Does Antioxidant Superoxide Dismutase levels Decline with Advanced primary Knee Osteoarthritis in Synovial Fluid ? A Pilot Study

Introduction: Osteoarthritis (OA) is a progressive, degenera?ve disease that leads to joint pain, tenderness, s?ffness, locking, effusion, reduced mo?on, swelling, crepitus, and disability. The pain in OA is the most significant clinical feature and impacts func?on, mobility, quality of life, and the reason for medical advice. Methods: Fi?y individuals with primary knee OA in the age range of 45–90 years were chosen at random for the research (N=50). The American College of Rheumatology’s diagnos?c criteria were employed to diagnose osteoarthri?s, and a visual analogue scale was u?lized to score the severity of pain. Knee OA was graded using the Kellgren-Lawrence (K-L) radiographic assessment method. The an?oxidant levels of superoxide dismutase in the synovial fluid were measured by using a spectrophotometric assay. Results: Grades 1, 2, 3, and 4 have SOD ac?vity values of 1.43±0.55, 1.44±0.72, 0.92±0.52, and 0.87 ±0.52 U/ml, respec?vely, in synovial fluid. Synovial fluid SOD ac?vity was higher in grades 1 & 2 of KOA as compared to grades 3 &4 and the difference was sta?s?cally significant (p<0.05). Conclusions: There was a link between K-L grade and synovial an?oxidant ac?vity level. In the late stages of knee osteoarthri?s, the an?oxidant enzyme (SOD) ac?vity was reduced.According to the results of this study, regular an?oxidant supplementa?on to early osteoarthri?s pa?ents may delay disease progression by improving the an?oxidant status of the knee, which neutralises free radicals and thus prevents car?lage damage.

Interleukin 6 and disease transformation in chronic myeloid leukemia: A Northeast Indian population study

Objective: Growth factor independence 1 (GFI1), a transcriptional repressor, is required for hematopoietic stem cell maintenance and self-renewal in addition to controlling differentiation and proliferation of myeloid cells. As murine studies have demonstrated that this transcription factor has a notable role in the initiation and progression of acute myeloid leukemia (AML) disease, the aim of the current study was to investigate and review the influence of GFI1 in human AML cells. Methods: GFI1 expression levels were measured by means of real-time polymerase chain reaction in 96 primary AML samples which were then compared to gene expression levels observed in 18 healthy subjects. Moreover, GFI1 expression patterns were analyzed based on specific AML subtypes including acute promyelocytic leukemia (APL). Finally, leukemic cells were stained to measure levels of myeloperoxidase (MPO) activity. Results: This study reports that AML patients have significantly higher GFI1 mRNA levels in comparison to healthy subjects and that, when considering AML subtypes, patients with APL have higher GFI1 expression than non-APL patients. Conclusion: It is also concluded that GFI1 overexpression in patients with high MPO levels, such as those of the APL subtype, is correlated with favorable disease prognosis as supported by other studies which demonstrate that increased peroxide activity and GFI1 are independently correlated with a favorable prognosis

Adenosine Deaminase as a Biomarker for the Diagnosis of Tubercular Pleural Effusion

Background:The aim of our study was to study and compare the levels of Adenosine deaminase, total proteins and glucose levels in tubercular and non tubercular Pleural Effusion patients, and simultaneously to establish the diagnostic utility of Pleural fluid adenosine deaminase activity in tubercular pleural effusion. Study Design: A hospital based cross sectional study was conducted on patients attending the Out Patient Department or admitted in Medicine or Pulmonary Ward of SMIH, Dehradun (UK) for a period of 6 months from Aug2017 to Jan 2018. 56 cases of tubercular pleural effusion above 15years were selected randomly for the study. The control group consists of 54 subjects of non tubercular pleural effusion. Exclusion criteria was age below 14 years & those who were not willing to participate. Inclusion criteria was age above 14 years, Pleural fluid smear positive for AFB or fluid culture positive for mycobacteria tuberculosis, positive sputum culture, positive pleural biopsy. All the cases and controls were subjected to complete history, clinical examination and pleural fluid estimation for ADA, protein and glucose. Material & Method: Both the cases and controls were subjected to complete history taking, clinical examination and pleural fluid estimation for ADA, protein and glucose. The pleural fluid sample was processed on Vitros 250 for glucose & protein and on semi autoanalyzer (Erba chem) for ADA.Results & Conclusion: In the test group the Mean ± SD of ADA activity was 103.29±61.26 IU/l whereas in control group it was25.0±18.2 IU/l. Thus the activity of ADA in the test group was significantly higher than the control group (p<0.0001). In the test group, the Mean ± SD of glucose level was 65.80±29.27 mg/dl and in the control group was 123.1±79.9 mg/dl. Thus the level of glucose in the control group was higher than the test group (p<0.0001). Similarly the mean values of protein were found to be much higher in test group (4.69±0.97 gm/dl) as compared to the control group (3.2±1.0 gm/dl), which was found to be highly significant (p<0.001).

