Reduction In Leukocyte Counts Research Articles

Adapting the protocol for studying the functional capacity of T lymphocytes thawed from cryopreservation

Background. Immunological studies are impossible without long-term storage of cryopreserved biomaterial. There are no standard procedures for working with cryopreserved mononuclear leukocytes.The aim of the study. To optimize the protocol for culturing T lymphocytes thawed after cryopreservation by assessing their viability and proliferative capacity.Methods. Mononuclear leukocytes were isolated from the peripheral blood of relatively healthy volunteers (n = 18). Cells were subjected to controlled freezing down to –80 °C and were transferred to liquid nitrogen. First step: after thawing, the cells were stained with CFSE (carboxyfluorescein succinimidyl ester), were divided into two parts and cultured in the presence/absence of interleukin 2 (IL-2). Cell proliferation was stimulated with phytohemagglutinin (type P). Cells were incubated for 7 days. Sample analysis was performed using flow cytometry. Second stage: thawed cells were divided into three parts. Two parts were resuspended in a full growth medium with IL-2 and were placed in a thermostat (+37 °C) to “rest” for one hour or overnight. After “resting”, the cells were stained with CFSE. One third of the thawed leukocytes were stained with CFSE immediately after thawing. Cells were stimulated, cultured and analyzed the same way at both stages of the study.Results. It has been established that adding IL-2 to the culture medium contributes to a better cell survival. In the presence of IL-2, stimulated CD4+ and CD8+ T lymphocytes produced more daughter cell generations. At the end of the 7-day incubation “rested” samples had reduced leukocyte counts compared to the samples that were cultured immediately after thawing. The number of daughter cell generations formed by stimulated CD4+ and CD8+ T cells decreased when the “rest” stage was included into the study protocol.Conclusion. Adding IL-2 into culture medium can increase the viability and mitotic capacity of thawed T cells, making their state more similar to that of freshly isolated lymphocytes. Cell “rest” after thawing negatively affects the viability and proliferative activity of T lymphocytes during their weekly incubation.

Andrographis paniculata Leaf Extract Increases Interleukin-2 in Malnutrition Rat Model

BACKGROUND: Malnutrition is a global health concern that results in changes in nutritional status, as indicated by alterations in phenotypic markers, hematological and biochemical parameters, and increased susceptibility to infection, as shown by decreased interleukin (IL)-2 levels. Andrographolide, the active component of Andrographis paniculata, stimulates the immune system and exhibits antibacterial and antiviral activity. Therefore, A. paniculata may serve as a potential adjuvant therapy for malnutrition. This study was conducted to analyze the effect of A. paniculata as an immunomodulator against malnutrition with characteristics of environmental enteric dysfunction (EED) and a low-protein diet by examining phenotypic markers, hematological, biochemical, and IL-2 levels.METHODS: Forty-five male Wistar rats were divided into seven groups. They were fed either a standard or a low-protein diet before receiving oral administration of various concentrations of A. paniculata leaf extract (APLE). APLE was administered 21 days after the initial low-protein diet. Hematological, biochemical, and phenotypic markers were assessed to determine the nutritional status of the rats. The protective effects of APLE were evaluated by measuring IL-2 levels using enzyme-linked immunosorbent assay (ELISA).RESULTS: Malnourished rats exhibited slow body growth, physical and behavioral changes, reduced leukocyte count, total protein, albumin, cholesterol, and villi length. Malnourished rats treated with APLE showed a more effective and significant increase in IL-2 levels, with higher concentrations of APLE resulting in higher IL-2 levels.CONCLUSION: APLE, in a concentration-dependent manner, can increase IL-2 levels, suggesting that APLE may have potential protective effects in a rat model of malnutrition.KEYWORDS: Andrographis paniculata, environmental enteric dysfunction, interleukin (IL)-2, low protein, malnutrition

Clinical Manifestations and Treatment Outcomes of Brucella Endocarditis: A Retrospective Cohort Study at a Tertiary Cardiac Centre in Bengaluru, Karnataka, India

