Platelet Research Articles

LABORATORY PREDICTORS FOR DIAGNOSING COLORECTAL ANASTOMOTIC LEAKAGE.

An important concern in oncological coloproctology is colorectal anastomotic leakage (AL), which occurs in 3.5%-21% of patients. Predicting the occurrence of failure based on the results of laboratory markers can be decisive for the treatment of this complication. To improve the early diagnosis of AL by establishing combinations and threshold values of laboratory markers - predictors of the inflammatory process. The prospective study, conducted from 2020 to 2023, included 213 rectal cancer patients who underwent low anterior resection after neoadjuvant chemoradiotherapy. The inflammatory biomarkers were assessed before surgery and on the 3rd, 5th, and 7th days of the postoperative period. AL diagnosed in 25 (11.74%) patients by the grade of severity was as follows: A (radiological) in 7 (3.29%) patients; B (clinical) - 4 (1.88%); C (clinically expressed, peritonitis) - 11 (5.16%), and P (late) - 3 (1.41%) patients. The changes in the laboratory indicators of the inflammatory response such as С-reactive protein (CRP), procalcitonin (PCT), the counts of neutrophils (NEU), lymphocytes (LYM), platelets (PLT), and neutrophil/lymphocyte ratio (NLR) were significant only in B or C AL grades. Among them, only three indicators were identified as significant for predicting AL when assessed 24 h before the onset of this complication, namely LYM (threshold value ≤ 0.97 × 103/mm3, sensitivity 66.7% and specificity 81.3%, p < 0.001); PLT (threshold value > > 257 103/mm3, sensitivity 58.6%, and specificity 86.7%, p < 0.001); and NLR (threshold value > 4.42, sensitivity 58.1%, and specificity 86.7%, p < 0.001). The three-factor model based on these selected indicators was set up, and the prognosis index (Prog) was proposed with the decision threshold Progcrit = 2.23. The sensitivity of the model was 80% (95% CI 51.9%-95.7%), and the specificity - 74.2% (67.6%-80.2%). Based on the routine laboratory predictors used in the complex diagnosis of AL, B or C AL grades may be predicted allowing for the timely effective early diagnosis, medication, and surgical intervention..

EFFICACY OF CHROMOLAENA ODORATA SUPPLEMENTATION ON METHOTREXATE -INDUCED NEPHROPATHY AND OXIDATIVE STRESS

We carried out this research to investigate nephro-protective efficacy of Chromolaena odorata aqueous leaf extract on kidney toxicity in Methotrexate (MTX)-induced albino male rats. Thirty (30) wistar male rats spread into five study groups were used. Group I: orally administered normal saline daily and was the normal control. Group II: received C. odorata extract (250 mg/kg B.W.) orally once daily for ten days. Group III: received MTX (7mg/kg B.W.) intraperitoneally from 8th day for three days consecutively. Group IV: received C. odorata extract (250 mg/kg B.W.) orally once daily for ten days and then administered MTX (7 mg/kg B.W.) intraperitoneally from 8th day for three days consecutively. Group V: received Vitamin C (100 mg/kg B.W.) orally once daily for ten days and then administered MTX (7 mg/kg BW) intraperitoneally from 8th day for three days consecutively. 24hours after last treatment and administration, all rats were sacrificed and blood collected in EDTA bottles for hematology or in plain tubes for biochemical analysis. Creatinine and Urea were analyzed in serum. Superoxide Dismutase (SOD), Catalase, Malondialdehyde (MDA) and histopathological analysis was assessed using the kidney tissues. The obtained results revealed that three days of consecutive Methotrexate administration caused a significant rise in kidney function activity as well as severe oxidative damage and oxidative stress shown by a significant rise in Creatinine and Urea, increased level of MDA and a significant drop in SOD and CAT. However, C. odorata attenuated Methotrexate-induced biochemical alterations and oxidative stress in kidney. Also, C. odorata alone or in combination with Methotrexate showed an increase in Packed Cell Volume (PCV), Red Blood Cells (RBC), Haemoglobin (HB), White Blood Cells (WBC) and Platelets level compared to the group administered Methotrexate alone which showed degenerative levels in the analyzed hematological parameters. Showing corroboration, histopathological analysis showed deterioration and collapse of the kidney tubules in rats administered Methotrexate alone whereas C. odorata groups showed considerable nephro-protection and restoration towards normalcy. The results from this study revealed that the C. odorata have a nephro-protective efficacy against Methotrexate-induced kidney toxicity and damage via scavenging of free radicals and stabilization/enhancement of endogenous antioxidant status

Fibromiyalji Sendromunda Sistemik İmmün-İnflamasyon İndeksi ve Hematolojik Laboratuvar Bulgularına Genel Bakış

