Clinico-Hematological Profile and Management of Children With Non-Transfusion Dependent Thalassemia (NTDT) at a Pediatric Center in Northern India

Objective: To study the clinico-hematological profile, complications, and management of children with non-transfusion dependent thalassemia (NTDT) in northern India. Method: We retrieved and analyzed the data of 69 children with NTDT diagnosed between January, 2006 to December, 2018, aged under 18 years from our unit’s records. Result: The participants mean (SD) age was 4.4 (3.1) years, and they presented with anemia (29%), jaundice (13%), hemolytic facies (13%), splenomegaly (87%), thromboembolism (2.9%) and pathological short stature (28.5%). The most common cause of NTDT was ?-thalassemia (45%), followed by either compound-heterozygous or homozygous for E?-thalassemia mutation. The most frequent single genotype observed was compound heterozygous for IVS1-5 (G>C) and codon 26 (G>A). The mean (SD) follow-up duration was 3.5 (2.4) years. On follow-up, 27 children (%) remained transfusion free, and 30 (%) needed occasional transfusions. 63% of patients initially presenting with pathological short stature showed improvement in growth. Amongst children older than 10 years (n=20), subclinical hypothyroidism was detected in 6 children and impaired glucose tolerance test in 1 child. Conclusion: Eß-thalassemia was the commonest cause of NTDT in this population.

Abnormal semen parameters among males in infertile couples: a cross sectional study from a tertiary care centre

Background: In India, the prevalence of primary infertility ranges from 3.9% to 16.8%. Male factor contributes 40-50% of this. Male factor infertility is indicated by decreased sperm concentration, reduced motility, vitality or abnormal sperm morphology. Semen analysis is the single most important investigation to detect male factor infertility. The aim of this study was to analyse the prevalence of abnormal semen parameters among males in infertile couples and their association with contributing factors.Methods: This cross-sectional hospital-based study was carried out in the Department of Pathology at Lady Hardinge Medical College and Smt. Sucheta Kriplani Hospital. A total of 400 cases were analyzed during a period of 6 months. Detailed history of the couple was taken. Semen analysis was done using automated semen analyzer (SQA-vision) after 3 days of abstinence according to the WHO 5th edition 2010 guidelines. The results were analysed using excel sheet and SPSS software.Results: In the present study, 122 cases (30.5%) out of 400 cases had abnormal semen parameters. Most common abnormality detected was asthenozoospermia (14.3%) followed by oligozoospermia (13.8%), azoospermia (10.5%) and teratozoospermia (10.5%). There was significant association of alcohol intake, obesity and trauma with abnormal semen parameters.Conclusions: Asthenozoospermia was the most common abnormality noted in this study. Lifestyle modifications along with timely medical attention in male partners of infertile couples can improve the semen quality.

Clinico-hematological study of abnormalities of platelet count in children with iron deficiency anemia

Background: Iron deficiency anemia is a major cause of morbidity in developing countries like India. The aim of the study was to assess abnormalities of platelet count in iron deficiency anemia and to relate the severity of thrombocytosis with severity of anemia and its association with erythropoietin (EPO) level.Methods: A prospective observational study comprising of 200 children below 18 years confirmed to have IDA. Erythropoietin (EPO) level was done in patients who had thrombocytosis. Degree of thrombocytosis was correlated with EPO and also with ferritin, haematological indices like hemoglobin and MCV (mean corpuscular volume) and blood counts were followed up while on iron therapy for one month.Results: Thrombocytosis was noted in 24.5%. In 75.5% thrombocytosis was mild. Platelet had negative correlation with Hb (hemoglobin). EPO was elevated in 67.35% of thrombocytosis. EPO showed negative correlation with Hb and Ferritin and positive correlation with platelet however, these were non-significant. All patients were treated with standard preparation of ferrous fumarate (33mg elemental iron every 5 ml) in a dose of 3mg/kg/day of elemental iron along with appropriate dietary advice.' On one month follow up 92% of the study population showed normalization of platelet count.Conclusions: Nearly One-fourth of children had thrombocytosis. Platelet count was inversely related to Hb and ferritin level. EPO was increased in two-third cases of thrombocytosis and showed positive correlation with platelet count. As authors excluded patients with severe IDA requiring blood transfusion, authors did not get any thrombocytopenia in present study.

