Toll-Like Receptor 4, 2, and Interleukin 1 Receptor Associated Kinase4: Possible Diagnostic Biomarkers in Myelodysplastic Syndrome Patients.

Background: Myelodysplastic syndrome (MDS) is a clonal hematologic disorder that requires the integration of morphologic, cytogenetic, hematologic, and clinical findings for a successful diagnosis. Trying to find ancillary tests such as biomarkers improve the diagnosis process. Several studies showed that a disordered immune system is associated with MDS. The chronic activated innate immune system, particularly the Toll-like receptors (TLRs) pathway could be involved in the induction of the inflammation. Materials and Methods: In the present study, we investigated the expression of TLR2, TLR4, and IRAK4 in bone marrow (BM) of MDS patients, the leukemia group, and the healthy group. For this purpose, we assessed the expression of TLR2, TLR4, and IRAK4 by real time-PCR. Results: In line with new findings, we demonstrated that the expression of TLR2, TLR4, and IRAK4 significantly increased in MDS BM compared with the healthy group. Moreover, IRAK4 expression raised significantly in MDS patients compared with other studied hematologic neoplasms. Also, the expression levels of TLR2 and TLR4 significantly increased in MDS in comparison to some studied non-MDS malignancies (P ˂ 0.05). Receiver operating characteristics (ROC) analysis and area under the curve (AUC) suggested that the expression of TLR2, TLR4, and IRAK4 (AUC = 0.702, AUC = 0.75, and AUC = 0.682, respectively) had acceptable diagnostic values to identify MDS from the other understudied leukemias. Conclusion: Overall, the expression of TLR2, TLR4, and IRAK4 could be potential biomarkers for discriminating MDS from some hematologic disorders.

Potential role of serum vitamin D as a risk factor in pediatric acute lymphoblastic leukemia.

Vitamin D deficiency/insufficiency (VDD, VDI) is common in children yet limited experience exists on the association of VDD and hematologic malignancies amongst this population. Therefore, this study aimed to compare serum vitamin D levels in children with acute lymphoblastic leukemia (ALL) and controls. Moreover, vitamin D levels is compared in subjects with and without relapse and evaluated as a prognostic factor for relapse-free survival (RFS). Children with newly diagnosed ALL were recruited as case group. Data on demographic variables as well as the dietary habits were collected by interview. In addition, serum 25(OH)D3 was measured. The case group was followed up for 36 months to assess RFS. Overall, 358 subjects were included in the study (n = 169 cases, n = 189 controls). The mean levels of 25(OH)D3 were 28.05 ± 18.87 and 28.76 ± 12.99 in cases and controls, respectively (p = .68). VDD was found in 15.4% (n = 26) and 4.2% (n = 8) of the case and control groups, respectively (p < .001). Relapse was seen in 18.34% of patients and vitamin D levels of 20 ng/mL or above were associated with longer RFS (p = .044 by log-rank test). In this study, VDD and VDI amongst children with ALL were significantly higher than controls. In addition, lower levels of Vitamin D were associated with increased risk of relapse.

Environmental Risk Factors for Pediatric Acute Leukemia: Methodology and Early Findings.

Background: Acute leukemia is the most common type of malignancy in children, and no major environmental risk factors have been identified relating to its pathogenesis. This study has been conducted with the aim for identifying risk factors associated with this disease. Methods: This study was conducted in 2016-2020 among children aged <15 years residing in Isfahan Province, Iran. Children with newly diagnosed Acute lymphoblastic leukemia, including Acute myeloid leukemia (ALL and AML) were considered a case group. The control group was selected among children hospitalized in orthopedic and surgery wards in the same region. Demographic data, parental occupational exposures and educational level, maternal obstetric history, type of feeding during infancy and parental smoking habits, exposure to pesticides, and hydrocarbons besides dietary habits (using a food frequency questionnaire) were evaluated. Results: Overall, 497 children (195 cases and 302 controls) completed the survey. In the initial analysis, there was no significant difference between case and control groups about type of milk feeding (P = 0.34) or parental age (P = 0.56); however, an association between mothers' education and increased risk for ALL was observed (P = 0.02). Conclusions: The results of this study can be helpful in better understanding the environmental risk factors involved in the incidence of acute leukemia. Future publications based on the analysis of the database created in the present study can lead to recognizing these factors. In addition, evaluating the effect of these factors on treatment outcomes is an important step in reducing the burden of the disease.