Bu çalışmada fibromiyalji sendromunda (FMS) Sistemik İmmün-İnflamasyon İndeksi (SII) ile kan parametreleri arasındaki ilişki değerlendirilerek hastalık aktivitesinin belirlenmesi amaçlanmıştır. Çalışmaya Ocak 2021 ve Ocak 2022 tarihleri arasında Fizik Tedavi ve Rehabilitasyon Kliniğine başvuran 109 FMS hastası ve 82 sağlıklı kontrol dahil edilmiştir. Yaş, cinsiyet, C-reaktif protein (CRP), eritrosit sedimentasyon hızı (ESR), beyaz kan hücresi (WBC), hemoglobin (HGB), ortalama korpusküler hacim (MCV), ortalama korpusküler hemoglobin (MCH), ortalama korpusküler hemoglobin konsantrasyonu (MCHC), kırmızı hücre dağılımı (RDW), trombosit dağılım genişliği (PDW), ortalama trombosit hacmi (MPV), trombosit (PLT), trombosit kriteri (PCT), lenfosit (LYM), monosit (MONO), nötrofil (NEU) düzeyleri hastane bilgi sisteminden retrospektif olarak taranmıştır. Trombosit-lenfosit oranı (PLR), nötrofil-lenfosit oranı (NLR), monosit-lenfosit oranı (MLR) ve sistemik immün-inflamasyon indeksi (SII) hesaplanmıştır. CRP ve ESR düzeyleri FMS hastalarında sağlıklı kontrollere göre daha yüksekti ancak değer aralığının dışında değildi (p&amp;lt;0,001). PDW (p&amp;lt;0,001), HGB (p&amp;lt;0,001), MCV (p&amp;lt;0,001), MCH (p&amp;lt;0,001) ve MCHC (p=0,02) düzeyleri hastalarda sağlıklı gruba göre daha düşüktü. Hasta ve kontrol grupları arasında yaş, WBC, NEU, LYM, MONO, RDW, MPV, PLT, PCT, SII, PLR, NLR ve MLR değerleri açısından istatistiksel olarak anlamlı fark yoktu. FMS hastalarında WBC düzeyleri ile SSS değerleri arasında (r=0,2; p=0,005) ve lenfosit düzeyleri ile WPI değerleri arasında (r=0,2; p=0,01) anlamlı pozitif korelasyon bulunmuştur. Çalışma sonuçlarına göre, SII'nin FMS'de belirleyici bir rolü yoktur, ancak hastalığın tanıda yararlı olabilecek bazı inflamatuvar bileşenleri vardır ve daha ayrıntılı çalışmalara ihtiyaç vardır.

A novel approach to simultaneous genotyping of human platelet antigen systems and human leucocyte antigen class I loci using PacBio long-read sequencing.

Accurate human leucocyte antigen (HLA) and human platelet antigen (HPA) typing is essential for establishing a blood platelet donor bank to deal with refractoriness in patients undergoing multiple platelet transfusions. Current methods, such as Sanger and next-generation sequencing, encounter difficulties in haplotyping. Herein, the aim of this study was to establish a method for HLA and HPA typing based on the long read sequencing. The HPA and HLA class I genotypes of 268 platelet donors from the Taiyuan Blood Center, China were identified using long-read sequencing on the PacBio platform. Allele frequencies for HPA systems and HLA class I genes were calculated, and genetic variability within HPA system genes was analysed. Polymorphisms were identified in 8 of the 35 HPA systems (HPA-1 to HPA-6w, HPA-15 and HPA-21w), with the frequencies of the 'b' allele at 0.0187, 0.0709, 0.4086, 0.0075, 0.0149, 0.0317, 0.4310 and 0.0019, respectively. The alleles with the highest frequencies at the HLA-A, HLA-B and HLA-C loci are HLA-A02:01, B51:01, B46:01 and C06:02, respectively. Additionally, several genetic patterns in HPA systems were identified, including the c.166-1029C>T variant, which was found exclusively in samples carrying the HPA-1b allele. This study developed a targeted long-read sequencing method characterized by high throughput and simultaneity, capable of resolving allele ambiguities for effective HLA class I genotyping in establishing a platelet donor bank.

The importance of genetic variant cleaners: From patient to wet lab and back to clinical practice.

Laureano J. Kamiya and colleagues have identified two novel missense variants (Gly168Arg and Gly168Glu) in the RUNX1 gene. These variants, identified in two unrelated families with familial platelet disorder with a predisposition to acute myeloid leukaemia (FPD/AML) phenotype, were initially classified as variants of uncertain significance (VUS). However, functional studies of RUNX1 target genes enabled their reclassification as likely pathogenic. The findings highlight Gly168 as a new mutation hotspot in RUNX1, providing important insights for genetic counselling and leukaemia monitoring. The study emphasizes the necessity for continuous updates to diagnostic guidelines and data sharing among laboratories to improve the classification and interpretation of genetic variants. Commentary on: Kamiya etal. Two novel families with RUNX1 variants indicate glycine 168 as a new mutational hotspot: Implications for FPD/AML diagnosis. Br J Haematol 2024 (Online ahead of print). doi: 10.1111/bjh.19776.

The importance of blood parameters in the detection of intestinal metaplasia and early diagnosis of gastric cancer