Efficacy of Ketamine and Fentanyl Vs. Intrathecal Bupivacaine Post-operatively in Patients Undergoing Abdominal Hysterectomy: A Prospective Comparative Study

Background: Although disturbing emergence reactions are associated with its use, ketamine has several clinically useful properties, including analgesia and less cardiorespiratory depressant effects than other anaesthetic agents, in fact it causes some stimulation of the cardiovascular system. Methods: Two group were included in this study i.e. Group A & Group B. Both group had 18 cases. This study conducted in Department of Anaesthesiology & Critical Care, Mahatma Gandhi Medical College & Hospital. Results: Demographic variables were compared between both groups A and B. Group A is A significant faster onset of sensory block was found in ketamine group in comparison to fentanyl group. (p-value<0.001). While the time to achieve the highest level of sensory block was found to be almost similar in both the groups. Conclusion: Addition of ketamine or fentanyl to spinal bupivacaine were equally effective in pain control after abdominal hysterectomy.

Hereditary Elliptocytosis: An Incidental Finding in a 6-year-old Child.

A case of hereditary elliptocytosis (HE) in a 6-year-old child diagnosed as an incidental finding has been presented.

Preliminary Study of Hyphomycetes of Central India.

During the preliminary investigation of hyphomycetes from central India eight species of freshwater and freshwater-borne Hyphomycetes encountered in foam samples collected from the different streams and rivers. These fungi namely viz., Wiesneriomyces laurinus Kirk, Flabellospora multiradiata Nawawi, Isthmotricladia gombakiensis Nawawi, Triscelophorus acuminatus Nawawi, Flabellospora acuminata Descals and Webster, Sporidesmium leonense M.B. Ellis, Acrodictys globulosa Toth M.B. Ellis, and Acladium phoenicis Ramachar are being reported for the first time from central India. Brief notes and illustrations are also provided along with geographical distributions of these fungi in India.

Hemoglobin Agenogi ‑ A rare abnormal beta globin chain variant.

Haemoglobin (Hb) Agenogi is clinically asymptomatic, rare β‑globin chain variant characterized by a substitution of glutamic acid by lysine at position 90 of β‑chain. It elutes in the C‑window on high‑performance liquid chromatography (HPLC). We report a 10‑year‑old male with easy fatigability, lethargy, pallor, and mild splenomegaly. Hematological parameters revealed microcytic hypochromic anemia and mildly raised red blood cells count, suggestive of thalassemia trait. On HPLC, a predominant peak was observed in the C‑window (82.6%) along with raised HbA2 level (9.3%). Based on these findings, a possibility of HbC disease/β‑thalassemia trait doubly heterozygous was considered. Family studies were advised. HPLC findings in father were suggestive of β‑thalassemia trait, while both his mother and brother had an abnormal peak in the C‑window of 42.7% and 40.8%, respectively, with elevated HbA2 values of 5% and 4.9%, respectively. Direct DNA sequencing revealed intervening sequences 1–5 (G → C) in father, confirming β‑thalassemia trait. His mother and brother had heterozygous gene mutation at codon 90 of β‑globin chain (G → A) suggestive of Hb Agenogi. The child carried mutations for both β‑thalassemia trait as well as Hb Agenogi.

Determinants of Nutritional Anemia in Adolescents.