Regenerative potential of multinucleated cells: bone marrow adiponectin-positive multinucleated cells take the lead.

BACKGROUND: Polyploid cells can be found in a wide evolutionary spectrum of organisms. These cells are assumed to be involved in tissue regeneration and resistance to stressors. Although the appearance of large multinucleated cells (LMCs) in long-term culture of bone marrow (BM) mesenchymal cells has been reported, the presence and characteristics of such cells in native BM and their putative role in BM reconstitution following injury have not been fully investigated. METHODS: BM-derived LMCs were explored by time-lapse microscopy from the first hours post-isolation to assess their colony formation and plasticity. In addition, sub-lethally irradiated mice were killed every other day for four weeks to investigate the histopathological processes during BM regeneration. Moreover, LMCs from GFP transgenic mice were transplanted to BM-ablated recipients to evaluate their contribution to tissue reconstruction. RESULTS: BM-isolated LMCs produced mononucleated cells with characteristics of mesenchymal stromal cells. Time-series inspections of BM sections following irradiation revealed that LMCs are highly resistant to injury and originate mononucleated cells which reconstitute the tissue. The regeneration process was synchronized with a transient augmentation of adipocytes suggesting their contribution to tissue repair. Additionally, LMCs were found to be adiponectin positive linking the observations on multinucleation and adipogenesis to BM regeneration. Notably, transplantation of LMCs to myeloablated recipients could reconstitute both the hematopoietic system and BM stroma. CONCLUSIONS: A population of resistant multinucleated cells reside in the BM that serves as the common origin of stromal and hematopoietic lineages with a key role in tissue regeneration. Furthermore, this study underscores the contribution of adipocytes in BM reconstruction.

Evaluation of the Relative Frequency of Epstein-Barr Virus Infection in Patients with Recurrent Breast Cancer Compared with Patients with Nonrecurrent Breast Cancer.

Background: The roles of Epstein-Barr virus (EBV) in breast cancer and breast lymphoma by transfecting EBV DNA have been indicated in different studies, but few investigations have been conducted on its roles in recurrence of breast cancer. Here, we aimed to evaluate the roles of EBV in recurrent breast cancer tissue. Materials and Methods: This is a cross-sectional retrospective study that was performed in 2020-2021 in Isfahan on patients with breast cancer. The study population consisted of 30 tissue samples from recurrent breast cancer and 30 samples from nonrecurrent breast cancer. We collected demographic data of patients including age using a checklist. Other collected data were type of cancer, stages of cancer, tumor size in greatest dimension, lymph node involvements, and presence of metastasis. Furthermore, we evaluated all of the pathology samples from both groups for the presence of DNA of EBV and compared the data of both groups. Results: The DNA of EBV was positive in 8 patients of the relapsed group (26.6%) and 7 patients in the nonrelapsed patients (23.3%). There was no significant difference between two groups regarding positive DNA of EBV (P = 0.39). There were no significant differences between two groups of positive DNA of EBV with and without recurrent breast cancer regarding type of cancer (P = 0.63), stage of cancer (P = 0.19), tumor size in greatest dimension (P = 0.31), mean lymph node involvement (P = 0.27), number of lymph node involvement (P = 0.43), and metastasis (P = 0.69). Conclusion: EBV might have no significant role in recurrence of breast cancer.

The prognostic significance of hematogones in childhood B-cell acute lymphoblastic leukemia.