Prevalence and spectrum of von Willebrand disease in Eastern Uttar Pradesh.

Context: Von Willebrand disease is the most common inherited bleeding disorder with a prevalence of ≈ 1% in the general population. Studies available from India are limited, showing a prevalence of ≈ 10% of vWD amongst inherited bleeding disorders. Aims: This study aims to know the prevalence and subtypes of vWD in patients presenting with various bleeding manifestations to university hospital. Settings and Design: We investigated 840 patients presenting with bleeding manifestations in the period from August 2004 to August 2008 for bleeding disorders. Materials and Methods: Tests performed for the diagnosis of vWD included platelet count, bleeding time (BT), prothrombin time (PT), activated partial thromboplastin time (APTT), Factor VIII:C assay, von Willebrand Factor Antigen assay and Ristocetin-induced platelet aggregation. Results: Amongst 840 patients, 230 (27.38%) had inherited bleeding disorder. Out of these, 40 (17.39%) patients were identified as vWD. Type 1 in 17 (42.5%), Type 2 in 11 (27.5%) and Type 3 vWD was present in 12 (30.0%) patients. Patients' age ranged from five months to 45 years, with 17 males and 23 females. Positive family history was seen in 12 patients (30%). Muco-cutaneous bleeding was the most common presentation. Menorrhagia was present in 100% women of reproductive age group, and hemarthrosis was seen in two male patients. Conclusions: We felt that Type I vWD with infrequent and mild bleeding episodes remain undiagnosed either because of unawareness of the disease in society or due to paucity of diagnostic facilities available in our country. Therefore, an awareness program along with enhancement of diagnostic facilities for vWD is required in our country to identify these patients for proper management thus avoiding hemorrhagic complications.

Unusual presentations of visceral leishmaniasis.

We report 3 cases of Visceral leishmaniasis, who presented with unusual clinical manifestations. One child was diagnosed as hemophagocytic syndrome; another masqueraded with features of leukemoid changes alongwith hemophagocytosis and trilineage myelodysplasia; the third case presented with pyothorax. All the three patients showed amastigote forms of Leishmania donovani and positive serology (rk39 antigen). They showed complete clinical, hematological and parasitological resolution with Amphotericin B therapy.

Hemoglobin E disorders in Eastern Uttar Pradesh.

The distribution of hemoglobin E (alpha2beta2 26Glu (R)Lys ) is mostly restricted to Northeastern India. While evaluating the patients of jaundice, we came across two cases of hemoglobin E (Hb E) disorders. The first case is in a 22-year-old Bengali male and the second case of Hb E/beta thalassemia in a 5-year-old Hindu boy. The family study revealed Hb E trait in both the parents of Case 1, whereas in Case 2, the father was found to have Hb E trait and the mother had beta -thalassemia minor, thus confirming the diagnosis. Herein, we present the laboratory diagnosis and comparative data of the spectrum of Hb E disorders (i.e., heterozygous Hb E trait, homozygous Hb E disease and compound heterozygous Hb E/beta -thalassemia) that was found in our index cases and their parents.

Simultaneous appearance of dual malignancies of hematopoietic system-multiple myeloma and acute myeloid leukemia.

We herein report a case of denovo and simultaneous appearance of multiple myeloma and acute myeloid leukemia in a 58-year-old female patient, without prior exposure to chemotherapy or radiotherapy. This case is reported because of its extreme rarity.