Objectives: To associate the severity of nutritional anaemia with serum levels of ferritin, vitamin B12 and folate; and to determine demographic, socio-economic and nutritional correlates for nutritional anemia in adolescents. Methods: Cross-sectional hospital-based study among 200 adolescents (10-18 y) with anemia. Dietary intake (24-h recall), and serum levels of folate, vitamin B12 and ferritin were estimated. Results: Iron, folate and vitamin B12 deficiency was present in 30.5% 79.5% and 50% of adolescents, respectively. Statistically significant association was observed between severity of anemia and serum vitamin B12 levels, iron intake, folate intake, Vitamin B12 intake, vegetarian diet, attainment of menarche and history of worm infestation. Conclusions: Folate and vitamin B12 deficiencies are more common than iron deficiency in anemic adolescents. Low dietary intake of these nutrients seems to be a significant determinant of their deficiencies.

What regulates hepcidin in poly-transfused β-Thalassemia Major: Erythroid drive or store drive.

Background: Hepcidin, a key regulator of iron homeostasis, is increased by iron overload and infl ammation while suppressed by hypoxia. In spite of iron overload in β-Thalassemia Major (β-TM), a paradoxical decrease in hepcidin is observed. Aim: To assess the opposing effects of enhanced erythropoiesis due to anemia and iron overloading on hepcidin in β-TM patients. Setting and Design: This prospective observational study was done at our tertiary care hospital. Materials and Methods: Eighty-three pediatric polytransfused (> 20 transfusions) patients of β-TM were compared with 70 children who served as controls. Serum assays for ferritin, transferrin receptors (sTfR) and hepcidin were performed. Statistical analysis: Independent Student t test was used to compare variables between both the groups. A Pearson correlation coeffi cient was used to fi nd any correlation between ferritin, sTfR and hepcidin. Results: The mean value of hepcidin in β-TM children was 13.88±10.68 ng/ml (range, 0.9-60 ng/ml) and showed signifi cant negative correlation with sTfR (r = –0.296, P < 0.0066). However, there was no correlation of hepcidin with ferritin. Ferritin and sTfR were signifi cantly elevated in β-TM children compared to controls (P < 0.001). The mean serum hepcidin/ferritin index in the study group (0.00552) was signifi cantly lower (P value < 0.001) than the controls (0.378) thus indicating inappropriate levels of hepcidin to iron overload. Conclusion: In polytransfused β-TM children increased iron demand dominates over iron overload in regulating hepcidin. In spite of excessive iron load, the inappropriate hepcidin levels may further contribute to iron overload enhancing iron toxicity.

Coagulation inhibitors and activated protein C resistance in recurrent pregnancy losses in Indian women.

Background: Thrombophilias, both acquired and inherited, have been investigated in the etiopathogenesis of unexplained recurrent pregnancy loss. Aim: To study coagulation inhibitors and activated protein C resistance (APCR) in recurrent pregnancy losses (RPL) occurring in second and third trimesters. Materials and Methods: A total of 30 pregnant women (group A) with two or more recurrent unexplained fetal loses were evaluated for APCR, protein C deficiency, protein S deficiency, antithrombin deficiency, and antiphospholipid antibodies (APLA). Thirty age-matched controls were taken (group B) comprising of pregnant women with at least one live issue. Statistical Analysis: Comparisons between two group frequencies and group means were made using Chi square test and Student's t test, respectively. Results: Protein C and protein S levels were reduced in group A compared with group B and the difference was statistically significant (P=0.005 and P=0.032, respectively). The mean value of antithrombin was slightly reduced in group A compared with group B. APCR was observed in 16.6% cases and 3.3% controls. However, the difference was not statistically significant. APLA was observed in 20% cases and none of the controls. Of these, lupus anticoagulant was positive in 16.6% cases and anticardiolipin antibodies in 10% cases. Combined defects were seen in seven patients. Conclusion: There is a significant risk of RPL in pregnant women with thrombophilias. Therefore, screening for thrombophilias may be justified in pregnant women with unexplained recurrent fetal wastage, especially in second and third trimester.

Childhood acute erythtoleukemia diagnosis by flow cytometry.