BACKGROUND: Recent studies have demonstrated hematogones (HGs) expansion to be associated with favorable outcomes in hematological diseases, especially in patients with acute myeloid leukemia and patients undergoing hematopoietic stem cell transplantation. Acute lymphoblastic leukemia (ALL) is the most common form of cancer in children. As of now, minimal residual disease (MRD) remains the most compelling independent prognostic factor in childhood ALL. There is need for more prognostic tools for evaluating relapse risk. PROCEDURE: The goal of this study was to assess the prognostic value of HGs on relapse-free survival (RFS) and overall survival (OS) in childhood ALL. In this prospective cohort study, a total of 122 subjects with definitive diagnosis of precursor B lymphoblastic leukemia were evaluated. Flow cytometric HG detection was performed in bone marrow aspirates after induction and consolidation therapy. RESULTS: The median follow-up period of patients was 35.5 ± 9.4 (SD) months. Patients who had at least 1.0% HGs had a significantly better RFS (p = .023). Moreover, univariate and multivariate analyses confirmed that positive HGs were independently associated with longer RFS (unadjusted model: hazard ratio = 0.33, 95% CI = 0.12-0.91, p = .031; adjusted model: hazard ratio = 0.30, 95% CI = 0.11-0.82, p = .020). CONCLUSIONS: Along with the role of MRD, our study shows the significance of HGs as an independent prognostic factor. The results indicate the independent prognostic value of HGs on RFS after adjustment for other prognostic factors, and can be beneficial for risk stratification and treatment modifications amongst pediatric B-cell ALL patients.

Evaluation of CD30 expression in B ALL and its correlation with MRD(Minimum Residual Disease).

BACKGROUND: This study was conducted to evaluate CD30 expression in minimum residual disease after chemotherapy in B-acute lymphoblastic leukemia (B-ALL). MATERIALS AND METHODS: This was a cross-sectional study on 30 new cases of B-ALL between 2018 and 2019. We checked CD30 expressions in fresh bone marrow aspirates by flow cytometry. After 28 days of routine chemotherapy, we calculated minimal residual disease in CD30 positive and negative patients and compare them by Kolmogorov-Smirnov test. RESULTS: Thirty patients with B-ALL with a mean age of 15.62 ± 20.488 were included in the study. CD30 marker was positive in about 10 patients and was negative in about 20 participants. Mean blast count in baseline in CD30 positive group was 77 ± 7.88%, in negative group was 76.3 ± 17.78 % (P = 0.292). After 28 days of chemotherapy mean minimal residual disease (MRD) was 1.07 ± 3.754 in the negative group, 0.12 ± 0.034 in the positive group (P = 0.025). CONCLUSION: Lower MRD on day 28 after chemotherapy was seen in B-ALL patients with baseline CD30 expression.

T-cell immunophenotyping in COVID-19 pneumonia.

The COVID-19 epidemic is currently a global threat that has affected many parts of the world. Some patients require intensive care unit admission due to severe symptoms in the course of the disease. The severity of symptoms in this disease varies from person to person. The effectiveness of the immune response against viral infections depends on the number and activity of T-cells, which play an important role in eliminating virus-infected cells. In this study, we report two patients with COVID-19 pneumonia, one with moderate symptoms and the other with severe symptoms. Although a decrease of absolute lymphocyte count was seen in both patients, a more significant decline reported in the ICU-admitted patient. Expression of activated markers, HLA-DR, CD38, on CD8-positive T-cells was shown in a patient with more severe disease. On the other hand, partial loss of CD7 in the severe case was also observed. Hence, besides of the above parameters that already mentioned in other studies, loss of pan T-markers could be considered as a potentially valuable test for predicting disease severity. We suggest evaluating the predictability of these tests in COVID-19 in larger studies. This study was approved by the Ethics Committee of Isfahan University of Medical Sciences (IR.MUI.MED.REC.1399.238).

Association of lymphocyte subsets with mortality in severe COVID-19 pneumonia patients.

