Subhadra Sharma

MTHFR (677 and 1298) and IL-6-174 G/C genes in pathogenesis of Alzheimer's and vascular dementia and their epistatic interaction

Genetic risk factors play an important role in the pathogenesis of Alzheimer disease (AD) and vascular dementia (VaD). In this case-control study, we examined C677T and A1298C (rs1801133 and rs1801131) polymorphism in the methylenetetrahydrofolate reductase (MTHFR) genes and their correlation with plasma levels of homocysteine (Hcy) in AD and VaD cases and evaluated the gene-gene interaction (epistasis) with IL-6-174 G/C (rs1800795). CC genotype was associated with elevated levels of plasma homocysteine (p = 0.004) as compared with genotype AA of rs1801131. In AD, we observed a significant (p = 0.04) association with C alleles of rs1801131. Regression analysis revealed that the presence of both rs1801133 T and rs1800795 C alleles increased the odds of developing AD by 2.5 and VaD by 3.7-fold. While rs1800795 (CC or GC) genotypes alone increased the odds of developing VaD by 2.2-fold, the presence of CC genotype of rs1801131 nullified this effect. The findings support the hypothesis that multiple genes are involved to alter the odds of developing AD and VaD.

Coagulopathy as prognostic marker in acute traumatic brain injury

Context: Coagulopathy frequently occurs following traumatic brain injury (TBI) and usually occurs 6-72 hour post-trauma. The incidence and the probable risk factors for development of coagulopathy and poor outcome following TBI are largely unknown and vary considerably. Aims: To assess the incidence and probable risk factors for development of coagulopathy and to identify the risk factors for poor outcome in terms of median survival time following TBI. Materials and Methods: In this prospective study over two years, patients of isolated moderate and severe traumatic brain injury (GCS≤12) admitted to trauma center had coagulation profile (PT, APTT, thrombin time, fibrinogen and D-dimer), arterial lactate and ABG analysis done on day of admission and on day three. Coagulopathy was defined as prothrombin time (PT) or/and activated partial thromboplastin time (APTT) more than 1.5 times the normal control. Incidence of in-hospital mortality was assessed in all cases. Statistical Analysis: A stepwise logistic regression analysis was performed to identify risk factors for coagulopathy and mortality in these patients. Results: A total of 208 patients were enrolled in the study. The mean age was 32 ± 12 years and mean GCS was 7.1 ± 2.8. Coagulopathy was present in 46% (n = 96) of patients. Risk factors for development of coagulopathy were found out to be severity of head injury (OR: 2.81), elevated D-dimer (OR: 3.43), low hemoglobin (OR: 3.13), and effaced cisterns in the CT scan (OR: 2.72). Presence of coagulopathy (OR: 2.97) and severity of head injury (OR: 5.70) strongly predicted poor outcome, and were associated with a decreased median survival time. Conclusions: There is a high incidence of coagulopathy following TBI. The presence of coagulopathy as well as of severity of TBI are strong predictors of in-hospital mortality in these patients.

Fetal liver infusion in aplastic anaemia

Forty patients with severe aplastic anaemia received an intravenous infusion of 0.004 to 11.1 x 10(8) (median: 8 x 10(8) hematopoietic cells prepared from the fetal livers of 8-32 week old abortuses. Five patients, who died within 15 days of fetal liver infusion, are excluded from analysis. Twenty-two of the 35 evaluable patients (62%) responded favourably. Six of the 7 patients with good response were alive after 9 to 44 months (median: m = 20); one died 106 months after fetal liver infusion due to renal lithiasis. Four of the 7 with moderate response were alive after 9 to 31 months; 3 died within 16 months. Of 8 patients with minimal response, one was lost to follow-up and the others died in 3.4 to 10 months (m = 6). Median survival of responders was 15.7 months. Bone marrow cellularity became normal in 12 patients following fetal liver infusion. In seven patients, there was a relapse; 6 regained a normal bone marrow cellularity after a second or third fetal liver infusion. These data strongly suggest a role of fetal liver infusion in inducing bone marrow recovery. Of 13 non-responders, 4 were lost to follow-up and 9 died within 20 days-4.3 months (m = 1.6). Fetal liver infusion appears to be an effective therapy in patients with severe aplastic anaemia.