Introduction: Brucella is a rare cause of Infective Endocarditis (IE), requiring prompt early recognition and treatment to prevent life-threatening complications. Diagnosis is often missed or delayed, leading to an increase in cardiac morbidity and mortality. Aim: To analyse the clinical profile, laboratory parameters, cardiac manifestations, management patterns, and outcomes of Brucella endocarditis. Materials and Methods: A retrospective cohort study was conducted at the Department of Cardiology, Sri Jayadeva Institute of Cardiovascular Sciences and Research, Bangaluru, Karnataka, India on patients with blood culture-confirmed Brucella endocarditis, diagnosed using modified Duke’s criteria between January 2010 and December 2021. The present study evaluated the clinical presentation, treatment modalities, and outcomes. Descriptive statistical analyses were performed. Results: A total of 34 cases were identified during the study period. The mean age of the patients was 35.6±14 years (age range: 15-66 years), with 82.4% males and 17.6% females. Underlying valvular heart disease was present in 82.4% of the patients, while 17.6% had no pre-existing valvular heart disease. Both aortic and mitral valves were involved with equal frequency. All patients presented with fever. Most patients had a normal leukocyte count (61.8%). Thrombocytopenia (32.3%) and pancytopenia (17.6%) were also observed. Large vegetations (>1 cm) were seen in 38.2% of patients, and complications related to IE were observed in 35.2%. The majority of patients (82.4%) were managed medically alone in the acute phase. The antibiotic regimen of doxycycline with rifampicin combined with intravenous gentamicin was used in the majority of the patients. The observed mortality rate was 17.6%. Conclusion: Brucella endocarditis can present with thrombocytopenia/pancytopenia along with normal or reduced leukocyte count. The addition of intravenous gentamicin to oral therapy may reduce relapse rates.

Therapeutic effects of medicinal plants and their constituents on lung cancer, in vitro, in vivo and clinical evidence.

The most common type of cancer in the world is lung cancer. Traditional treatments have an important role in cancer therapy. In the present review, the most recent findings on the effects of medicinal plants and their constituents or natural products (NP) in treating lung cancer are discussed. Empirical studies until the end of March 2022 were searched using the appropriate keywords through the databases PubMed, Science Direct and Scopus. The extracts and essential oils tested were all shown to effect lung cancer by several mechanisms including decreased tumour weight and volume, cell viability and modulation of cytokine. Some plant constituents increased expression of apoptotic proteins, the proportion of cells in the G2/M phase and subG0/G1 phase, and Cyt c levels. Also, natural products (NP) activate apoptotic pathways in lung cancer cell including p-JNK, Akt/mTOR, PI3/ AKT\ and Bax, Bcl2, but suppressed AXL phosphorylation. Plant-derived substances altered the cell morphology, reduced cell migration and metastasis, oxidative marker production, p-eIF2α and GRP78, IgG, IgM levels and reduced leukocyte counts, LDH, GGT, 5'NT and carcinoembryonic antigen (CEA). Therefore, medicinal plant extracts and their constituents could have promising therapeutic value for lung cancer, especially if used in combination with ordinary anti-cancer drugs.

Practical aspects of the use of ventoclax in combination with azacitidine for the treatment of newly diagnosed acute myeloid leukaemia in patients ineligible for intensive chemotherapy

The aim of treating elderly patients with acute myeloid leukaemia (AML) ineligible for intensive chemotherapy is to extend survival, but treatment results are often unsatisfactory. Therapy with venetoclax combined with azacitidine allowed remission in two-thirds of patients and significantly prolong the median overall survival. The treatment is increasingly used in clinical practice and establishes a new medical standard. The use of venetoclax in treating AML requires knowledge of drug use rules and their individualization. This review summarizes critical elements of the clinical practice of venetoclax use in combination with azacitidine regarding the dosing regimen, management of cytopenias during therapy and treatment adjustments to prevent drug-to-drug interactions. Treatment with venetoclax can cause the risk of tumor lysis syndrome (TLS), and therefore step-wise dose ramp-up is required with the prophylaxis of TLS and reduction in leucocyte count. Cytopenias that occur during the therapy affect most of the patients; nevertheless, it is not recommended to modify the treatment until the remission of the disease. In haematologic toxicity after disease remission, it is recommended to delay the next cycle and shorten the treatment while maintaining the dose. Knowledge about venetoclax drug-to-drug interactions is necessary for an efficacious and safe therapy. It is essential to reconsider the rationale behind using some agents, e.g., azole derivatives commonly used in the prophylaxis of invasive fungal infections, as well as to be aware of the rules of venetoclax dosing. Detailed knowledge of the above aspects of therapy is essential to ensure the continuity, safety, and efficacy of venetoclax with azacitidine.