Background: We aimed to determine whether complete blood count(CBC) parameters, such as the white blood count(WBC), hemoglobin(Hb), platelet(PLT), red cell distribution width(RDW), mean platelet volume(MPV), platelet distribution width(PDW), neutrophil-to-lymphocyte ratio(NLR), platelet-to-lymphocyte ratio(PLR), and monocyte-to-lymphocyte ratio(MLR), have a predictive value in the detection of gastric cancer(GC) and intestinal metaplasia(IM). Methods: While proven GC, IM, and healthy control(HC) patients were included, patients with the comorbid disease were excluded, and univariate analyses compared three groups. The receiver operating characteristic(ROC) curve analysis was run for CBC parameters. The area under the curve(AUC) was evaluated, and a cut-off value was determined. The sensitivity and specificity of each parameter were considered. Results: The GC, IM, and HC groups consisted of 72(33%), 73(34%), and 72(33%) patients, respectively. RDW, PLT, NLR, PLR, and MLR were significant between GC and IM. The highest AUC (0.727) was obtained for the PLT yielding a 56.9% sensitivity and 79.4% specificity at a cut-off value of 151.8. The AUC of RDW was found as 0.691 and 0.626 for pairwise comparisons of GC-HC and IM-HC, respectively. At a cut-off value of 13.4, PLR yielded 70.8% sensitivity and 61.1% specificity in the detection of GC, while 64.4% sensitivity and 51.1% specificity for IM. Conclusion: CBC parameters, such as RDW, PLT, NLR, PLR, and MLR, have value in detecting GC. RDW also has diagnostic value in helping to detect IM. PLR can help to distinguish patients with GC from those with IM. These inexpensive, easily accessible parameters may help in the timely diagnosis of GC and IM. Keywords: Gastric cancer; intestinal Metaplasia; Monocyte to lymphocyte ratio; red cell distribution; hemoglobin; platelet; neutrophil-lymphocyte ratio; platelet-lymphocyte ratio; inflammation.

Umbilical Cord Blood Platelet Lysate Eyedrops for the Treatment of Severe Ocular Surface Disorders in Graft vs. Host Disease Patients: Clinical Study

Background: Graft-versus-host disease (GvHD) is an overactive systemic inflammatory response that can arise following allogeneic hematopoietic stem cell transplantation (HSCT). This condition occurs when the transplanted donor immune cells recognize the recipient’s tissues as foreign and trigger an immune response against them. The ocular surface (eyelids, conjunctiva, meibomian glands, lacrimal glands, and cornea) is particularly involved in GvHD, and its response to existing treatments, including potent immunosuppressants and new targeted therapies, is undesirable, with such treatments often being ineffective. Human allogeneic umbilical cord blood platelet lysate stands out as a potent adjunct to conventional therapies for ocular surface disorders related to severe Dry Eye Disease. This study aimed to evaluate the safety and efficacy of umbilical cord blood platelet lysate eyedrops for the treatment of severe ocular surface disorders in graft-versus-host disease patients who have received previous unsuccessful treatments. Methods: This study was a prospective, non-comparative, interventional case series study involving 22 patients (10 females and 12 males) aged 25–46 years with severe ocular surface disorders that were unresponsive to standard treatments. The GvHD patients were categorized based on the severity of their ocular surface disorders into three groups: Group I: five patients with severe Dry Eye Disease and filamentary keratitis; Group II: eight patients suffering from severe blepharo-kerato-epitheliopathy; Group III: nine patients with corneal ulcers. Fresh umbilical cord blood (UCB) was obtained from healthy donors and subjected to centrifugation using a novel PRP preparation kit provided by Sciacca (AG) Cord blood bank, Italy in a one-step process. In all groups, the outcomes before and after treatment were evaluated by means of the OSDI (Ocular Surface Disease Index), SANDE (Symptom Assessment in Dry Eye) questionnaire, VAS (Visual Analogue Scale), slit lamp examination, Esthesiometry, Lissamine Green Staining, the NIBUT (Non-Invasive Break-Up Time) and BUT, fluorescein staining with digital photography and Oxford classification, the Schirmer Test, the Best Corrected Visual Acuity (BCVA), and Meibography. In Group III at each evaluation time, the size of the ulcer and its relative reduction compared to the baseline size were recorded. Clinical variables, such as corneal inflammation, conjunctivalization, corneal neovascularization, or pain, were also considered individually. Results: We observed a significant improvement in the SANDE, VAS, and OSDI scores; Schirmer Test; BUT; BCVA; and Oxford classification after treatment with allogeneic cord blood serum eyedrops. Nevertheless, pain and inflammation reduced markedly over time until complete healing in all cases. The mean reduction in the ulcer surface area (compared to baseline values) was significantly higher at all assessment points (p = 0.001 for day 7 and p &lt; 0.001 for subsequent time points every 30 days for 90 days). At the last check-up (after 90 days of treatment), the number of ulcers (Group III, nine patients) with a reduction in size of greater than 50% was eight (88.8%), of which seven ulcers were completely healed. None of the patients experienced treatment-related local or systemic adverse events. In this study, using a relatively large number of cases, we demonstrated that the use of umbilical cord blood platelet lysate eyedrops is a safe, feasible, and effective curative approach for severe ocular surface disease in patients with GvHD. Conclusions: Our pilot study highlights the remarkable effectiveness of allogeneic cord blood serum eyedrops in patients with severe ocular surface disorders following GvHD who have shown an inadequate response to the usual treatments. It is mandatory to design future studies on the efficacy of this therapeutic approach for acute ocular, mucosal, and cutaneous GvHD.

