Correlations between apoptotic and proliferative indices in cervical intraepithelial neoplasia and carcinoma

Aims of the present paper was to study cell death by apoptosis and cell proliferation in normal cervical biopsies, cervical intraepithelial neoplasms (CIN) and squamous cell carcinomas of cervix (CaCx). There were each seven cases of normal cervical biopsy, CIN1 and CIN2 along with 10 cases of CIN3 and 14 cases of CaCx. Percentage of apoptotic cells and bodies (i.e. apoptotic index, AI) and mitoses (i.e. mitotic index, MI) and turnover index (TI - AI + MI) were counted in formalin fixed, paraffin embedded, haematoxylin and eosin stained slides. AgNOR stain was done and mean AgNOR dots per cell was also estimated. AI, MI and TI were correlated with histology grade of CIN and invasive carcinoma cervix. Mean AI, MI, TI and AgNOR count increased from lower to higher grades of CIN. AI, MI & TI raised significantly from CIN3 to carcinomas; AI, TI & AgNOR count raised significantly from CIN1 to combined CIM2 & 3; TI & AgNOR count were high in CIN1 to CIN2; AI & TI were significantly raised in normal to CIN1. In conclusion, TI is probably more important for cell kinetic analysis of CIN and carcinoma of cervix because it reflects the frequency of two important events i.e. mitosis and cell death. Sudden increase of AI, MI, TI count from CIN3 to CaCx may indicate the possibility of genetic alteration of cells of CIN3 which induces a frank malignant transformation from CIN3 to CaCx.     

Genetic variation at three VNTR loci in three tribal populations of Orissa,India

BACKGROUND: The variable number of tandem repeats (VNTR) loci are robust, simple and rapid tools for genetic characterization of both individuals and populations. This paper presents data on the distribution of three VNTRs (APOB, YNZ22 and D1S80) in three tribal populations (Gadaba, Kuvi-Khond and Paroja) of the Koraput district of Orissa, India with a view to enlarge our understanding of molecular genetic diversity among these tribes and the usage of these VNTRs in anthropogenetic studies. SUBJECTS AND METHODS: Three tribal populations were genotyped for APOB, YNZ22 and D1S80 loci using the polymerase chain reaction (PCR) technique. Gadaba are an Austro-Asiatic tribe while Kuvi-Khond and Paroja are Dravidian tribes. All samples were collected, with consent, from unrelated individuals. RESULTS: The observed allelic variation in these tribes is comparable with many Indian populations, but they showed significant overall and inter-population variability within the region. Allele *24 was the most common allele at the D1S80 locus in all populations, with Gadabas having the highest frequency (50%) followed by Paroja (32%) and Kuvi-Khond (23%). Gadabas also showed a higher frequency of allele *19 (13%) and *31 (9%) compared to other Indian and European populations. In the Apo B system, allele *37 was the most common in all three populations, with Gadabas having the highest frequency (39%) followed by Paroja (24%) and Kuvi-Khond (21%). This allele is present in high frequencies in other Indian (except Gonds) and European populations. Alleles *33 (17%), *35 (20%) and *45 (12%) were also common in the Gadabas, but Kuvi-Khond showed higher frequencies of *31(10%), *35(13%) and the larger allele *49(16%). Paroja, on the other hand, had higher frequencies of *31 (14%), *33 (17%) and *45 (13%). Allele *49 was also present in Paroja (10%) but was absent in the Gadaba. For the YNZ22 system, allele *4 was the most common in Kuvi-Khond (25%) and Paroja (21.4%), and allele *2 was the predominant allele in the Gadaba (33%). However allele *4 still occurs at relatively high frequency in Gadaba (27%). Allele *2 also occurs at relatively high frequency in Kuvi-Khond (20%) and intermediate frequency in Paroja (11%). Average heterozygosity was relatively low for Gadaba (0.7597 +/- 0.0191) and high for Kuvi-Khond (0.8618 +/- 0.0149) and Paroja (0.8512 +/- 0.0190), perhaps a reflection of effective population size and limitations to mating. The level of gene differentiation is, however, low (3-4%) for the three systems studied in these tribal populations and in data compiled from previous studies from the region. CONCLUSIONS: The VNTRs are polymorphic in the tribal populations studied and there is extensive allelic variation. Gadabas are isolated but Kuvi-Khond and Paroja show clear affinities with the Gonds, a major tribal group of Central India. Overall, allele frequency distribution, heterozygosity and genetic diversity analysis show that genetic diversity observed is socially, linguistically and geographically structured in this region.     