Influence of Physical Exercise Program on Glycemic Status and Inflammatory Parameters in Type 2 Diabetes Mellitus

Introduction: Physical activity has a positive effect on the regulation of diabetes mellitus and has been shown to have benefits in improving health. Aim: Examine the effects of physical activity on changes in glycemic parameters (glucose, HbA1C) and inflammatory parameters (leukocyte count (WBC), CRP) before and one month after the exercise program. Material and Methods: The study was designed as a prospective, analytical, and observational study. Results: T-test of paired samples assessed the impact of physical activity on glycemic and inflammatory parameters in subjects with confirmed Diabetes mellitus type 2 and in subjects without confirmed diabetes mellitus type 2. In subjects with confirmed diabetes the value of eta squared (ɳ2 - eta sqared) is 95%, which indicates a significant impact of physical activity on the change in glucose values and on the change of HbA1C value was indicated by the value of eta square of 93%. When it comes to inflammatory parameters, the impact of physical activity was found in the reduced number of WBC (ɳ2 = 88%), and in CRP (ɳ2 = 90%). In subjects without confirmed diabetes mellitus, a significant effect of physical activity on the change in glucose (ɳ2 = 94%) and HbA1C (ɳ2 = 77%). The influence of physical activity on the reduction of leukocyte count was proven by ɳ2 - eta sqared test (ɳ2 = 66%), as well as a decrease in CRP (ɳ2 = 30%). Conclusion: This study showed a significant impact of physical activity on the reduction of elevated glycemic and inflammatory parameters.

A RANDOMIZED COMPARATIVE STUDY OF CONSERVATIVE VS PERCUTANEOUS NEEDLE ASPIRATION OF LIVER ABSCESS BASED ON PERCENTAGE OF ITS LIQUEFACTION.

Introduction:- Liver abscess is one of the common acute abdominal condition. Conservative management and percutaneous needle aspiration are the commonly used methods of management. But there is no common consensus between the physicians regarding best option out of these two. To Aim:- compare and to identify the better treatment modality among these two options for the treatment of liver abscess based on the percentage of liquefaction. A total of 140 patients with liver abscess were included in the study. T Patients and Methods:- hey were grouped in to Group A and B, based on percentage of liquefaction, they were further subdivided into conservative and aspiration subgroups and randomized. The outcomes of the two procedures were studied with respect to reduction in abscess cavity size, reduction in leukocyte count and patient's satisfaction. There was a signicant reduction in the ab Results:- scess size and leukocyte count in conservative subgroup of group-A, than conservative sub group of group B. Whereas there was signicant reduction abscess size and leucocyte count noted in aspiration sub group of Group B than aspiration sub group of Group A. Signicantly more number of the patients in conservative subgroup of group-A expressed their satisfaction, than conservative sub group of group B. Whereas signicantly more number of patients in aspiration sub group of the group B expressed their satisfaction than aspiration sub group of group A. present study concludes that conservative Conclusion:- treatment is also better option for liver abscess with 30-50% liquefaction, and aspiration is ideal for 50-70% liquefaction

Impact of Malarial Infection on Hematological Parameters in Patients at Tertiary Care Hospitals in D.I. Khan

Malaria is among the leading parasitic infections in world causing major health problems. Plasmodium, being a blood parasite, causes changes in blood parameters. This study was aimed at identifying the Plasmodium specie and measuring the changes that occurred in hematological parameters, in malaria patients. The sample size consisted of 109 individuals divided into two (Infected and Control) groups: 71 malaria-positive and 38 malaria-negative individuals. The diagnosis was based on staining and direct visualization of the parasite under a microscope. Blood CP was performed on the "Sysmex XP 300 analyzer". Non-normally distributed continuous variables were compared using the Mann-Whitney U test. An association between malaria and thrombocytopenia and anemia was noted but severe cases were not observed. Female patients were more Anaemic and the reduction in leukocyte count was also observed among malaria patients. Malaria causes changes in hematological parameters with thrombocytopenia during malarial infection. However, anemia was observed in female patients.

Impact of Abnormal Leukocyte Count in the Pathophysiology of Sickle Cell Anemia.

Sickle cell disease (SCD) is a hereditary disease that causes deoxygenated erythrocytes to stiffen, resulting in vaso-occlusive crises, endothelial damage, organ failure and systemic consequences. An abnormal leukocyte count is associated with a worse clinical picture in SCD. I studied the link between an abnormal leukocyte count and sickle cell disease. I used the PRISMA guidelines to conduct a systematic review of 5 different clinical literatures that discussed the link. Use of databases including Web of Science, Scopus, Cochrane Library, MEDLINE, ScienceDirect, EMBASE, and JSTOR to make our findings. SCD is a red cell disorder, but an elevated leukocyte count plays a significant role in its pathophysiology. Sickled red cells adhere better to leukocytes when compared to normal red cells, which explains the worse clinical picture seen in sickle cell patients. Hydroxyurea therapy inhibits the recruitment and invasion of neutrophils, which lowers vaso-occlusion. An elevated leukocyte count is responsible for worse clinical manifestations in SCD, eg, clinical stroke, acute chest syndrome, end organ damage, and infarction pain. Leukocytes attach to erythrocytes and vascular endothelium to cause vaso-occlusion, responsible for these effects. They also elaborate inflammatory mediators to aid the process. The higher the leukocyte count, the worse the effects. Hydroxyurea is very helpful in disrupting several pathways of leukocyte contribution to SCD pathophysiology. Therefore, more research on other pharmacologic agents that interfere with this disease process is necessary. Lastly, there is a need for further research into the effects of a reduced leukocyte count in SCD pathophysiology.