Methanolic stem-bark extracts of Adansonia digitata modulates haematological and antioxidant parameters in streptozotocin-induced diabetic rats

Background and AimDiabetes mellitus is associated with haematological abnormalities and oxidative stress, contributing to disease progression and complications. Adansonia digitata, is a traditional medicinal plant well known for its potential therapeutic properties. This study investigates the effects of methanolic stem-bark extracts of Adansonia digitata on haematological and antioxidant status in streptozotocin-induced diabetic rats. Experimental ProcedureFourty-two wistar rats were distributed into 6 groups of 7 each. Group 1 served as normal control, group 2 to 6 animals were made diabetic by intraperitoneal administration of single dose of 50 mg kg-1 streptozotocin. Group 2 served as the negative control and did not receive treatment, group 3 served as the positive control and received 150 mg kg-1 metformin (Glucophage) (standard drug), while group 4, 5 and 6 received oral treatment of 100 mg kg-1, 200 mg kg-1 and 300 mg kg-1 of Adansonia digitata stem-bark methanolic extract respectively for 21days, after which the animals were sacrificed, and blood and liver were collected for haematological analysis including White Blood cell (WBC), red blood cell (RBC), haemoglobin (HGB), hematocrit (H CT), platelet (PLT), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), total bilirubin (TB), direct bilirubin(DB) and indirect bilirubin (IB) concentration and antioxidant parameters including antioxidant enzymes [catalase (CAT) and superoxide dismutase (SOD) activities] and oxidative stress marker [thiobarbituric acid reactive substance (TBARS)]. ResultsSTZ administration evoked a marked (p < 0.05) decline in the level of WBC, RBC, HGB, PLT, LYM and MCV in the diabetic rats when compared with the normal control. Howbeit, treatment with methanolic extracts of adansonia digitata stem-bark led to a profound (p < 0.05) increase in concentrations of these parameters close to the normal. In addition, STZ caused a non-significant (p > 0.05) decrease in MCH and MCHC. On the contrary, STZ caused an insignificant (p > 0.05) increase in the levels of TB and DB, but a significant (p > 0.05) increase IDB level in the diabetic control group when compared with the normal control group, while treatment with stem-bark extracts resulted in a marked (p > 0.05) fall in IDB level. Furthermore, there was a marked (P < 0.05) reduction in the activity of hepatic CAT and SOD and a considerable (P < 0.05) elevation in Malondialdehyde (MDA) level in the hepatic tissue of the diabetic control. Nonetheless, Adansonia digitata stem-bark extracts profoundly (P < 0.05) raised the activities of the enzymes and concomitantly counteracted MDA level. The extract elicited strong potency at 300 mg kg-1 which is comparable to metformin (Glucophage). ConclusionConsequently, this study advocates for the utility of Adansonia digitata extracts for the development of novel drugs for combating diabetes mellitus, associated hematological and oxidative stress complications and other ailments. Further research is needed to evaluate the clinical relevance of these effects in human subjects and to provide information on the sustained efficacy of the extracts in chronic models. Significant statementOxidative stress and hematological abnormalities are two of the consequences linked to diabetes mellitus. Anaemia and poor blood coagulation are two haematological changes that increase the risk of diabetes-related illness and death. Furthermore, oxidative stress is essential to the pathophysiology of diabetic complications such as nephropathy and cardiovascular disease. In addition, there are reports on hematological and biochemical alterations in diabetic subjects arising from the intake of synthetic antidiabetic medications.. Consequent to the safety, ease of use, efficacy, and lack of cytotoxic side effects of plants’ herbs, the study of Adansonia digitata's methanolic stem-bark extracts on haematological and antioxidant parameters in streptozotocin-induced diabetic rats is of paramount importance as the extracts' capacity to modify these parameters, will lead to the development of novel drugs that cures anemia, other blood abnormalities, and oxidative stress linked to diabetes as well as other ailments.

Growth performance, blood health, and antioxidant status of freshwater carp under brackish water rearing system: Sustainable aquaculture approach in arid and semi-arid conditions

Global warming, water salinization, and increased anthropogenic usage of freshwater sources have had severe effects on water sources and aquaculture practices, which is consequently putting pressure on aquatic fauna and fish production. Hence, brackish water can be used as an alternative and sustainable approach to meet the increasing food requirements. Four groups with triplicates were designed to evaluate the impact of brackish water on Cirrhinus mrigala (n = 100) for 180 days; T0: control group (freshwater), T1: (5.45 ppt salinity), T2: (8.45 ppt salinity) and T3 (12.45 ppt salinity). Little behavioural alterations and normal feeding were observed in all experimental groups as compared to control group, except group T3. Survival rate was 100 % in all groups excluding group T3, which had shown 88 % survival rate (P < 0.05), while mean weight gain and length gain were observed better with low FCR in T1 and T2 groups (P < 0.05) but best growth rate observed at 5 ppt salinity level. Significant reduction in haemoglobin (Hb), hematocrit (HCT), red blood cells (RBCs) and increase in white blood cells (WBCs) and platelets (PLTs) was observed in group T3 compared to other experimental groups (P < 0.05). ALT, AST, ALP, urea, creatinine, and triglycerides, exhibited significant changes in T3 group after 180 days (P < 0.05). Erythrocytes had shown morphological alterations as; micronuclei, notching, blebbing, pear-shaped nuclei and spherocytes in T3 group. Significant increase in DNA damage in various tissues had indicated potential genotoxic impact of highly saline brackish water. Oxidative stress markers were increased significantly and antioxidants reduced (P < 0.05) at high salinity level brackish water (T3) until 180 days. In conclusion, C. mrigala can potentially grow well in brackish water system at 5 ppt salinity whereas, rise in salinity level (12 ppt) is detrimental to freshwater fish health and growth.