DAT1 3'-UTR 9R allele: preferential transmission in Indian children with attention deficit hyperactivity disorder.

Genetic alterations in the dopaminergic system are frequently observed in association with attention deficit hyperactivity disorder (ADHD) and a 40 bp variable number of tandem repeats (VNTR) in the 3'-untranslated region (3'-UTR) of the dopamine transporter gene (DAT1) has been investigated in different populations. Both significant association and lack of association with the10 repeat allele (10R) of DAT1 VNTR have been reported. Objective of the present investigation was to examine association of this polymorphism with ADHD in Indian children. Genotypic data obtained from ADHD probands (n = 79), their parents (n = 148) and control individuals (n = 153) were analyzed for haplotype-based haplotype relative risk analysis (HHRR), transmission disequilibrium test (TDT), and family-based association test (FBAT). HHRR analysis revealed significant (P = 0.009) transmission of shorter alleles (< or =9R). TDT analysis of informative ADHD families (n = 32) also exhibited highly significant transmission of the shorter alleles (P = 0.002). Further analysis by FBAT showed preferential transmission (P = 0.019) of the 9R allele from parents to ADHD probands. It can be inferred from the data obtained that the DAT1 3'-UTR 9R allele may confer risk of ADHD in the Indian population.     

Thioredoxin regulation of ischemic preconditioning

Thioredoxins are a class of small redox-regulating proteins that appear to play a crucial role in many oxidative stress-inducible degenerative diseases. A recent study demonstrated a reduction of thioredoxin-1 (Trx1) protein in the ischemic reperfused myocardium. When the same heart was adapted to ischemic stress by preconditioning with repeated cyclic episodes of small duration of ischemia and reperfusion, there was an increased induction of Trx1 expression. Inhibition of Trx1 expression resulted in reduced postischemic ventricular recovery and increased myocardial infarct size in the preconditioned heart. Corroborating these findings, transgenic mouse hearts overexpressing Trx1 were resistant to ischemic reperfusion injury as compared with the hearts from wild-type mice. Thus, it appears that thioredoxin plays a crucial role in cardioprotection induced by preconditioning.     

Targeting myeloperoxidase to azurophilic granules in HL-60 cells

Myeloperoxidase (MPO) is a cationic protein and one of the major constituents of azurophilic granules in neutrophils. Here, we examined whether intracellular transport of MPO and serglycin, a chondroitin sulfate (CS)-bearing proteoglycan, is correlated. First, we examined binding of MPO to CS-Sepharose and measured an ionic interaction, which was disrupted by 200-400 mM NaCl. Next, HL-60 promyelocytes were activated with a phorbol ester, which induced an almost complete rerouting of serglycin from the granular to the secretory pathway, concomitant with a similar effect on MPO transport and secretion. We then used the membrane-permeable cross-linker dithiobis(succininmidylpropionate; DSP) after labeling HL-60 cells with [35S]methionine and [35S]cysteine for 19 h. Immunoprecipitation of MPO revealed its cross-linking to high molecular material having the appearance of a proteoglycan in sodium dodecyl sulfate-polyacrylamide gels. This assumption was confirmed by labeling HL-60 cells with [35S]sulfate for 10 min followed by DSP cross-linking and immunoprecipitation. From three granular enzymes immunoprecipitated, only the cationic MPO was cross-linked to [35S]sulfate-labeled serglycin in appreciable quantities, whereas cathepsin D or beta-N-acetylhexosaminidase was not. Thus, intracellular transport of MPO appears to be linked to that of serglycin. Extracts from high buoyant density organelles from human placenta containing MPO activity were subjected to CS-affinity chromatography. Proteins binding to CS were identified by mass spectrometry as MPO, lactoferrin, cathepsin G, and azurocidin/cationic antimicrobial protein of molecular weight 37 kDa, suggesting that serglycin may be a general transport vehicle for the cationic granular proteins of neutrophils.     