Hematotoxicity and Nephrotoxicity in Prostate Cancer Patients Undergoing Radioligand Therapy with [177Lu]Lu-PSMA I&T.

Simple SummaryRadioligand therapy (RLT) with prostate-specific membrane antigen (PSMA)-directed agents has shown remarkable results in patients with advanced prostate cancer. Our objective was to provide data on the side effect profile of PSMA-directed RLT using the therapeutic radiotracer [177Lu]Lu-PSMA I&T. We evaluated patients with castration-resistant metastatic prostate cancer treated with at least three cycles of [177Lu]Lu-PSMA I&T. A substantial fraction of the patients already had impaired renal function and/or reduced white blood cell counts at baseline, but the degree of nephrotoxicity or hematotoxicity under RLT was low. No severe toxicities occurred under RLT.(1) Background: Prostate-specific membrane antigen (PSMA)-directed radioligand therapy (RLT) has shown remarkable results in patients with advanced prostate cancer. We aimed to evaluate the toxicity profile of the PSMA ligand [177Lu]Lu-PSMA I&T. (2) Methods: 49 patients with metastatic, castration-resistant prostate cancer treated with at least three cycles of [177Lu]Lu-PSMA I&T were evaluated. Prior to and after RLT, we compared leukocytes, hemoglobin, platelet counts, and renal functional parameters (creatinine, eGFR, n = 49; [99mTc]-MAG3-derived tubular extraction rate (TER), n = 42). Adverse events were classified according to the Common Terminology Criteria for Adverse Events (CTCAE) v5.0 and KDIGO Society. To identify predictive factors, we used Spearman’s rank correlation coefficient. (3) Results: A substantial fraction of the patients already showed impaired renal function and reduced leukocyte counts at baseline. Under RLT, 11/49 (22%) patients presented with nephrotoxicity CTCAE I or II according to creatinine, but 33/49 (67%) according to eGFR. Only 5/42 (13%) showed reduced TER, defined as <70% of the age-adjusted mean normal values. Of all renal functional parameters, absolute changes of only 2% were recorded. CTCAE-based re-categorization was infrequent, with creatinine worsening from I to II in 2/49 (4.1%; GFR, 1/49 (2%)). Similar results were recorded for KDIGO (G2 to G3a, 1/49 (2%); G3a to G3b, 2/49 (4.1%)). After three cycles, follow-up eGFR correlated negatively with age (r = −0.40, p = 0.005) and the eGFR change with Gleason score (r = −0.35, p < 0.05) at baseline. Leukocytopenia CTCAE II occurred only in 1/49 (2%) (CTCAE I, 20/49 (41%)) and CTCAE I thrombocytopenia in 7/49 (14%), with an absolute decrease of 15.2% and 16.6% for leukocyte and platelet counts. Anemia CTCAE II occurred in 10/49 (20%) (CTCAE I, 36/49 (73%)) with a decrease in hemoglobin of 4.7%. (4) Conclusions: After PSMA-targeted therapy using [177Lu]Lu-PSMA I&T, no severe (CTCAE III/IV) toxicities occurred, thereby demonstrating that serious adverse renal or hematological events are unlikely to be a frequent phenomenon with this agent.

The effects of hydroxyurea on proinflammatory cytokine and tissue histopathology in an experimental sepsis model.

The diagnosis and treatment of sepsis are costly to healthcare services, and it is an important disease with high mortality rates. In the pathogenesis of sepsis, for which we still cannot provide a complete cure, there is increased cytokine release and organ damage. Hydroxyurea has been shown to reduce leukocyte counts, decrease inflammatory cytokines, and limit organ inflammation in ischemia-reperfusion models. This study aimed to evaluate leukocyte counts, interleukin-1 beta (IL-1β), IL-6, and tumor necrosis factor-alpha (TNF-α) cytokine values and organ inflammatory processes in hydroxyurea-treated rats with an experimental sepsis model. After ethical approval, rats were randomly divided into three groups, control (n= 7), sepsis (n= 7), and hydroxyurea (n= 7). Sepsis was created using the cecal ligation and puncture (CLP) method in rats other than in the control group. Rats in the hydroxyurea group received hydroxyurea (200 mg/kg) intragastrically, and the control and sepsis groups received sterile distilled water. IL-1β, IL-6, and TNF-α levels were measured at 0, 8, and 24 hours after CLP in all rats. Blood samples were collected at the time of sacrification 24 hours after CLP and analyzed for the complete blood count. Tissue specimens were taken for histopathologic examination. Cytokine levels (IL-1β, IL-6, TNF-α), white blood cell counts, and tissue damage were increased after the sepsis model in rats. It was found that the cytokine levels at the 8th hour, white blood cell count, and brain tissue damage in the hydroxyurea group were decreased significantly compared with the sepsis group. Early hydroxyurea treatment in rats with sepsis decreases proinflammatory cytokine (IL-1β, IL-6, and TNF-α) levels and thus reduces brain damage.

