Medical Laboratory Journal

Chromatographic insights into hemoglobinopathies: Spectrum analysis by high-performance liquid chromatography in a western Indian tertiary care hospital

Preeti Doshi

Background: Hemoglobinopathies are a group of inherited disorders characterized by abnormal hemoglobin structure or synthesis, primarily classified into thalassemia syndromes and structural hemoglobin variants. Recognized as a global health priority, these disorders necessitate accurate diagnostic approaches. High-performance liquid chromatography (HPLC) has emerged as a reliable method for their detection. This study aimed to analyze the spectrum of hemoglobinopathies using HPLC in a population from Western Maharashtra, India.
Methods: A cross-sectional study was conducted at Bharati Vidyapeeth (DTU) Medical College, Pune, over three years. A total of 1,455 specimens from individuals of both genders were analyzed using the VARIANT™ II β-Thalassemia Short Program based on ion-exchange chromatography. Hematological parameters were assessed for all subjects, and the prevalence of hemoglobinopathies, along with their subtypes, was documented.
Results: The mean age of the study population was 26.91 ± 7.06 years, with a female predominance. The overall incidence of hemoglobinopathies was 8.78%, with β-thalassemia minor being the most prevalent, followed by β-thalassemia major, Hb S trait, and Hb S disease. Rare variants included two cases of Hb D Punjab heterozygosity and one case of Hb E heterozygosity. The Mentzer index exhibited a positive correlation with Hb F and mean corpuscular hemoglobin (MCH) and a negative correlation with hemoglobin levels, Hb A, and red cell distribution width (RDW-C).
Conclusion: The study revealed an 8.78% prevalence of hemoglobinopathies in the region, with β-thalassemia trait (Heterozygous) being the most common. Notably, co-inheritance of Hb E with β-thalassemia exacerbated anemia severity. These findings underscore the importance of HPLC-based screening for early diagnosis and management of hemoglobinopathies in high-risk populations.

A morphological study with clinical, pathological, and immunohistochemical characterization of non-Hodgkin lymphoma

Mohit Kumar

Background: Non-Hodgkin lymphoma (NHL) represents a heterogeneous group of lymphoproliferative malignancies with unique presentation and treatment response. This study was intended to assess the histomorphology of non-Hodgkin lymphoma subtypes and correlate with clinicopathological and immunohistochemical findings.
Methods: This retrospective study was conducted on all specimens diagnosed as NHL by histopathological analysis, with immunohistochemical correlation in the pathology department, for two years. Clinical details such as age, gender, site of the lesion, and Nodal / extranodal presentation were recorded. Histopathological analysis was performed, and Immunohistochemical (IHC) reports were obtained. Further histopathological findings were correlated with IHC results. Statistical analysis was done based on the frequency distribution.
Results: This study includes 48 cases. Most cases are 61 to 70 years old, with a male predominance (56.25 %). The most common clinical diagnosis was lymphoma (79.17 %). Most of the lesions were of lymph nodal origin (60.42 %). The cervical group (35.42 %) is the most common lymph node affected, followed by the axillary nodes. The most common extranodal sites are the retroperitoneum and stomach. On histopathological evaluation, the most common diagnosis was NHL (68.75 %) without further subtyping. Among cases where subtyping was done, the most common lesion were follicular lymphoma and diffuse large B-cell lymphoma (DLBCL). On IHC evaluation, B cell neoplasms (85.42%) were common compared to T cell neoplasms (14.58 %). The most common subtype was DLBCL (52.08 %), followed by follicular lymphoma (16.67 %).
Conclusion: Our study found diffuse large B cell lymphoma (DLBCL) as the most common type of Non-Hodgkin’s Lymphoma. Cervical lymph nodes were found to be the most common site of involvement. But the involvement of rare sites like the testis and palate was also found. Hence the probability of NHL in these rare sites should always be considered.

SARS-CoV-2 IgG antibody response to a viral vector vaccine among health care workers in a tertiary care hospital

Kavitha Paul Konikkara

Background: The surge of COVID-19 has weighed heavily on health-care systems and hospitals. Health-care workers are at high risk of exposure both in the community and workplace when providing care to patients. Quantitative assays to detect antibodies against SARS COV-2 help to determine individual antibody titer and in longitudinal monitoring of antibody response. The present study was undertaken as there is not much data available regarding the presence of SARS COV-2 antibodies among health care workers in Kerala. The objective was to estimate the IgG antibody response and to find out the associated factors in vaccinated health-care workers.
Methods: A cross-sectional study was conducted among 187 vaccinated health- care workers. The blood sample was collected from health care workers and quantitative determination of humoral antibodies (Including IgG) was done by Chemiluminescence immunoassay. The antibody response was correlated with various factors. Analysis was done using the software IBM SPSS Version 25.
Results: The seroconversion rate following the Covishield vaccine was found to be 99.47% with mean, median, and standard deviation of antibody titer being 1291.0, 311.40 and 1792.500, respectively. Statistical significance in antibody titer was observed among different categories of health care workers and in those with previous COVID-19 infection. Higher titer was noted within 3 months of vaccination and thereafter waning of titer was seen which was also statistically significant.
Conclusion: No correlation was observed with age, gender, co-morbidities, area of work and Body Mass Index (BMI). Vaccination plays a major role in containing the infection. The immune response among vaccinated health-care workers was satisfactory.