BACKGROUND: Few studies have investigated the alterations in the T and B cell counts and related subgroups in pulmonary infections especially COVID-19. Here, we aimed to evaluate total T and B lymphocytes and T cell subgroup counts to find the possible correlation between number of these cells and severity and mortality in COVID-19 patients. METHODS: This study was performed on 40 patients with severe COVID-19 infection confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and chest HRCT in August 2020. By the time of admission, T lymphocytes profile in peripheral blood was investigated using multicolor flow cytometry. The total number of T lymphocytes, CD4+ T cells, CD8+ T cells, and B lymphocytes were calculated. Expression of CD2, CD3, CD5, and CD7 as pan T cell surface markers and expression of CD38 and HLA-DR as activated markers on T lymphocytes were also evaluated. RESULTS: Nine patients (22.5%) died during the study and 16 patients (40%) were admitted to ICU. Deceased patients demonstrated lower amounts of T cell count and CD4+ T cell count (with a marginal difference (p = 0.07)) compared with survived patients at the time of admission. The chance of mortality was significantly higher for patients with CD7 loss (OR = 14.89). A marginally significant relationship was also indicated between CD4<200/ml and mortality (OR = 8.65), but no other significant relationships were observed between variables and ICU admission. CONCLUSION: Altogether, CD7 loss on T lymphocytes and CD4+ T cell count below 200/ml revealed a significant relationship with mortality. Considering T lymphocytes and T cell subgroup count could have a predictive value for patients suffering from COVID-19.

Blood cell morphology and COVID-19 clinical course, severity, and outcome.

COVID-19 infection affects different organs of the human body, and blood cells are not an exception. Peripheral blood smear (PBS) is a simple and available method to investigate blood cells' morphologic changes. In this study, we aimed to determine the morphologic changes and abnormalities of COVID-19 patients and their relation to the patients' clinical course. In this prospective cross-sectional study, we included 89 PCR-positive COVID-19 patients. A pathologist examined the PBS findings of these patients. The patients' clinical course, including severity, outcome, intubation, and ICU admission, was extracted from their profiles. The statistical analyses were done to find out the relation between PBS findings and patients' clinical course. Results showed that smudge cells are the most frequent abnormality in our participants. Other findings were schistocyte; atypical lymphocytes; and increased large granular lymphocytes, shift to left of granulocytes, giant platelets, and leukoerythroblastic reaction. Our results did not show any statistically significant relationship between PBS findings and their clinical course. Although other studies suggested PBS as a possible predictive tool for COVID-19 disease, our study showed that these findings could not predict nor relate to the patients' clinical course. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12308-021-00459-3.

Prevalence of latent tuberculosis infection and HIV among people who inject drugs in Iran.

INTRODUCTION: Iran has an human immunodeficiency viruses (HIV) epidemic that is concentrated among people who inject drugs (PWID), who have higher risks of progression from latent tuberculosis infection (LTBI) to active disease. The aim of this study is to measure prevalence of LTBI, HIV infection and any risk behaviors among PWID in Iran. METHODS: The cross-sectional study was conducted from August to December 2013 in six cities across Iran. A total of 420 PWID were recruited from drop-in centres using convenience sampling. Trained interviewers collected data on socio-demographic characteristics, drug use history and drug-related risk behaviors across the study sites. A tuberculin skin test (TST) was performed, and HIV infection was assessed by a rapid test. Multivariable modified Poisson regression and logistic regression were used for data analysis. RESULTS: Prevalence of positive TST and HIV positivity was 35.7% and 8.6%, respectively. The prevalence of LTBI and HIV was significantly different across the cities of this study. Positive TST was independently associated with older age (APR 1.03, 95% CI 1.01, 1.05) and being HIV positive (APR 1.89, 95% CI 1.45, 2.47). HIV infection was associated with lifetime history of sharing syringes (AOR 3.28, 95% CI 1.44, 10.71) and lifetime number of imprisonment (AOR 1.09, 95% CI 1.03-1.14). DISCUSSION AND CONCLUSIONS: Prevalence of LTBI infections among PWID is high and independently associated with HIV infection. Given that there are currently no TB services available within drop-in centres, programs which integrate TB case finding, TB preventive therapy, referral and care services for PWID are urgently needed.

COVID-19 and Substance Use Disorders: Recommendations to a Comprehensive Healthcare Response. An International Society of Addiction Medicine Practice and Policy Interest Group Position Paper.