Possible Divergent Local and Peripheral Immunological Effects of Low-Dose Mesencure, an Enhanced Mesenchymal Cell Therapy, May Contribute to Its Success in Treating Severe Covid-19 Patients

Mesenchymal stromal cells (MSC) are widely investigated for treating ARDS in Covid-19. Nonetheless, these efforts are overshadowed by studies predating the pandemic that mostly failed to show MSC efficacy in ARDS and recent disappointments with repurposed MSC products. Relying on years of MSC-related experience, Bonus BioGroup developed MesenCure: An enhanced allogeneic MSC therapy for Covid-19, professionalized by a unique combination of culture conditions and optimized in ARDS-relevant models.MesenCure is currently evaluated in a Phase II study in severe Covid-19 patients and administered (IV) in three doses (1.5M cells/kg, d1, d3, d5). A Phase I/II study on ten severe patients demonstrated a significant improvement in ARDS-related parameters following MesenCure treatment. Patients were discharged within one day (median) following treatment, requiring no respiratory support. Speedy recovery from local inflammation was observed in these patients, demonstrated by a rapid reduction in diffuse lung pneumonia, from 55% of the lung area to 15% within 5-6 days from the first dose (p<0.01, Fig. A-C). A corresponding drop in CRP was detected (p<0.01), which returned to normal. A multivariate regression analysis revealed that the reduction in CRP was mainly associated with the number of doses administered and not and the time elapsed since the first dose.MesenCure efficacy may be attributable to the cells' de novo expression of the gene encoding for the IL-6 receptor, making them more responsive to inflammation than non-professionalized naïve MSC (NA-MSC); as well as >8-fold upregulation of the EDIL3 gene, encoding for an endogenous inhibitor of immune infiltration. A corresponding immunosuppressive effect of MesenCure MSCs was demonstrated in vitro, showing their ability to suppress T cells activation twice more effectively than NA-MSC. In this study, MesenCure inhibited the proliferation of primary CD4 T cells in a concentration-depended manner following non-specific activation. Over 98% inhibition was achieved in co-culture of 1:10 MSC-to-PBMC with an IC 50 of 6k MSC/200k PBMCs (r 2=1.00) compared to 12k NA-MSC/200k PBMCs (r 2=0.95). Comparable results were also obtained for CD8 T cells. Similarly, MesenCure inhibited ROS production by primary neutrophils remarkably fast and by up to 80% within less than 40 minutes following their activation (IC 50 = 19k MSC/200k neutrophils, r 2=1.00).In addition to local immunosuppressive outcomes, a significant increase in blood leukocytes was observed in patients treated with MesenCure (p<0.05, Fig. D-F). Further analysis suggested that the increase in total WBCs and neutrophils was associated with the number of MesenCure doses administered (p<0.05, Fig. G-H). In contrast, the increase in lymphocytes was time-dependent (R=0.72, Fig. I).The seemingly exclusively localized anti-inflammatory effects seen in severe patients treated with MesenCure were also observed in animal (murine) studies. An in vivo study in an acute lung injury model demonstrated a dose-dependent localized reduction in leukocyte counts in the lung fluids of animals treated with MesenCure (IV) using two dose levels. Relative to untreated animals, MesenCure reduced lung leukocyte counts by 35%-43% in animals treated with the low dose and by 62%-67% following high-dose MesenCure treatment (p<0.05). The leukocytes' clearance from the lungs was accompanied by a 41%-57% reduction in lung edema (p<0.05) following MesenCure treatment. Notably, NA-MSC did not achieve the same effect. Similar to our clinical findings, a significant increase was measured in neutrophil counts in animals treated with low-dose MesenCure (p<0.05), which decreased dramatically (p<0.01) in animals treated with a four-times higher dose.MesenCure is administered at a much lower dose compared to other MSC products administered at up to 10M cells/kg. Considering the increase in blood leukocytes measured in patients treated with low-dose MesenCure and comparable preclinical findings, our data suggest that low-dose MesenCure could elicit a potent local anti-inflammatory effect without suppressing, and even enhancing, peripheral immunity that is needed to fight the virus. Further research is inevitably required into the mechanism behind this phenomenon. However, our results indicate MesenCure's potential in relieving local inflammation while giving the patient a fighting chance against viremia. [Display omitted] DisclosuresBronshtein: Bonus BioGroup: Current Employment. Ben David: Bonus BioGroup: Current Employment. Novak: Bonus BioGroup: Current Employment. Kivity: Bonus BioGroup: Current Employment. Meretzki: Bonus BioGroup: Current Employment. Rozen: Bonus BioGroup: Consultancy.