A rare occurrence of Vancomycin-induced gastrointestinal hemorrhage without thrombocytopenia: a case report and literature review

BackgroundVancomycin-induced bleeding has been reported, attributed to the mechanism of immune thrombocytopenia. A rare case of vancomycin-induced gastrointestinal hemorrhage in a young patient with no underlying disease, receiving intravenous vancomycin for methicillin-resistant Staphylococcus aureus (MRSA) infection, is presented. This occurrence occurred without thrombocytopenia. Relevant cases reported in the literature were also reviewed.Case presentationA 34-year-old male patient presented with maxillofacial multiple spaces infection accompanying left temporal abscess, bilateral lung abscesses. Culture results from both blood and secretion indicated that the infection was caused by MRSA. The patient received standard-dose vancomycin (1 g q12h intravenously guttae) for treatment. On the 5th day of therapy, he presented with bright red blood in his stool; however, vancomycin treatment was continued. By the 9th day, a decrease in hemoglobin level to 76 g/L and a platelet (PLT) count of 424 × 109/L raised concerns about gastrointestinal hemorrhage. The hemoglobin level decreased to 62 g/L on day 12. Due to the high tissue concentration of linezolid, administration of linezolid at a dose of 600 mg q12h intravenously guttae commenced on the 13th day as an alternative to vancomycin(D13-D17). Subsequently, on the 17th day, there was an improvement in hemoglobin level to 78 g/L. However, despite treatment with linezolid, the patient’s fever showed no significant improvement, prompting a switch back to vancomycin at a dosage of 1 g q12h intravenously guttae(D18-D22). On the 21st day, there was a recurrence of gastrointestinal hemorrhage, accompanied by a hemoglobin level of 42 g/L and a PLT count of 224 × 109/L. Gastroscopy revealed the presence of a gastroduodenal ulcer. The patient had no prior history of hemorrhoids, gastrointestinal ulcers, liver cirrhosis, or purpura. Prior to admission, he had not been administered non-steroidal anti-inflammatory drugs (NSAIDs) or steroids. During hospitalization, the only medications given were vancomycin, ambroxol and lidocaine. Additional tests ruled out immunological disorders as the cause of gastrointestinal ulcers, and a positive vancomycin rechallenge test indicated an association between vancomycin and bleeding. After discontinuation of vancomycin, no further bleeding occurred. This case highlights a rare occurrence of vancomycin-induced bleeding without thrombocytopenia, classified as “Certain” according to the World Health Organization-Uppsala Monitoring Centre (WHO-UMC) scale for standardized case causality assessment.ConclusionThis case represents the first documented instance of vancomycin-induced bleeding without thrombocytopenia, as confirmed by a positive rechallenge test. This discovery will aid in the early detection of this rare adverse reaction in future cases.

Myeloid deficiency of heparan sulfate 6-O-endosulfatases impairs bone marrow hematopoiesis

The heparan sulfate (HS) 6-O-endosulfatases or the Sulfs (Sulf1 and Sulf2) are the only known enzymes that can modify HS sulfation status extracellularly and have been shown to regulate diverse biological processes. The role of the Sulfs in bone marrow (BM) hematopoiesis is not known. In this study, we generated a novel mouse line with myeloid-specific deletion of the Sulfs by crossing Sulf1/2 double floxed mice with the LysM-cre line. The LysM-Sulf knockout (KO) male mice exhibited age-dependent expansion of hematopoietic stem cells and the granulocyte-monocyte lineages in the BM, whereas common lymphoid progenitors and B lymphocyte populations were significantly reduced. Although megakaryocytic and erythroid progenitors were not reduced in the BM, the LysM-Sulf KO males suffered age-dependent reduction of red blood cells (RBCs) and platelets in the peripheral blood, suggesting that the production of RBCs and platelets was arrested at later stages. In addition, LysM-Sulf KO males displayed progressive splenomegaly with extramedullary hematopoiesis. Compared to males, LysM-Sulf KO females exhibited a much-reduced phenotype, and ovariectomy had little effect. Mechanistically, reduced TGF-β/Smad2 but enhanced p53/p21 signaling were observed in male but not female LysM-Sulf KO mice. Finally, HS disaccharide analysis via LC-MS/MS revealed increased HS 6-O-sulfation in the BM from both male and female LysM-Sulf KO mice, however, the distribution of 6-O-sulfated motifs were different between the sexes with compensatory increase in Sulf1 expression observed only in LysM-Sulf KO females. In conclusion, our study reveals that myeloid deficiency of the Sulfs leads to multilineage abnormalities in BM hematopoiesis in an age- and sex-dependent manner.

Investigate the association between genetic polymorphisms of ACE and ACE-2 with some biomarkers in Iraqi patients with COVID-19.

BackgroundThe angiotensin-converting enzyme 2 (ACE2) receptor plays a critical role in mediating SARS-CoV-2 infection. Understanding the genetic factors influencing COVID-19 severity is crucial for developing effective treatment strategies. This study aimed to investigate the association between genetic polymorphism of the ACE gene, specifically the insertion/deletion (I/D) polymorphism in the promoter region, and clinical parameters in COVID-19 patients. MethodsA case-control study was conducted with 225 participants from an Iraqi population. Blood samples were collected for hematological analysis and ACE genotyping. The association between ACE genotypes (DD, ID, II), demographic factors, and clinical parameters, including D-dimer, ferritin, lactate dehydrogenase (LDH), C-reactive protein (CRP), white blood cell (WBC) count, lymphocyte count, packed cell volume (PCV), red blood cell (RBC) count, hemoglobin (HB), and platelet count, was assessed. ResultsCOVID-19 patients exhibited significant differences compared to controls regarding D-dimer, ferritin, LDH, CRP, WBC, lymphocytes, PCV, and RBC (P < 0.05), while no significant differences were found for HB and platelet counts. The DD genotype was predominant (43.11 %), followed by ID (39.56 %) and II (17.33 %). Notably, a significant association was observed between D-dimer levels and ACE genotype (P < 0.0001), with higher levels observed in individuals with the DD genotype. Additionally, a significant correlation was found between ACE genotype and sex (P = 0.0163). No significant association was observed between genotype and ICU admission or lung CT severity. ConclusionOur findings suggest a potential link between the ACE D/D genotype and elevated D-dimer levels in COVID-19 patients, indicating a potential role for ACE gene polymorphism in influencing disease severity. Further research is warranted to elucidate the underlying mechanisms and explore the potential for personalized treatment approaches based on ACE genotype.