The incidence and clinical characteristics of the immune phase eye disease in treated cases of human leptospirosis.

BACKGROUND: Uveitis is increasingly being reported from south India following epidemics of leptospirosis. The incidence of eye involvement in treated patients has not been investigated properly in prospective studies. AIMS: To determine the incidence, clinical spectrum and risk factors for Leptospiral uveitis in antibiotic-treated patients. SETTINGS AND DESIGN: A prospective cohort study conducted among the patients treated for leptospirosis at Calicut Medical College between July and November 2002 and a seroprevalence study among sewage workers. Materials And Methods: As many as 360 patients admitted with suspected leptospirosis were studied by clinical examination, baseline ophthalmic survey, laboratory investigations and Leptospira serology (Microagglutination test - MAT; and IgM and IgG using ELISA) during the epidemic. Of the 282 seropositive and antibiotic-treated cases, 174 patients who completed 30 months of regular follow-up were analyzed. A cross-sectional serosurveillance also was performed among 50 sewage workers to determine the baseline MAT titer. Statistical Analysis: Univariate analysis and logistic regression. RESULTS: Thirty-two patients (18.4%) developed the eye disease during follow-up. The mean age was 43.9 years and the sex ratio was equal. Twenty-one patients (65.6%) had anterior uveitis. Only six patients (18.8%) had visual symptoms. Median duration for developing anterior uveitis was 4 weeks. Recurrent uveitis was not seen following treatment. None had vision-threatening eye disease. Clinical and laboratory abnormalities during the acute phase did not pose risk for development of the eye disease later. Forty-six sewage workers (92%) showed a MAT titer of 1/25. CONCLUSIONS: Uveitis is common following acute leptospirosis. Antibiotic-treated patients during the acute phase of illness developed only mild uveitis.

Spectrum of anemia in pregnant Indian women and importance of antenatal screening.

Anemia is a common health problem but control of anemia in pregnant women is less well studied. The purpose was to study prevalence of anemia in young pregnant women, correlate with indices and study significance of identification of hemoglobinopathies. Of the 120 pregnant women, Hb was less than 8 g% in 58 (44.2%). Seventy-eight (65%) had iron deficiency, 22 (18.3%) had dimorphic anemia, and 14 (11.6%) had hemolytic anemia. Megaloblastic anemia was present in 6 (5%). Of hemolytic anemia, 50% were thalassemia trait. MCV< 76 fl was observed in 88 (73.3 %) cases. MCV<76 fl and MCH < 27 pg had 100 % sensitivity and 28.7 % specificity for screening of beta-thalassemia trait. NESTROFT had comparable sensitivity but lower specificity (14.9%). Sixty-three percent (60/78) of IDA had increased RDW whereas 78 % (11/14) of hemolytic anemia had RDW value in normal range (p value< 0.05). MCV/RBC of <14 was more specific parameter (96.8%) for beta-thalassemia trait. Four high-risk couples were identified. Thus, moderate to severe anemia was observed in most pregnant women. Hemoglobinopathies should be screened in antenatal clinics to identify the couples that would need a prenatal test. A lower MCV/RBC with RDWin the normal range may be useful in screening for thalassemia trait in pregnant women.

Risk factors for mortality in patients with leptospirosis during an epidemic in northern Kerala.