Successful treatment of refractory cystitis associated with systemic lupus erythematosus with Belimumab

Lupus cystitis (LC) is a rare manifestation of SLE, the diagnosis of LC is challenging, especially in the absence of other systemic manifestations or the obvious disease activity index of SLE, further cystoscopy and bladder biopsy are crucial. The symptoms of cystitis could be controlled with high-dose GC, but in the process of GC reduction, the condition repeated. Here we report one case of refractory LC treated with Belimumab.The case is a 45-year-old female patient who had SLE and presented with urinary urgency, frequency and pain for more than 3 years. Laboratory assays revealed high ANA, reduced complement 3 level, proteinuria, significantly elevated Leukocyte esterase and leukocyte in urine, with the negative urinary culture of bacteria and mycoplasma, meanwhile, the cystoscopy and bladder biopsy showed interstitial inflammation. Confirming the diagnosis of refractory SLE-LC, recommended-dose Belimumab (10 mg/kg fortnightly for 3 times, monthly for 6 times) was administered, resulting in the normalization of urinary activity and significant reductions in leukocyte counts and protein levels of urine. No lupus cystitis relapse or SLE activity occurred during 10 months of follow-up.Our case confirmed the efficacy and good follow-up outcomes of Belimumab treatment for refractory SLE-LC.

Reduced SLIT2 is Associated with Increased Cell Proliferation and Arsenic Trioxide Resistance in Acute Promyelocytic Leukemia.

Simple SummaryIn solid tumors, the altered expression of embryonic genes such as the SLIT-ROBO family has been associated with poor prognosis, while little is known about their role in acute myeloid leukemia (AML). Previous studies reported frequent hypermethylation of SLIT2 mediated by the methyltransferase enzyme EZH2 and more recently the PML protein, which are commonly found to be aberrantly expressed in AML. Here, we aim to assess retrospectively the clinical relevance of the SLIT2 gene in acute promyelocytic leukemia, a homogenous subtype of AML. We demonstrated that reduced SLIT2 expression was associated with high leukocyte counts and reduced overall survival in different APL cohorts. STLI2 treatment decreased APL growth, while SLIT2 knockdown accelerated cell cycle progression and proliferation. Finally, reduced expression of SLIT2 in murine APL blasts resulted in fatal leukemia associated with increased leukocyte counts in vivo. These findings demonstrate that SLIT2 can be considered as a prognostic marker in APL, and a potential candidate for clinical studies of a more heterogeneous disease, such as AML.The SLIT-ROBO axis plays an important role in normal stem-cell biology, with possible repercussions on cancer stem cell emergence. Although the Promyelocytic Leukemia (PML) protein can regulate SLIT2 expression in the central nervous system, little is known about SLIT2 in acute promyelocytic leukemia. Hence, we aimed to investigate the levels of SLIT2 in acute promyelocytic leukemia (APL) and assess its biological activity in vitro and in vivo. Our analysis indicated that blasts with SLIT2high transcript levels were associated with cell cycle arrest, while SLIT2low APL blasts displayed a more stem-cell like phenotype. In a retrospective analysis using a cohort of patients treated with all-trans retinoic acid (ATRA) and anthracyclines, high SLIT2 expression was correlated with reduced leukocyte count (p = 0.024), and independently associated with improved overall survival (hazard ratio: 0.94; 95% confidence interval: 0.92–0.97; p < 0.001). Functionally, SLIT2-knockdown in primary APL blasts and cell lines led to increased cell proliferation and resistance to arsenic trioxide induced apoptosis. Finally, in vivo transplant of Slit2-silenced primary APL blasts promoted increased leukocyte count (p = 0.001) and decreased overall survival (p = 0.002) compared with the control. In summary, our data highlight the tumor suppressive function of SLIT2 in APL and its deteriorating effects on disease progression when downregulated.