Incidence of phototherapy-induced hypocalcemia in neonates with hyperbilirubinemia

Poonam Wade

Background: Hyperbilirubinemia is the most common clinical abnormality in newborns during the first week of life. Phototherapy is the standard treatment but may cause complications, including hypocalcemia, a lesser-known yet significant adverse effect. This study investigated the impact of phototherapy on serum calcium levels in neonates with hyperbilirubinemia.
Methods: This observational study was conducted in the NICU of BYL and T.N. Medical College, Mumbai, over 12 months after ethical approval. We included 100 neonates (> 34 weeks gestational age) with hyperbilirubinemia receiving phototherapy. Serum calcium levels were measured before and after phototherapy, and neonates were monitored for clinical hypocalcemia. Data were recorded in a pre-formatted proforma and analyzed using SPSS version 27 (p <0.05 was considered statistically significant).
Results: The mean age of neonates was 3±1.2 days, the mean gestational age was 37±1.2 weeks, and the mean jaundice duration was 2.8±1.10 days. Median serum calcium levels decreased significantly from 8.6 mg/dL (8.49-8.61) pre-phototherapy to 7.35 mg/dL (7.33-7.50) post-phototherapy (p <0.001). Hypocalcemia (serum calcium ≤7 mg/dL) occurred in 16% of neonates, with 60% exhibiting mild symptoms like jitteriness or irritability.
Conclusion: Phototherapy significantly reduces serum calcium levels in jaundiced neonates, with a notable incidence of hypocalcemia. Clinicians should monitor calcium levels and watch for symptoms during phototherapy to prevent complications. Further studies are needed to establish preventive strategies.

Bronchopulmonary lophomoniasis in a prisoner: A case report

Mohammad Hadi Tajik Jalayeri

Background: Bronchopulmonary lophomoniasis, an emerging yet frequently overlooked respiratory infection, is caused by the flagellated protozoan Lophomonas blattarum (L. blattarum). Although predominantly observed in immunocompromised individuals, its occurrence in immunocompetent hosts, particularly within high-risk settings like correctional facilities, is uncommon.
Case Presentation: This report details the case of a 47-year-old male prisoner from Golestan Province, Iran, who presented with a four-month history of chronic cough, dyspnea, and purulent sputum. The initial diagnostic workup, including blood and sputum cultures, yielded no evidence of bacterial or fungal infection. However, microscopic examination of bronchoalveolar lavage (BAL) fluid revealed L. blattarum trophozoites, leading to a definitive diagnosis of lophomoniasis. Of particular interest was the elevated serum immunoglobulin E (IgE) level (387 kU/L; normal range <160 kU/L), which may indicate an underlying allergic predisposition or a concurrent parasitic infection. The patient's symptoms resolved completely after a four-week regimen of metronidazole.
Conclusion: This case highlights the importance of considering lophomoniasis in the differential diagnosis of chronic respiratory symptoms, especially in settings with poor hygiene. For accurate diagnosis in similar patient populations, a heightened clinical suspicion combined with BAL microscopy are essential.

The effect of cigarette smoking on serum homocysteine, folic acid, and vitamin B12 concentrations in patients with cardiovascular diseases

Mohsen Tatar

Background: Smoking is recognized as a significant risk factor for cardiovascular diseases (CVD), yet its influence on blood levels of homocysteine, folic acid, and vitamin B12 is not well understood. This study aimed to explore how smoking affects these biochemical markers in patients with CVD.
Methods: The study included 88 participants diagnosed with CVD, who were categorized into smokers (n=44) and non-smokers (n=44). Serum concentrations of homocysteine, folic acid, and vitamin B12 were assessed using ELISA. Additionally, blood pressure (both systolic and diastolic) and body mass index (BMI) were recorded.
Results: Smokers showed significantly lower levels of folic acid (22.41 ± 5.95 ng/mL) compared to non-smokers (28.05 ± 4.13 ng/mL, p = 0.000). No significant differences were observed in homocysteine (p = 0.958) or vitamin B12 (p = 0.578) levels between the two groups. A negative correlation was found between folic acid and systolic blood pressure in smokers, while no significant associations were noted among folic acid, vitamin B12, and homocysteine.
Conclusion: In patients with CVD, smoking is linked to significantly lower folic acid levels, which may lead to increased systolic blood pressure. These results underscore the need to monitor folic acid levels in smokers who are at risk for cardiovascular issues.