BACKGROUND: Epidemic leptospirosis is increasingly being reported from northern Kerala during the monsoon months. We investigated the risk factors for mortality during the 2002 epidemic. METHODS: Three hundred and forty patients suspected to have leptospirosis during the epidemic were studied by clinical examination, laboratory investigations and Leptospira serology (microscopic agglutination test). Two hundred and eighty-two seropositive cases were analysed for the clinical and laboratory profile, and risk factors for mortality using univariate and logistic regression analysis. RESULTS: Of the 282 seropositive cases, 58.9% were men. No significant association with occupational risk factors was seen; 62.9% had wounds on the feet. The majority had Weil syndrome with hepatic (69.8%) and renal (56.3%) involvement. Thrombocytopenia (65.8%) was common. Transient hyperglycaemia was observed in 10.3% of cases. Pulmonary haemorrhage (4.7%) and meningism (4.3%) were less common. Jaundice occurred in 46% of cases in the first week. The mortality rate was 6.03%. Hyperkalaemia (OR= 27.3), meningism (OR= 10.6), oliguria (OR=8.2), haemoptysis (OR= 5.4), bilirubin > 15 mg/dl (OR= 5.4), disorientation (OR=5), tachycardia (OR=4.1) and muscle tenderness (p=0.03) were the predictors of high mortality in univariate analysis. Only involvement of the lung and central nervous system were significant predictors of death in logistic regression. CONCLUSIONS: Leptospirosis is no more a mere occupational hazard in Kerala. Early occurrence of complications such as hepatitis mandates caution in the primary care setting. Lung and central nervous system involvement are significant predictors of mortality.

Dna probes for identification of leptospires and disease diagnosis.

A newly identified 1 kb DNA fragment amplified by PCR using (AG)8T inter-simple sequence repeats (ISSR) primer and a 631 bp segment of 16S rRNA ribosomal gene amplified by PCR using reported primers were labeled with a alpha32P dCTP for use as DNA probes. These probes were hybridized with DNA extracted from 19 standard pathogenic serovars, 3 standard saprophytic serovars, 33 pathogenic isolates (12 from patients, 1 from a tapwater source, and 20 from rodents), and 22 saprophytic isolates from environmental sources. The pathogen-specific 16S rRNA DNA probe specifically hybridized all 33 standard pathogenic serovars, to 13 pathogenic isolates. Similarly, the saprophyte specific 1 kb ISSR DNA probe specifically hybridized the 3 standard saprophytic serovars and the 22 saprophytic Leptospira isolates. The sensitivity of the 1 kb labeled saprophytic Leptospira specific DNA probe was 1.95 ng, and for the 16S rRNA pathogen specific probe 3.90 ng. The 16S rRNA gene segment DNA probe could also identify the leptospiremic stage in mice or guinea pigs infected experimentally with the pathogenic serovars australis, autumnalis or icterohaemorrhagiae. DNA probes therefore, owing to their high specificity and sensitivity, appear useful for easy, rapid, and reliable differentiation of pathogenic Leptospira strains and also hold promise for direct identification of organisms in blood samples to diagnose leptopsirosis.

Characterization of outer membrane proteins of Yersinia pestis and Yersinia pseudotuberculosis strains isolated from India.

The majority of virulence factors including the 12 Yersinia outer membrane proteins (Yops), 29 Yop secretion proteins (Ysc) and few specific Yop chaperone (Syc) are contributed by the 70 kb LCR middle plasmid of Yersinia pestis. Yersinia pestis isolates recovered during 1994 plague outbreak and rodent surveillance samples of Southern states of India were studied for the presence of important Yops by the conventional procedure of partially purifying outer membrane proteins (Omps) after cultivation in calcium deficient media. Prominent bands numbering 4-5 between 34-42 kDa region corresponding to important Yops were seen in all the isolates as well as in other Yersinia and non-Yersinia species by SDS-PAGE. Western blotting with the polyclonal antisera raised against these Omp preparations revealed few immuno-reactive bands that appeared to be shared among Y. pestis, Y. pseudotruberculosis, Y. enterocolitica, Y. fredrocksenii, Y. intermedia, Y. kristensenii and E. coli. Three recombinant Yop proteins namely, YopM, YopB and LcrV were produced and antisera to these proteins could reveal presence of these Yops in the Y. pestis Omp preparations. In order to further characterize the important Yops among Omps, attempts were made to generate monoclonal antibodies against Omp preparation. Three of the 4 stable reactive clones that were obtained, when tested, had extensive cross-reactions among pathogenic Yersinia species, Y. pestis and Y. pseudotuberculosis isolates, other Yersinia species and the members of Enterobacteriaceae in dot-ELISA and Western blotting. One of the monoclonal antibodies, YP1, exhibited reaction to all the pathogenic Yersinia species and the isolates, with restricted cross-reactivity to Y. intermedia, Y. kristensenii, K. pneumoniae. None of the 4 monoclonal antibodies had reactions with the 3 recombinant Yop proteins. It appears that under low calcium response, the Y. pestis not only activates secretion of Yops but also a large number of other proteins, which as per the present observations are cross-reactive within the family Enterobacteriaceae.