Novel manifestations of immune dysregulation and granule defects in gray platelet syndrome

AbstractGray platelet syndrome (GPS) is a rare recessive disorder caused by biallelic variants in NBEAL2 and characterized by bleeding symptoms, the absence of platelet α-granules, splenomegaly, and bone marrow (BM) fibrosis. Due to the rarity of GPS, it has been difficult to fully understand the pathogenic processes that lead to these clinical sequelae. To discern the spectrum of pathologic features, we performed a detailed clinical genotypic and phenotypic study of 47 patients with GPS and identified 32 new etiologic variants in NBEAL2. The GPS patient cohort exhibited known phenotypes, including macrothrombocytopenia, BM fibrosis, megakaryocyte emperipolesis of neutrophils, splenomegaly, and elevated serum vitamin B12 levels. Novel clinical phenotypes were also observed, including reduced leukocyte counts and increased presence of autoimmune disease and positive autoantibodies. There were widespread differences in the transcriptome and proteome of GPS platelets, neutrophils, monocytes, and CD4 lymphocytes. Proteins less abundant in these cells were enriched for constituents of granules, supporting a role for Nbeal2 in the function of these organelles across a wide range of blood cells. Proteomic analysis of GPS plasma showed increased levels of proteins associated with inflammation and immune response. One-quarter of plasma proteins increased in GPS are known to be synthesized outside of hematopoietic cells, predominantly in the liver. In summary, our data show that, in addition to the well-described platelet defects in GPS, there are immune defects. The abnormal immune cells may be the drivers of systemic abnormalities such as autoimmune disease.

Intake of snacks containing curcumin stimulates erythropoiesis and antioxidant response in dogs

The objective of this study was to determine whether snacks containing curcumin have beneficial effects on dog health. The snacks were produced from commercial canned meat for dogs, and curcumin was added and homogenized. Snacks containing 15 mg of curcumin were produced, frozen, and offered to dogs twice a day. Ten beagles (6 months of age) were used. The animals were placed in an experimental kennel and allocated randomly to 1 of 2 treatments (5 dogs/treatment): snacks containing curcumin (curcumin; 30 mg of curcumin/animal/day) or not (control). On day 15, numbers of erythrocytes and hematocrit were greater in dogs fed with curcumin than in control dogs. Dogs fed with curcumin had lower numbers of leukocytes (day 30), neutrophils (day 15), and lymphocytes (day 30) than did control dogs. Dogs fed with curcumin had lower plasma levels of nitric oxide, reactive oxygen species, lipoperoxidation, and protein carbonylation on day 30 than did control dogs. Finally, dogs fed with curcumin had greater plasma total antioxidant capacity and concentrations of protein thiol, non-protein thiol, glutathione peroxidase, and superoxide dismutase on day 30 compared with control dogs. We conclude that curcumin in dog snacks stimulated the antioxidant system and consequently reduced oxidative reactions, which is beneficial to animal health. Furthermore, 30 mg of curcumin/dog/day reduced leukocyte counts, which suggests mild anti-inflammatory effects.

Oral treatment with the Chinese herbal supplements B307 enhances muscle endurance of ICR mice after exhaustive swimming via suppressing fatigue, oxidative stress, and inflammation.

Exhaustive exercise may damage muscles due to oxidative stress and inflammation and cause muscle fatigue and soreness. The study investigated the effects of Chinese herbal supplements (CHS) B307 on muscle endurance after exhaustive swimming (ES). Thirty‐two male ICR mice were randomly divided into 4 groups: Sham + ES, pretreatment of CHS B307 + ES (Pre + ES), post‐treatment of CHS B307 + ES (Post + ES), and dual treatment of CHS B307 + ES (Dual + ES). All mice were subjected to ES in the form of a forced swimming test. Then, we compared ES time (EST) as the index of muscular endurance. Also, we examined the fatigue, oxidative stress, inflammation, and damage in the muscle tissue among these groups by using immunohistochemistry (IHC), chemiluminescence, and biochemical analysis. Our results revealed that those mice of Pre + ES and Dual + ES groups had remarkably better EST than those mice of Sham + ES and Post + ES groups. Those mice with oral treatment of CHS B307(Pre + ES, Post + ES, and Dual + ES groups) showed significantly reduced leukocyte counts in the urine, and reduced levels of reactive oxygen species (ROS), neutrophils, and lactic acid in the blood than those mice of Sham + ES. In addition, those mice with oral treatment of CHS B307 (Pre + ES, Post + ES, and Dual + ES groups) showed significant alleviation of oxidative stress, inflammation, and damage in the muscle tissue than those mice of Sham + ES. Thus, we suggested that CHS B307 can be a functional sports supplement because it can enhance muscle endurance after exhaustive swimming via suppressing fatigue, oxidative stress, and inflammation.