SPECT changes and their correlation with EEG changes in tuberculous meningitis

OBJECTIVE: In tuberculous meningitis (TBM) blood flow may be altered due to associated vasculitis, hydrocephalus and raised intracranial pressure. Electroencephalography (EEG) and single photon emission computed tomography (SPECT) provide information about electrical activity and regional cerebral blood flow respectively. This study aims at the correlation of EEG and SPECT changes in patients with TBM. METHOD: Sixteen patients with TBM whose age ranged between 5 and 62 years and 3 of whom were females were subjected to clinical, radiological (CT and/or MRI), EEG and SPECT studies using 99mTc ethylene cystine dimer (ECD). Ten patients were in stage III and 3 each in stage II and stage I meningitis. Cranial CT scan was carried out in 15 and MRI in 4 patients. Hydrocephalus was present in 9, infarction in 7 and tuberculoma in 5 patients. RESULTS: SPECT studies were abnormal in all except 2 patients revealing basal ganglionic hypoperfusion in 14 and focal cortical hypoperfusion in 9 patients. The EEG was abnormal in 11 patients which included delta slowing in 5, theta slowing in 6, frontal intermittent rhythmic delta activity (FIRDA) in 3 and epileptiform discharges in 2 patients. All the patients with abnormal EEG had abnormal SPECT study except 1. In 4 patients, EEG was normal although there was subcortical hypoperfusion on SPECT. In spite of high frequency of focal cortical hypoperfusion (9 patients), EEG revealed focal abnormality in 3 patients only. CONCLUSION: It can be concluded that the SPECT reveals more frequent abnormalities compared to EEG and CT scan. Cortical hypoperfusion with or without basal ganglia hypoperfusion is associated with FIRDA and diffuse delta slowing on EEG.     

Validity of transition-zone dosimetry at high atomic number interfaces in megavoltage photon beams

Measurement of dose or dose perturbation factors at high atomic number interfaces are usually performed with a thin-window parallel-plate ion chamber. In a transition region, under nonequilibrium conditions, accuracy of ion chamber readings for the dose measurements has often been questioned. This paper critically analyzes the factors (stopping power ratio and charge collection) for the dose measurements at interfaces. Monte Carlo simulations were performed to investigate the secondary electron spectrum produced by photon beams and to calculate the stopping power ratios at the point of measurement. The validity of dose measurements was studied for the photon beams in the range of Co-60 gamma rays to 24-MV x rays at bone and lead interfaces with polystyrene, using thermoluminescent dosimeters, extrapolation chamber and several types of commercially available parallel-plate ion chambers. It is observed that for energies greater than 10 MV most parallel-plate chambers can be used to measure dose accurately. At lower energies, however significant differences between measured doses with different detectors were noticed. It is suggested that at high-Z interfaces and lower energies, the dose measurements should be performed with ultrathin-window parallel-plate ion chambers or extrapolation chambers.     

Vitamin B12 deficiency neurological syndromes: a clinical,MRI and electrodiagnostic study

BACKGROUND: Vegetarianism is an important cause of vitamin B12 deficiency, especially in countries like India. We managed 17 patients with neurological syndrome due to vitamin B12 deficiency in a tertiary care referral teaching hospital which caters to relatively affluent population. AIM: To evaluate neurophysiological and MRI changes in patients presenting with vitamin B12 deficiency neurological syndrome and interpret these is the light of reported autopsy findings. SETTING: Tertiary care referral teaching hospital. METHODS: Patients with vitamin B12 deficiency neurological syndrome diagnosed by low serum vitamin B12 and/or megaloblastic bone marrow were subjected to clinical evaluation and spinal MRI. The neurophysiological tests included nerve conduction studies, tibial somatosensory evoked potential (SEP), motor evoked potential (MEP) and visual evoked potential (VEP) studies. The recovery was defined on the basis of 6 months Barthel Index score into complete, partial or poor. RESULTS: There were 17 patients with vitamin B12 deficiency neurological syndrome, 3 were females and 12 lactovegetarian. The clinical syndrome was that of myelopathy in 8, myeloneuropathy in 5, dementia myelopathy in 3 and neuropathy in 1 patient. All the patients had impaired joint position and vibration sensation in the lower limbs and 4 had in upper limbs as well. Lower limbs were spastic in 13 and upper limbs in 2 patients. Spinal MRI revealed T2 hyperintensity in cervicodorsal region in 6 and cord atrophy in 3 patients. Sural nerve conduction was abnormal in 8 and peroneal conduction in 5 patients. In one patient all sensory nerve conductions were unrecordable but motor conductions were normal. Tibial SEP was abnormal in 12 out of 15 and lower limb MEP in 8 out of 12 patients. P100 latency of VEP was prolonged in 7 out of 13 patients. Right to left asymmetry was present in tibial SEP in 4 and VEP in 2 patients. At 6 months followup 2 patients improved completely, 7 partially and 3 had poor recovery. Clinical recovery correlated with MEP but not with SEP or MRI changes. CONCLUSION: The evoked potential and MRI changes in vitamin B12 deficiency neurological syndrome are consistent with focal demyelination of white matter in spinal cord and optic nerve. Myelopathic presentation is commoner and SEP is more frequently abnormal. The outcome at 6 months correlated with MEP changes.