Prognostic assessment of various parameters in chronic myeloid leukemia

In 152 cases of chronic myeloid leukemia (CML) (actuarial median survival [MS], 59.2 months), the statistical relation of individual parameters with survival was studied to ascertain their prognostic value. The following parameters were found to be unrelated to the survival: age, sex, duration of symptoms, sternal bone tenderness, degree of hepatomegaly, level of hemoglobin, and leukocyte and platelet counts at the time of diagnosis. Splenomegaly of less than 10 cm and duration of first remission of 6 months or more were associated with significantly longer survival (MS, 70.5 and 68.5 months, respectively) as compared to bigger spleen size and duration of remission of less than 6 months (MS, 50.5 and 26 months; P > 0.01 and P > 0.05, respectively). The most significant prognostic parameter was the time required to achieve first remission. MS was 70 months in patients who achieved first remission in 2 months or less; it was 23.5 months in the remaining patients. This difference was statistically highly significant (P > 0.001).

Serum cytokines and anxiety in adolescent depression patients: Gender effect

The present study compares the serum cytokine levels between adolescent depression patients and healthy controls and assesses correlation between depression, anxiety scores and serum levels of eight cytokines. Study also checked the variation in serum levels with medication status (medication free/naïve vs. patients on medication). Following clinical and psychometric assessment of 77 adolescent (aged 13-18 years) depression patients (49 males and 28 females; 56 medication free/naïve) and 54 healthy controls (25 males, 29 females), eight cytokines (IL-1β, IL-2, IL-6, IL-10, TNF-α, IFN-γ, TGF-β1 and IL-17A {denoted IL-17 throughout}) were measured in serum using ELISA. Depressed adolescents had significantly high levels of IL-2 (p<0.001) and IL-6 (p=0.03) as compared to controls. The female population skewed the result of one cytokine (IL-6) in patients. Anxiety scores showed positive correlation (only in female patients) with IL-1β, IL-10 and negative correlation with TGF-β1 and IL-17. The gender effect in relationship between anxiety and cytokines was not straightforward. On comparing study groups on the medication/naïve status, IL-2 and TGF-β1 showed significant difference between the groups (p<0.001, p=0.007 higher in medicated). Depression in adolescents was associated with elevation of proinflammatory serum cytokines with a gender bias for females. Anxiety scores correlated negatively with TGF-β1 and IL-17.

Synergistic Epistasis of Paraoxonase 1 (rs662 and rs85460) and Apolipoprotein E4 Genes in Pathogenesis of Alzheimer’s Disease and Vascular Dementia:

Genetic polymorphism and epistasis play a role in etiopathogenesis of Alzheimers disease (AD) and vascular dementia (VaD). In this case-control study, a total of 241 patients were included in the study to see the effect of paraoxonase 1 (PON1; rs662 and rs85460) and apolipoprotein E (ApoE) genes in altering the odds of having AD and VaD along with serum PON and lipid profile. The presence of at least 1 variant allele of rs662, but not rs85460, increased the risk of having AD by 1.8-fold (95% confidence interval [CI]: 0.97-3.40) and VaD by 3.09-fold (95% CI: 1.4-6.9). The interaction between PON1 genes (rs662 and rs85460) and ApoE genes showed synergistic epistasis in altering the odds of significantly having both AD and VaD. On the other hand, low serum level of high-density lipoprotein and low level of serum PON activity were found associated significantly (P ≤ .001 in both cases) only in patients with VaD as compared to healthy control.