Melanocortin 3 receptor activation with [D-Trp8]-γ-MSH suppresses inflammation in apolipoprotein E deficient mice

The melanocortin MC1 and MC3 receptors elicit anti-inflammatory actions in leukocytes and activation of these receptors has been shown to alleviate arterial inflammation in experimental atherosclerosis. Thus, we aimed to investigate whether selective targeting of melanocortin MC3 receptor protects against atherosclerosis. Apolipoprotein E deficient (ApoE−/−) mice were fed high-fat diet for 12 weeks and randomly assigned to receive either vehicle (n = 11) or the selective melanocortin MC3 receptor agonist [D-Trp(8)]-gamma-melanocyte-stimulating hormone ([D-Trp8]-γ-MSH; 15 μg/day, n = 10) for the last 4 weeks. Lesion size as well as macrophage and collagen content in the aortic root plaques were determined. Furthermore, leukocyte counts in the blood and aorta and cytokine mRNA expression levels in the spleen, liver and aorta were quantified. No effect was observed in the body weight development or plasma cholesterol level between the two treatment groups. However, [D-Trp8]-γ-MSH treatment significantly reduced plasma levels of chemokine (C–C motif) ligands 2, 4 and 5. Likewise, cytokine and adhesion molecule expression levels were reduced in the spleen and liver of γ-MSH-treated mice, but not substantially in the aorta. In line with these findings, [D-Trp8]-γ-MSH treatment reduced leukocyte counts in the blood and aorta. Despite reduced inflammation, [D-Trp8]-γ-MSH did not change lesion size, macrophage content or collagen deposition of aortic root plaques. In conclusion, the findings indicate that selective activation of melanocortin MC3 receptor by [D-Trp8]-γ-MSH suppresses systemic and local inflammation and thereby also limits leukocyte accumulation in the aorta. However, the treatment was ineffective in reducing atherosclerotic plaque size.

Clinical and computed tomographic imaging features of novel coronavirus pneumonia caused by SARS-CoV-2

SummaryPurposeTo investigate the clinical and imaging characteristics of computed tomography (CT) in novel coronavirus pneumonia (NCP) caused by SARS-CoV-2.Materials and methodsA retrospective analysis was performed on the imaging findings of patients confirmed with COVID-19 pneumonia who had chest CT scanning and treatment after disease onset. The clinical and imaging data were analyzed.ResultsFifty patients were enrolled, including mild type in nine, common in 28, severe in 10 and critically severe in the rest three. Mild patients (29 years) were significantly (P<0.03) younger than either common (44.5 years) or severe (54.7) and critically severe (65.7 years) patients, and common patients were also significantly (P<0.03) younger than severe and critically severe patients. Mild patients had low to moderate fever (<39.1 °C), 49 (98%) patients had normal or slightly reduced leukocyte count, 14 (28%) had decreased counts of lymphocytes, and 26 (52%) patients had increased C-reactive protein. Nine mild patients were negative in CT imaging. For all the other types of NCP, the lesion was in the right upper lobe in 30 cases, right middle lobe in 22, right lower lobe in 39, left upper lobe in 33 and left lower lobe in 36. The lesion was primarily located in the peripheral area under the pleura with possible extension towards the pulmonary hilum. Symmetrical lesions were seen in 26 cases and asymmetrical in 15. The density of lesion was mostly uneven with ground glass opacity as the primary presentation accompanied by partial consolidation and fibrosis.ConclusionCT imaging presentations of NCP are mostly patchy ground glass opacities in the peripheral areas under the pleura with partial consolidation which will be absorbed with formation of fibrotic stripes if improved. CT scanning provides important bases for early diagnosis and treatment of NCP.

Comparison of the role of ultrasound guided transhepatic aspiration of gallbladder versus conservative management of acute calculous cholecystitis

Background: Gall stone disease remains one of the most common medical problem leading to surgical intervention. Cholecystitis accounts for 3-10% of abdominal pain worldwide. Acute cholecystitis is the most common complication of cholelithiasis accounting for 14 to 30% of cholecystectomies performed in many countries. Symptoms in cholecystitis are due to impaction of stone and subsequent distention of gallbladder with inflammation. Study is aimed to clarify the role of ultrasound guided transhepatic gallbladder aspiration in the early management of acute calculous cholecystitis.Methods: The study was conducted in total of 40 patients presenting with acute cholecystitis. 20 patients underwent ultrasound guided transhepatic aspiration of gallbladder with antibiotics (group A) and 20 patients were given antibiotics only (group B). Data were collected before intervention and post intervention duration of stay, pain according to visual analog scale, leucocytosis and fever were recorded for analysis. No complications were related to aspiration procedure.Results: Both groups were comparable. Group A patients had better pain relief (p=0.0001 day on 2 and p=0.004 on day 3 post aspiration), percentage reduction of leucocyte count (p=0.041 on day 3) and duration of hospital stay (p=0.004) which were statistically significant.Conclusions: Ultrasound guided transhepatic aspiration of gall bladder with antibiotics in acute cholecystitis results in better pain profile, faster reduction in leucocyte count and shorter duration of hospital stay when compared to antibiotics alone.