Acute erythroid leukemia in children is very. Here is a case of erythroleukemia in a child of a age 1.5 years, which was diagnosed on peripheral smear, bone marrow examination, cytochemistry but was confimed on immunophenotyping. CD45 versus side scatter demonstrated blast population (29%) expressing CD45 of variable intensity (dim to negative). The myeloid nature of blast population showed bright expression of CD13, CD33. These blasts also showed bright positivity of CD71 which showed erythroid nature of blasts. Flow cytometry can be comprehensive enough to completely subtype cases of leukemias/myelodysplastic syndromes, polycythemia rubra vera, non-neoplastic conditions like reactive erythroid hyperplasia following immunosuppressive therapy or viral infections or nutritional deficiencies, unlyzed RBCs or thrombocytosis which may mimic acute erythroid leukemia on flow cytometry.

Critical evaluation of peripheral smear in cases of anemia with high mean corpuscular hemoglobin concentration in children: A series of four cases.

Mean corpuscular hemoglobin concentration (MCHC), a parameter that is reported as a part of a standard complete blood count by automated analyzer, is a measure of the concentration of hemoglobin in a given volume of packed red blood cell. Values of MCHC significantly above reference range are not physiologically possible due to limitations on solubility of hemoglobin. The high MCHC can give us a clue to certain type of hemolytic anemia and necessitate critical evaluation of peripheral smear to reach a definitive diagnosis. Here we are presenting a series of four cases with raised MCHC, emphasizing the importance of systematic and meticulous examination of the peripheral smear to render a definitive diagnosis.

Clinicohematological study of thrombocytosis.

Objective. To find out etiology and clinical course of thrombocytosis in Indian pediatric population. Methods. A total of 250 patients having thrombocytosis (defined as platelet count >500 x 109/L) on haematological investigations were studied over one yr period. All patients were evaluated clinically and were subjected to investigations, including complete blood counts (CBC) with peripheral smear examination. To elucidate the possible role of inflammatory cytokines in pathogenesis of RT, levels of Interleukin-6 (IL-6) and C - reactive protein (CRP) were estimated. Results. Infants and young children (<2 yr age) were most common group, contributing 60% of total cases. Out of total 250 cases, only 3 (1.2%) cases were found to have primary thrombocytosis and remaining 98.8% cases were having RT. Among RT patients, infections (alone or in association with iron deficiency anemia) were most common cause, accounting for 65% cases, while iron deficiency anemia (IDA) was second most common cause accounting for 41.3% cases (12.6% IDA alone and 28.7 % in association with infections). Other causes included nutritional dimorphic anemia and patients on treatment for megaloblastic anemia, acute lymphoblastic leukemia (during treatment) and lymphoma. Among various groups of RT, IL-6 and CRP levels were higher in patients with infection with or without IDA than IDA alone. One child with essential thrombocytosis and one child with RT had thrombotic complications. On follow up, platelet counts normalized in most of the patients with treatment of underlying conditions. Conclusions. Results of this study suggest that essential thrombocytosis is extremely rare in children. Infections and IDA (alone or in association with infections) are common causes of RT. IL-6 and CRP levels are increased in patients with RT, to a higher level in patients with infections than in patients with IDA. Most patients with RT have uneventful recovery of platelet counts to normal range with treatment for underlying condition.

Evaluation of serum transferrin receptor and sTfR ferritin indices in diagnosing and differentiating iron deficiency anemia from anemia of chronic disease.

Objective. The present study was conducted to assess the utility of serum transferrin receptor (sTfR) and sTfR ferritin indices to differentiate ACD from IDA and also to diagnose coexisting IDA and ACD. Methods. The study group comprised of 30 IDA patients, 30 cases of ACD and 30 age and sex matched controls. Complete hemogram with peripheral smear examination, markers of ACD, iron profile including serum ferritin and serum transferrin receptor levels were done in all patients and controls. Serum TfR and ferritin indices were calculated. Results. sTfR levels were significantly higher in the IDA group compared to ACD group (p<0.001). ACD group was further subdivided into two groups on the basis of sTfR levels (B1<3 μg/ml and B2 ≥ μg/ml), suggesting coexisting IDA in group B2. sTfR/log ferritin index was > 1.5 in all cases of IDA and ACD with coexisting IDA while all pure ACD cases and control subjects had sTfR/log ferritin index < 1.5. All case in IDA group had log sTfR/serum ferritin index > 2.55 and all patients with ACD with or without associated iron deficiency had log sTfR/serum ferritin ratio < 2.55. Conclusion. The sTfR levels along with the above mentioned indices can be very useful in differentiating pure IDA, ACD and ACD with coexisting iron deficiency, thus providing a noninvasive alternative to bone marrow iron.