NBEAL2 gene mutations do not always lead to gray platelet syndrome: A case report.

Gray platelet syndrome (GPS) is a rare disease caused by homozygosity and compound heterozygosity for autosomal mutations on the NBEAL2 gene, which is characterized by a deficiency of platelet α-granules, bleeding symptoms. However, in this study, we report 2 NBEAL2 gene mutations in an easy bruising family without gray platelet and bleeding. A 33-year-old female nurse sought admission to our laboratory due to a tendency to bruise easily and unwell in daily life. However, there are no signs of petechiae or excessive bleeding in her daily life. Coagulation tests, routine blood tests and platelet staining of blood smears were all normal. The whole exome sequencing and Sanger sequencing were used to identify the causative variant of the patient. Furthermore, the morphology of platelets was examined using electron microscopy. Whole exome sequencing revealed the presence of 2 mutations in the NBEAL2 gene: p.Thr365fs and p.Ala310Thr. This prompted the consideration of a GPS diagnosis. However, platelet electron microscopy did not identify any abnormalities, leading to the exclusion of GPS. These 2 NBEAL2 gene mutations (p.Thr365fs and p.Ala310Thr mutations) do not affect the degranulation of platelets.

Baseline Platelet Count Predicts Infarct Size and Mortality after Acute Myocardial Infarction.

Platelets greatly contribute to cardiovascular diseases. We sought to explore the association of platelet counts with infarct size and outcome in patients presenting with acute ST-segment elevation MI (STEMI) treated with primary percutaneous coronary intervention (PPCI). In this retrospective study, we grouped 1,198 STEMI patients into tertiles (T) based on platelet count on admission: T1 = 102-206 [109 platelets/L] (n = 402), T2 = 207-259 [109 platelets/L] (n = 396), and T3 = 260-921 [109 platelets/L] (n = 400). Primary endpoint was 1-year all-cause mortality. Patients with highest platelet counts on admission showed the greatest area at risk and infarct size: area at risk (median) was 22.0% (interquartile range [IQR]: 12.0-39.8%) in T1, 21.0% (IQR: 11.0-37.1%) in T2, and 26.0% (IQR: 14.9-45.0%) of the left ventricle in T3 (p = 0.003); final infarct sizes after 7 to 14 days were as follows: 10.0% (IQR: 2.0-21.0%) in T1, 9.0% (IQR: 2.0-20.7%) in T2, and 12.0% (IQR: 3.0-27.3%) of the left ventricle in T3 (p = 0.015) as serial imaging revealed. At 1 year, 16 all-cause deaths occurred in T1, 5 in T2, and 22 in T3 (log-rank test, p = 0.006). After adjustment, T1 and T3 were associated with all-cause 1-year mortality (T1: hazard ratio [HR] = 3.40, 95% confidence interval [CI] = 1.23-9.54, p = 0.02; T3: HR = 3.55, 95% CI = 1.23-9.78, p = 0.01) compared with T2. At 5 years, all-cause mortality remained numerically higher in the T1 and T3. In patients with STEMI undergoing PPCI, low and high blood platelet levels on admission were associated with increased long-term mortality (Fig. 1).

Immune or inherited thrombocytopenia? A population-based cohort study on children and adolescents presenting with a low platelet count.

Thrombocytopenia is a common hematologic finding in children and adolescents. Immune thrombocytopenia (ITP) is the most common cause of this finding, but the differential diagnosis includes a growing list of genetic disorders. We aimed to report differences in phenotypes of patients with ITP, inherited platelet disorder (IPD)/primary immunodeficiency disorder (PID), and other causes, with a focus on differentiating ITP from inherited thrombocytopenia. This retrospective, population-based observational cohort from 2006 to 2020 involved 506 Finnish children under 16years of age presenting with isolated thrombocytopenia. Of the 506 participants, 79.7% had ITP, 6.7% had IPD/PID, and 13.6% had other causes of thrombocytopenia. A platelet count of ≤12 × 109/L best distinguished between ITP and other reasons with a sensitivity of 60% and a specificity of 80%. Among patients with the lowest platelet count of less than 10 × 109/L, 95.9% had ITP, 3.3% had IPD/PID, and 0.8% had other causes. Severe bleeding events were reported in 20 patients (4.0%), but there were no cases of intracranial or fatal bleeding due to thrombocytopenia. Up to 50% of patients with a high suspicion of inherited thrombocytopenia remained without a specific diagnosis despite genetic testing. ITP remains the most common cause of thrombocytopenia. A platelet count of ≤12 × 109/L often leads to an ITP diagnosis. Genetic disorders are rare but should be suspected in patients with persisting thrombocytopenia, especially with platelet counts constantly above 12 × 109/L, a positive family history, or atypical clinical features.