The effect of an interval training course on nitric oxide levels, insulin resistance, and some blood lipid factors in type 2 diabetic male rats

Roohollah Mohammadi Mirzaei

Background: Diabetes is a metabolic and vascular disorder characterized by endothelial dysfunction. Physical activity, particularly intermittent exercise, may offer therapeutic benefits. This study examined the effects of six weeks of intermittent exercise on serum nitric oxide (NO), insulin resistance, and lipid profiles in male diabetic rats.
Methods: Sixty Wistar rats (8 weeks old, 200 ± 20 g) were divided into six groups (n=10): healthy control, sham, interval training, interval training + saline, diabetic control, and diabetic + interval training. Diabetes was induced via nicotinamide-streptozotocin injection. The exercise groups underwent treadmill training (5 sessions/week for 6 weeks). Post-intervention, glucose, insulin, NO, LDL, HDL, triglycerides, and cholesterol were measured.
Results: Diabetes induction significantly increased glucose and insulin resistance while reducing insulin and NO levels compared to controls (P < 0.001). After six weeks, the diabetic + exercise group showed significant reductions in glucose and insulin resistance (P < 0.001) and increased NO levels (P < 0.001) versus the diabetic group. Insulin levels did not differ significantly among the groups (P = 0.11). Lipid profiles (LDL, HDL, triglycerides, cholesterol) remained unchanged (P > 0.05).
Conclusion: Six weeks of intermittent exercise improved glucose metabolism and vascular function in the diabetic rats by reducing insulin resistance and increasing NO levels, suggesting its potential as a non-pharmacological therapy for diabetes-related endothelial dysfunction.

JAK2 mutation, a potential cause of increasing Hemoglobin levels in symptomatic aged women

Mohammad Taher Hojjati

Background: Polycythemia Vera (PV) is a myeloproliferative neoplasm (MPN) characterized by the overactivity of erythroid progenitors, leading to excessive red blood cell (RBC) production. More than 90% of PV cases harbor a Janus kinase 2 (JAK2) gene mutation. This study aimed to assess the prevalence of JAK2 mutations in individuals with elevated hemoglobin (Hb) levels referred to the laboratory by physicians.
Methods: In this descriptive cross-sectional study, genomic DNA from 72 patients was analyzed for JAK2 mutations using a TaqMan-specific probe.
Results: Of the 72 patients, 24 (33.3%) were women and 48 (66.6%) were men. JAK2 mutations were detected in 33 cases (45.5%), while 39 (54.2%) were negative. Notably, 15 of 24 female patients (62.5%) tested positive for the JAK2 mutation, compared to 18 of 48 male patients (37.5%).
Conclusion: Our findings suggest that screening for JAK2 mutations is particularly important in women with above-normal Hb levels.

Attenuation of glucose mediated DNA glycation by Tamarix aphylla extract

Parisa Hasanein

Background: DNA glycation, a process where Glc non-enzymatically binds to DNA, is implicated in various detrimental effects, including strand breaks, mutations, and altered gene expression. This damage is considered a significant contributor to the pathogenesis of diabetes mellitus and its associated complications. Consequently, there has been increasing interest in identifying antiglycation agents as a strategy for preventing and mitigating these complications. Prior research has indicated that extracts from Tamarix aphylla (T. aphylla) leaves possess antidiabetic properties. Therefore, this study aimed to investigate, for the first time, the impact of T. aphylla extract on Glc-mediated DNA glycation.
Methods: DNA samples were incubated with Glc over a four-week period. Subsequently, the modulatory effects of T. aphylla on Glc-induced DNA structural alterations were investigated employing a range of analytical techniques. These methodologies encompassed ultraviolet-visible (UV-Vis) spectroscopy, fluorescence spectroscopy, circular dichroism (CD) spectroscopy, and agarose gel electrophoresis.
Results: The results obtained from UV–Vis and fluorescence spectroscopy demonstrated that T. aphylla extract led to a reduction in the formation of DNA-advanced glycation end products (AGEs). Furthermore, CD spectroscopy and agarose gel electrophoresis analyses indicated that the structural alterations of glycated DNA were diminished in the presence of T. aphylla extract.
Conclusion: Based on the evidence presented, T. aphylla demonstrates protective properties against DNA glycation. Consequently, pending further rigorous investigation, it may represent a potentially valuable therapeutic agent for mitigating the detrimental consequences of glycation, particularly in environments characterized by elevated Glc concentrations and hyperglycemic states.

Chronic basophilic leukemia; A rare myeloproliferative disorder and the role of flow cytometry for its diagnosis

Esmaeil Shahabi Satlsar

Chronic basophilic leukemia (CBL) is a rare disorder and according to the published data, few cases have been reported as primary CBL. Morphologic findings in CBL mostly mimic chronic eosinophilic leukemia and basophils have unusual and dysplastic morphology. A 47-year-old patient was referred to the hospital with gastrointestinal symptoms and bloating. Complete blood count (CBC) showed leukocytosis and marked eosinophilia. After evaluating the peripheral blood smear (PBS), abnormal leukocytes with hypersegmented nuclei along with an increase in eosinophils were observed. For further investigation, the patient was referred to the hematology clinic and underwent bone marrow aspiration and biopsy. After the morphological examination, flow cytometry was performed on the aspiration sample to accurately diagnose the disease. Flow cytometric findings were in favor of CBL. Since morphological findings are unreliable for basophil detection and definite diagnosis, flow cytometry is a reliable method for the precise detection of basophils, especially in CBL.