Hereditary coagulation factor X deficiency--a clinico-haematological profile of 5 cases from Varanasi.

Hereditary factor X deficiency is one of the rarest inherited coagulation factor deficiency. We are presenting here clinico-haematological profile and diagnostic approach of five such cases seen by us over a period of eleven years (year 1991-2001).

Acute megakaryoblastic leukaemia: a clinico-haematological profile of five cases.

Herein we are presenting the clinical, morphological and cytochemical characteristics of five cases of acute megakaryoblastic leukaemia (AML-M7) seen by us over a period of five years (Jan 1996-Dec 2000). Morphological assessment revealed marked polymorphism of blast cells and platelets both in the peripheral blood and bone marrow smears in all cases. Size of the blast cells ranged from very small to very large multinucleated cells, with variable chromatin pattern and number of nucleoli. More differentiated megakaryocytic cells showing cytoplasmic blebs, protrusions and platelet budding with bizarre platelet morphology were characteristic features suggesting the diagnosis. Cytochemical stains like myeloperoxidase, sudan black and PAS were positive in 5-15% of blast cell. Coagulation studies revealed a normal coagulation profile, whereas platelet studies showed marked impairment in aggregation of platelets with ADP and adrenalin with a normal PF-3 availability.

Use of restriction endonucleases for differentiation of pathogenic & saprophytic leptospires.

BACKGROUND & OBJECTIVES: PCR has been reported for amplification of the 482 bp genus specific region in 23S rRNA gene of Leptospira species. The sequence of this region was analyzed for specific restriction sites that could have yielded digestion products expected to provide differentiating banding profile for the pathogenic and the saprophytic group of leptospires. METHODS: Sixteen standard serovars of pathogenic group, two standard serovars of saprophytic group, 12 Leptospira isolates recovered from hospitalized patients with fever and jaundice or pyrexia of unknown origin and 23 isolates from different water sources were studied. Conventional tests, PCR methods and restriction digestion were used for confirming the identity of these isolates. RESULTS: All 12 isolates from patients and 1 from tap water source were identified as pathogenic and 22 isolates from water sources as saprophytic by the conventional tests and PCR. Of the 5 restriction endonuclease enzymes, viz, Apa I, Ban II, Hae III, Pst I and Sin I analyzed for digestion of PCR amplified 482 bp product, Apa I, Ban II, Pst I and Sin I provided fragments of different sizes providing distinct patterns for saprophytic and pathogenic leptospires. INTERPRETATION & CONCLUSION: The identity of a strain to genus Leptospira could be confirmed by PCR amplification of 482 bp region of 23S rRNA and with further restriction digestion a clear distinction into pathogenic or saprophytic group was achieved with the use of any of these 4 restriction enzymes.

Pure red cell aplasia associated with thymoma.

A case of thymoma with mixed spindle and lymphocytic variety along with acquired, secondary, chronic pure red cell aplasia is an uncommon entity. The presented case is a case of anterior mediastinal tumour with marked anemia. On histopathologic and hematological examinations, it proved to be a case of thymoma along with pure red cell aplasia. High index of suspicion, bone marrow examination, radiological including CT scan examination, total thymectomy after preparation with repeated blood transfusion remains the mainstay of treatment. Prolonged corticosteroid therapy leads to remission for upto ten months after operative intervention.

Chronic lymphocytic leukaemia terminating into plasma cell leukaemia.

Transformation of chronic lymphocytic lymphoma into plasma cell leukaemia is extremely rare. The diagnosis is made on if the circulating plasma cells in peripheral blood is in excess of 2000 cells/mm3.

Hereditary combined coagulation factor V and factor VIII deficiency: report of two Indian families from Varanasi.

Hereditary deficiencies of blood coagulation factors usually involve a single protein defect. Herewith we are describing clinical features and laboratory approach for the diagnosis of combined coagulation factor V/VIII deficiency which we encountered in 3 patients from 2 unrelated Hindu families of Varanasi.