A-073 Is there a relationship of FIB 4 with weight and insulin resistance in subjects without type 2 diabetes and with normal transaminases?

Abstract Background Metabolic dysfunction-associated steatotic liver disease (MASLD) is a common hepatic disease, affecting 20-30% of the world's population, and is more frequent in subjects with cardiometabolic diseases. The Fibrosis-4 Index (FIB-4) is a non-invasive scoring system that helps estimate the risk of hepatic fibrosis in MASLD. It is based on age, aspartate aminotransferase (AST), alanine aminotransferase (ALT) and platelets (PLT). MASLD is associated with high risk of insulin resistance (IR) and type 2 diabetes (DM2), and subjects with overweight or obesity are more likely to have both. In such cases the association of FIB-4 with IR and weight are evident. It is questioned whether, even in subjects without DM2 and with normal transaminases, FIB-4 could be associated with the body mass index (BMI) and the homeostasis model assessment of insulin resistance index HOMA-IR, which is the index usually used to assess IR. The objective of this study was to evaluate how FIB-4 would behave in subjects without DM2 and with normal transaminases and whether there was a correlation between FIB-4 with HOMA-IR and BMI in these subjects. Methods Authors studied the distribution of FIB-4 results and correlation of HOMA-IR and BMI with FIB-4 using the database of a clinical laboratory. We included patients aged 20-65 years, both gender, who measured fasting glucose and insulin, in the same visit, and transaminases and platelets within up to one year before or later. Were excluded hospitalized subjects, those using medications to treat DM2, dyslipidemia, subjects with blood glucose over 200 mg/dL, and AST and/or ALT above the reference range for gender and age. The methods and reference intervals were: transaminases: UVKinetic, AST &amp;lt; 40 U/L for men and &amp;lt; 32 U/L for women, ALT &amp;lt; 41 for men and &amp;lt; 33 for women; PLT: automated method, 150000-450000/µL. Glucose: hexokinase, 70 - 99 mg/dL; Insulin: electrochemiluminescence, 2-12 mcU/mL. HOMA-IR index was calculated using the equation: glucose (mg/dL) x insulin (mcIU/mL) / 405 and FIB-4 using the equation: age (years) × TGO (U/L) / [(platelets (µL) ÷1000) x √TGP (U/L))]. Shapiro Wilk test was performed to assess whether the analytes had normal distribution. To evaluate the correlation with HOMA-IR, Spearman's non-parametric correlation test was used. Results After applying the exclusion criteria and subtracting outliers 270,135 records were selected, 75% female, with 39,9 ± 10.1 years. As the parameters did not present Gaussian distribution, the results are shown in median and 25th and 75th percentiles: AST=18.9 (16-22); ALT=17.2 (13-23.3); PLT=266000 (229000-308000); glucose = 89.3 (84.2-95); insulin = 8.17 (5.52-12.8); HOMA-IR=1.81 (1.19-2.75); IMC=26.53 (23.80-29.91); FIB-4 = 0.65 (0.5 - 0.86). A weak and surprisingly negative correlation between HOMA-IR and BMI with FIB-4 was found. (HOMA-IR: r=- 0.2, p&amp;lt;0.001; BMI: r=- 0.1, p&amp;lt;0.01). Conclusions Our study showed that in a population with normal transaminases, FIB-4 had results compatible with low risk of fibrosis in subjects without DM2 even in cases with high HOMA-IR or BMI. The negative correlation was surprising and perhaps indicates that FIB-4 may not be a good predictor of liver fibrosis for this population profile.

Key biomarkers in type 2 diabetes patients: A systematic review.

Type 2 diabetes mellitus (T2DM) is not just a local health issue but a significant global health burden, affecting patient outcomes and clinical management worldwide. Despite the wealth of studies reporting T2DM biomarkers, there is an urgent need for a comparative review. This review aims to provide a comprehensive analysis based on the reported T2DM biomarkers and how these are linked with other conditions, such as inflammation and wound healing. A comparative review was conducted on 24 001 study participants, including 10 024 T2DM patients and 13 977 controls (CTL; age 30-90 years). Four main profiles were extracted and analysed from the clinical reports over the past 11 years: haematological (1084 cases vs. 1458 CTL), protein (6753 cases vs. 9613 CTL), cytokine (975 cases vs. 1350 CTL) and lipid (1212 cases vs. 1556 CTL). This review provides a detailed analysis of the haematological profile in T2DM patients, highlighting fundamental changes such as increased white blood cells and platelet counts, accompanied by decreases in red blood cell counts and iron absorption. In the serum protein profile, a reduction in albumin and anti-inflammatory cytokines was noted along with an increase in globulin levels and pro-inflammatory cytokines. Furthermore, changes in lipid profiles were discussed, specifically the decreases in high-density lipoprotein (HDL) and the increases in low-density lipoprotein (LDL) and triglycerides. Understanding the changes in these four biomarker profiles is essential for developing innovative strategies to create diagnostic and prognostic tools for diabetes management.

A-145 Are EDTA and Citrate Collection Devices Interchangeable for Estimating Blood Platelets Values using Sysmex XN™Analyzers ?