Coronavirus Disease 2019 (COVID-19) is escalating all over the world and has higher morbidities and mortalities in certain vulnerable populations. People Who Use Drugs (PWUD) are a marginalized and stigmatized group with weaker immunity responses, vulnerability to stress, poor health conditions, high-risk behaviors, and lower access to health care services. These conditions put them at a higher risk of COVID-19 infection and its complications. In this paper, an international group of experts on addiction medicine, infectious diseases, and disaster psychiatry explore the possible raised concerns in this issue and provide recommendations to manage the comorbidity of COVID-19 and Substance Use Disorder (SUD).

Angiogenesis and Minimal Residual Disease in Patients with Acute Myeloid Leukemia.

Background: Acute myeloid leukemia (AML) is the most prevalent acute leukemia in adults. Bone marrow angiogenesis is crucial for pathogenesis of leukemia, and increasing bone marrow Mean Vascular Density (MVD) and level of angiogenesis factors are seen in patients with AML. Higher level of bone marrow MVD is associated with poor prognosis of AML according to previous studies. The present study aimed to compare bone marrow MVD in AML patients and controls and evaluate the relation between bone marrow MVD and number of residual blast cells after AML treatment. Materials and Methods: This study is a longitudinal study on AML patients who were admitted to Omid hospital. The bone marrow biopsies of patients with AML and patients with normal diagnosis -as control group- were taken from archives of pathology laboratory. Immunohistochemistry staining was used for all specimens by using thrombomodulin markers for calculating MVD. Flow cytometry findings of AML patients were assessed for percent of minimal residual disease (MRD) after AML treatment in AML patients group. Results: In this study, 27 AML patients and 24 healthy individuals with mean age of 40.92±15.13 years were evaluated, of whom 56.86% were male. The mean bone marrow MVD was significantly higher in AML patients than controls. The mean bone marrow MVD was significantly higher in males and there was insignificant reverse correlation between bone marrow MVD and MRD. About 59.3% of AML patients had response to treatment and there was no significant relationship between MVD and response to treatment. Conclusion: Bone marrow MVD was higher in AML patients than controls and there was no remarkable relationship between bone marrow MVD and MRD and response to treatment.

Association of CD49d expression with clinicopathological features of chronic lymphocytic leukemia patients in the Iranian population.

BACKGROUND: Identification of factors affecting prognosis in chronic lymphocytic leukemia (CLL) is important for risk stratification of patients. METHODS: In the present study, CD49d expression was analyzed by multi-color flow cytometry in 98 newly diagnosed and untreated CLL patients at the hematopathology ward. The patients were divided into two subgroups according to CD49d expression (30% cut off) and the association of this marker with the patients' clinicopathological properties were evaluated. RESULTS: In this study, CD49d expression exhibited significant association with the Rai stage of the disease (P<0.0001), CD38 status (P<0.0001), hemoglobin level (P=0.0006), and platelet count (P=0.0016). The CD49d-positive patients presented in higher stages in comparison with CD49d-negative patients. Although only 1% of the CD49d-negative patients were CD38-positive, this proportion for CD49d-positive group was 69%. However, no significant correlation was observed between CD49d expression and patients' age (P=0.2031), gender (P=0.8119), and absolute lymphocytes count (P=0.1073). CONCLUSION: Therefore, CD49d is a grateful biomarker with high association with clinicopathological parameters in CLL patients.

Immune Checkpoint Molecules in Primary Diffuse Large B-Cell Lymphoma of the Central Nervous System.

INTRODUCTION: Primary Diffuse Large B Cell Lymphoma of CNS (PCNSL) is a rare variant of Diffuse Large B Cell Lymphoma (DLBCL) and presents with an aggressive clinical course and usually resistant to commonly used therapy regimens. Recently, role of immune checkpoint molecules including PD-1 and PD-L1 confirmed in some solid tumors and lymphoma resulting tumor cells escape the immune system and help to survive and to spread. Inhibitors of PD-1 and PD-L1 have shown lasting responses in several solid and some hematological tumors, while limited studies evaluate checkpoint molecules on PCNSL. METHOD: In this study, we investigated PD-1 and PD-L1 expression by immunostaining on 71 patients with PCNSL and correlation with demographic data, location of the tumor, proliferation rate, cell of origin, and CD8 positive T cell infiltration in tumor microenvironment. RESULTS: 16 from71 showed PD-1 expression, while PD-L1 expression were 42/71. No association was determined between PD-1/PD-L1 expression and gender, cell of origin, and proliferation rate, but a highly significant difference was determined between the infiltration of CD8 positive T cells in two groups of PD-1/PD-L1 positive and negative. CONCLUSION: This study revealed expression of check point molecules in remarkable number of PCNSL which may open new therapeutic recommendations in this aggressive lymphoma type.