Acute erythroid toxicity in visceral leishmaniasis: a rare complication of antimonial therapy.

The spectrum of side-effects of sodium stibogluconate is well described, however, little is known regarding the acute erythroid toxicity caused by this drug. We hereby present a case with this unusual complication of antimonial therapy. A 6-year-old male with leishmaniasis was started on parenteral sodium stibogluconate. During the course of treatment, his hemoglobin (Hb) dropped from 7.2 g/dl to 3.5 g/dl. Bone-marrow aspirate showed karyorrhexis in many erythroid precursors with several Leishmania donovanii bodies. Sodium stibogluconate was stopped and amphotericin-B was started. Four days after the cessation of the antimonials, the patient's Hb improved to 5 gm/dl with a corrected reticulocyte count of 10% indicating bone-marrow erythroid regeneration. The exact mechanism of this acute erythroid toxicity of sodium stibogluconate remains unexplored.

Antithymocyte globulin and cyclosporin in children with acquired aplastic anemia.

OBJECTIVE: To assess the responses to ATG and cyclosporin combination in patients of aplastic anemia. METHODS: Twenty three (17M: 6F) patients of aplastic anemia (11 very severe aplastic anemia (VSAA) and 12 severe aplastic anemia (SAA), were administered antithymocyte globulin and cyclosporin. RESULTS: The median age of patents was 8 years (range 6-12 years). Three patients died within 2 months of therapy. Twenty children (11 SAA and 9 VSAA) were finally analysed. Six months after the start of treatment, 8/20 (40%) patients responded-2 complete (CR) and 6 partial responses (PR). At the end of 1 year; 2 patients maintained CR and seven patients continued PR (overall responders 45%). The response was better in SAA (54.5%) with 2 CR and 4 PR; than in VSAA (33%) with 3 PR . Eleven (55%) children were alive without response. One patient developed AML 13 months later. CONCLUSION: We conclude that antithymocyte globulin and cyclosporin combination is an effective treatment for aplastic anemia patients who are ineligible for bone marrow transplantation.

Developmental and hyperthermia-induced expression of the heat shock proteins HSP60 and HSP70 in tissues of the housefly Musca domestica: an in vitro study

The expression pattern of two major chaperones, the heat shock proteins (HSPs) HSP60 and HSP70 was studied in vitro in tissues of the housefly Musca domestica during larval and adult stages of development to identify their immunological relatives and understand their functional significance in normal cellular activities and during thermal stress. Fluorographs of labeled polypeptides and western blots demonstrated that both HSPs are expressed constitutively and heat-induced in all the larval and adult cell types examined. The pattern of whole tissue immunocytochemical staining using anti-HSP60 and anti-HSP70 antibodies corresponded well with the observations from western blots or fluorographs. In developing oocytes, both constitutive and heat inducible expression of HSP60 were regulated in an oocyte stage-specific manner. In unstressed ovaries the expression of these proteins was less pronounced in early stage oocytes (1st - 8th) than at later stages (9th and onward). The heat shock, however, induced both HSP70 and HSP60 to a significantly high level in early stage oocytes (1st-8th) as compared to their respective controls. Our findings indicate the involvement of the HSP60 and HSP70 proteins in the development, growth and differentiation of both somatic and germ line tissues. Furthermore, the enhanced co-expression of HSP70 and HSP60 upon heat shock in various larval and adult cell types suggests the possible role of HSP60 in thermoprotection.