expression was found (P=0.007, 0.021 and 0.005, respectively). Patients with positive expression of CD30v and CD30L were found to have significantly increased blast cell % (p<0.001), increased total leucocytic count (P<0.001) and decreased platelets count (P<0.001) than those with negative expression. No significant difference in expression could be noticed in relation to age (p>0.05), sex (P=0.998.) or hemoglobin (Hb) level (P=0.20). As regard to immunophenotypes of ALL, positive expression was found to be significantly higher in B-cell than T-cell subtype (77.8% versus 16.7%, P=0.02). It could be concluded that frequent expression of CD30V and CD30L was detected only in newly diagnosed cases of AML and ALL, but not in healthy individuals. Positive expression was also significantly associated with more aggressive disease and with B-cell than T-cell subtypes. These results suggested a possible role of these molecules in pathogenesis of such hematopoietic malignancy. Further studies are needed for better understanding of the involved mechanisms.

Incidence of MLL rearrangement in acute myeloid leukemia, and a CALM-AF10 fusion in M4 type acute myeloblastic leukemia.

To determine the incidence of the mixed lineage leukemia (MLL) gene rearrangements in acute myeloid leukemia (AML) without cytogenetically-detected 11q23 abnormalities, we screened 64 cases of AML at diagnosis for MLL rearrangement by FISH. Three cases (4.7%) had a MLL rearrangement detected; one was shown to have a cryptic t(11;22)(q23;q11) and another to have a t(9;11)(p21-22;q23) which had been missed by the conventional cytogenetic study. No 11q23 structural abnormality was visible in the third case. Twenty-six of the 64 cases were further studied by Southern blotting and DNA hybridization, and four of these cases (15%) were found to have MLL rearrangement: in three of these, FISH had not detected any abnormality. FISH was also used to confirm MLL involvement in eight cases of AML that had a cytogenetic abnormality at 11q23; in one of these, Southern blot did not show a rearrangement. The survival of patients with MLL abnormalities identified by cytogenetics, FISH and/or DNA analysis was significantly worse than that of patients without MLL abnormalities (event-free survival p = 0.016) although two patients with a t(9;11)(p21-22;q23) were long-term survivors, consistent with this particular translocation having a better prognosis. One further case with a cytogenetic abnormality close to 11q23 was studied; it was found to have a t(10;11)(p13;q21), and the breakpoints were shown by FISH to involve the Clathrin Assembly Lymphoid Myeloid (CALM) gene at 11q21 and the AF10 gene at 10p13. Our data confirm the value of combining cytogenetic, FISH and molecular analyses to define the incidence and precise nature of MLL and 11q23 abnormalities in AML.