Hemopoietic improvement following fetal liver infusion in aplastic anemia

Abstract: 41 (38 males, 3 females) patients with aplastic anemia received fetal liver infusion (FLI) from 74 abortuses with gestation periods of 8–32 (Median(M)‐16) weeks and cell dose of 0.004–11.1 times 108 (M‐2.3 times 108) from September, 1976 until November, 1987. 35 persons received single FLI; those with recurrence or no response received two or more FLI. 8 received two; 7, three; 2, four; and 1, six FLI. There was a slow and incomplete autologous hematopoietic improvement in 40% and expected survival of 52% at 1 year, 45% at 2 yr, and 37% at 5 yr (Kaplan Meier estimate). There was rise in fetal hemoglobin (Hb), 0–15.7%, (M‐3.5) among responders in 3–20 (M‐6) months. Patients who survived for more than 12 months had, on average, a longer duration of disease (4 months or more), and higher granulocyte and platelet counts. Statistically, however, these differences were not significant. Reticulocyte count was significantly lower in those who survived beyond 12 months. 1 patient developed acute undifferentiated leukemia 3 yr post‐FLI. The study indicates that fetal liver infusion is likely to benefit about 40% of individuals suffering from severe aplastic anemia. Longer surviving patients, however, may be at risk of developing clonal diseases.

Fetal liver infusion in acute myelogenous leukaemia

Forty-five patients with acute myelogenous leukaemia (AML) received induction chemotherapy with either a conventional dose of cytarabine and daunorubicin (27 patients) or a low dose of cytarabine (18 patients). Maintenance chemotherapy was given to all responders. In 14 of 39 evaluable patients, infusion of fetal liver cells from 10-24 weeks old foetuses was given following induction as well as maintenance therapy. Six of 14 patients (43%) achieved a complete remission; 2 showed evidence of transient engraftment documented by analysis of sex chromosomes and RBC antigens (1 patient each). Fetal liver infusion within 6 days of completing induction chemotherapy appeared more effective than when given later. Five of 25 patients (20%) who did not receive fetal liver infusion achieved a complete remission. The present work suggests that fetal liver infusion given following induction chemotherapy may increase the remission rate in AML either by temporary engraftment or by accelerating the rate of haematological recovery.

Haemopoietic cell composition of human fetal liver, spleen and thymus

The haemopoietic cell composition of fetal liver, spleen and thymus was studied in human aborted fetuses of 12–22 weeks gestation. Erythroid cells were present primarily in liver and to a lesser extent in spleen. Orthochromatic normoblasts were the predominant erythroid component. Lymphoid cells were seen primarily in the thymus and to a lesser extent in the spleen. There were few granulocytic and megakaryocytic cells in all these organs, A few primitive haemopoietic cells (Haemocytoblasts) were identified only in liver. These observations indicate that during mid-fetal life (12–22 weeks) the liver contributes primarily to erythropoiesis, the thymus to lymphopoiesis, the spleen to both, and that there is a lack of granulopoiesis and megakaryopoiesis at these sites.

Bone marrow recovery following fetal liver infusion (FLI) in aplastic anaemia: Morphological studies

Twenty-two of 35 patients with aplastic anaemia who received fetal liver infusion (FLI), responded to this treatment. Detailed review of bone marrow aspirates and biopsies was available in 17. There was a good correlation between clinical improvement and blood counts. The bone marrow cellularity increased to 48 percent in about 18 weeks with repopulation by both erythroid and myeloid cells. The erythroid response was predominant and the earliest to occur (1.5-61 weeks); it lasted for 7.5-100 weeks. Marked dyserythropoiesis was observed. Myeloid response occurred simultaneously in 45 percent of the responders; in the others it was delayed by 1.5 to 4 weeks. Significant dysmyelopoiesis with shift to the left, unrelated to infection, was seen 1.5 to 9.5 weeks after FLI lasting for 9-26 weeks in most cases. Megakaryocytic response either did not occur or was delayed, less marked and often transient.