Multiple Myeloma with Intracytoplasmic Azurophilic Granules.

Bone marrow examination plays an important role in the diagnosis of multiple myeloma. In some cases with multiple myeloma, marrow plasma cells with cytoplasmic inclusions are seen. In this study, a 46-year- old man was evaluated for multiple myeloma. In bone marrow aspiration, large intracytoplasmic azurophilic granules, resembling intracellular microorganisms were seen. IHC study demonstrated that these cells are CD138 positive. This is a rare histologic finding that usually results from the deposition of excess immunoglobulin.

The relationship between iron bone marrow stores and response to treatment in pediatric acute lymphoblastic leukemia.

Iron is an intracellular element whose accumulation in the body is associated with tissue damage. This study examines the effect of iron on pediatric acute lymphoblastic leukemia (ALL) and its "response to treatment." At the end of the first year of treatment, bone marrow iron store (BMIS) was evaluated in children with ALL and the relationship between iron store and minimal residual disease was investigated. Moreover, the 3-year disease-free survival (3-DFS) of patients was determined. Patients' BMIS were compared with that of subjects with normal bone marrow. The study examined 93 children, including 78 Pre-B and 15 T-cell ALL patients. BMIS did not differ between the children with ALL and those with no evidence of cancer. BMIS was increased in 26.6% of patients at the end of the first year of treatment. Drug resistance and BM relapses were more prevalent in cases with high BMIS in both Pre-B and T-cell groups. Bone marrow iron store is not considered a risk factor for childhood ALL. However, high levels of BMIS are associated with poor response to treatment and the risk of relapse. Bone marrow iron store control during treatment can therefore help achieve better outcomes and improve the chances of recovery.

Pseudohypercalcemia in Multiple Myeloma: A Case Report.

Hypercalcemia is a common finding in patients with multiple myeloma. Clinical manifestations of hypercalcemia correlate with the level of serum calcium. Ionized serum calcium (Ca (I)) will be increased in true hypercalcemia. In pseudohypercalcemia the Ionized Ca is normal, although binding of calcium to abnormal immunoglobulin causes increased serum calcium level. In the asymptomatic multiple myeloma patients with moderate to severe hypercalcemia, measurement of ionized calcium is critical to exclude pseudohypercalcemia. Here, we describe an asymptomatic 44-year-old man with multiple myeloma who had severe hypercalcemia, but normal serum Ionized Ca level.

The correlation of CD19 + CD24 + CD38 + B cells and other clinicopathological variables with the proportion of circulating Tregs in breast cancer patients.

BACKGROUND: T regulatory cells (Tregs) are known to negatively control immune response. The frequency of these cells was inversely correlated with clinical outcomes of breast cancer. CD19+CD24hiCD38hi cells also play a critical role in inflammation and autoimmune disease. However, their function in tumor immune response is less studied. In this study we aimed to determine the role of CD19+CD24hiCD38hi cells and some other clinicopathological variables in increasing the proportion of Tregs in breast cancer patients. METHODS: We selected 47 patients with invasive ductal breast carcinoma and 50 healthy controls and obtained their blood samples. RESULTS: The proportion of circulating CD4+CD25+Foxp3+ Tregs and CD19+CD24hiCD38hi cells was significantly increased in breast cancer patients. We also found that increased proportion of Tregs in breast cancer is correlated with HER2 amplification, advanced clinical stages, serum TGF-ß1 and increased CD19+CD24hiCD38hi cells in the peripheral blood. CONCLUSION: Altogether, our data suggest that as much as Tregs, CD19+CD24hiCD38hi B cells could also have a part in the suppression of immune response in breast cancer.