Abstract Background In the Sentara Healthcare System patient blood platelets are estimated using Sysmex XN™ analyzers. The blood specimens are collected in devices with EDTA anticoagulant. In the Emergency Departments, upon the first assessment of a patient a number of collection devices, including one with Na citrate anticoagulant, are obtained in case repeated testing is required. To prevent the collection of an additional EDTA device we investigated whether devices with Na citrate anticoagulant could be used interchangeably with EDTA devices for platelets values estimation. Methods Instruments: Sixteen Sysmex XN analyzers (Siemens) located in several institutions. Specimens: Seven-hundred nighty-eight patient blood specimens were obtained with both EDTA and citrate BD™ collection devices. The specimens, platelets numbers between 5000 and 800,000 platelets/mcL, were assayed in parallel within two minutes, after receipt in the laboratory, and then at intervals to reach 20 hours. The data were transferred electronically to Minitab® (version 21, Minitab Inc) statistical software. The paired data were analyzed with orthogonal, ordinary least squares (OLS), weighted polynomial ordinary least squares (WPOLS) and locally weighted scatterplot smoother (lowess) regression models and their graphic representations. The CLIA’s total error criterion (target value ± 25%) was used for evaluating acceptable variability. Results The parallel box plot of the differences between specimens as obtained with EDTA and those as obtained with Na citrate, showed that the distribution of the differences was similar between instruments and within the allowable total error of ± 25%. The orthogonal and the OLS models showed similar regression lines (Orthogonal: Y=3.3+0.94X: OLS: Y=-1+0.93X) and increase in variance for increasing platelets values. Consequently, a WPOLS model was employed for additional analyses. The ANOVA table of the WPOLS showed that there were no probabilistically significant differences between regression lines for either platelet’s values (P = 0.5) or specimen age (P = 0.1). Tridimensional surface plots clearly illustrated that the differences were distributed within -12.5% and the zero lines. The aptness of the WPOLS model was demonstrated by the standardized residual (std. res.) analysis showing quasi-normal distribution, with a few outliers (std. res. &amp;gt; |3|) and two influential observations for (Hi &amp;gt; 0.5) which were for platelets values less than 10,000 platelets/mcL. Conclusions The differences between EDTA and citrated specimens were within zero and -12.5%, this represented half of the CLIA’s criterion. The weighted polynomial regression model was employed due to the heteroscedasticity of the variance, The simultaneous evaluation by platelets values and the age of the specimen did not detect probabilistically significant differences by either platelets values or age of the specimen. This limited study suggested that EDTA and citrate anticoagulated specimens may be used interchangeably for patient care for specimens with platelets values between 10,000 and 800,000/mcL and within twenty hours from collection. However, the limited number of patients precluded to evaluate whether the detected outliers represented group(s) of patients with a particular medical condition and or pharmacotherapeutic treatment. Finally, the electronic collection and transfer of specimens to statistical software, such as Minitab, was crucial for performing this study.

Inherited Predispositions to Myeloid Neoplasms: Pathogenesis and Clinical Implications.

Myeloid neoplasms with and without preexisting platelet disorders frequently develop in association with an underlying germline predisposition. Germline alterations affecting ANKRD26, CEBPA, DDX41, ETV6, and RUNX1 are associated with nonsyndromic predisposition to the development of myeloid neoplasms including acute myeloid leukemia and myelodysplastic syndrome. However, germline predisposition to myeloid neoplasms is also associated with a wide range of other syndromes, including SAMD9/9L associated predisposition, GATA2 deficiency, RASopathies, ribosomopathies, telomere biology disorders, Fanconi anemia, severe congenital neutropenia, Down syndrome, and others. In the fifth edition of the World Health Organization (WHO) series on the classification of tumors of hematopoietic and lymphoid tissues, myeloid neoplasms associated with germline predisposition have been recognized as a separate entity. Here, we review several disorders from this WHO entity as well as other related conditions with an emphasis on the molecular pathogenesis of disease and accompanying somatic alterations. Finally, we provide an overview of establishing the molecular diagnosis of these germline genetic conditions and general recommendations for screening and management of the associated hematologic conditions.

A Predictive Model for Disseminated Intravascular Coagulopathy in Sepsis: An Observational Study.

Sepsis remains a significant global health challenge due to its high morbidity and mortality rates. Disseminated Intravascular Coagulopathy (DIC) represents a critical complication of sepsis, contributing to increased mortality and economic burden. Despite various prognostic scoring systems, there is a lack of a specific model for DIC prediction in sepsis patients. This observational study included 336 sepsis patients. Clinical and laboratory data were collected, and prognoses were defined according to established criteria. We enrolled 336 patients, with 304 in the non-DIC group and 32 in the DIC group. Patients with DIC had notably lower platelet (PLT) and higher levels of prothrombin time (PT), lactate (LAC), and procalcitonin (PCT) compared to those without DIC. Univariate and multivariate analyses identified risk factors associated with the DIC, showing that PLT (OR = 0.985, 95% CI 0.978-0.993, p < 0.001), PT level (OR = 1.140, 95% CI 1.004-1.295, p = 0.044), and LAC (OR = 1.101, 95% CI 0.989-1.226, p = 0.078) were related factors. A risk model was established, and its sensitivity and specificity in predicting DIC among sepsis patients were assessed by comparing it to the SOFA score. The area under the ROC curve for the model was 0.850, while the SOFA score was 0.813. With a model score >-2.12, the sensitivity for predicting DIC was 84.4%, and the specificity was 75.0%. Our study introduces a predictive model for DIC detection in sepsis patients, emphasizing the need for clinicians to focus on patients with high model scores for timely intervention.