Splenomegaly, cardiomegaly, and osteoporosis in a child with Gaucher disease.

A 15-month-old girl, born to the consanguineous parents, was referred with the sign of massive splenomegaly associated with thrombocytopenia and anemia. Plasma Chitotriosidase estimation was carried out as a screening test and was found to be normal with reduced activity of ß-glucosidase in leucocytes suggestive of Gaucher disease. At the age of 4 years, severe osteoporosis and cardiomegaly with pulmonary congestion were observed in the child. Molecular analysis for GBA gene has revealed homozygous status for L444P (c.1448C) in the proband, whereas parents and two elder sisters were found to be heterozygote. Prenatal study during the fourth pregnancy was carried out from cultured chorionic villi for ß-glucosidase, which was in the carrier range. Further confirmation of the carrier status was carried out from amniotic fluid DNA and was found to be heterozygous for L444P (c.1448C) in the GBA gene. This case demonstrates that children with the sign of splenomegaly with anemia and thrombocytopenia need to be screened for Gaucher disease, and molecular study can further help to confirm the heterozygous status, where prenatal study by enzyme investigation demonstrate heterozygous condition.

Beta-thalassemia mutations in western India.

OBJECTIVE: To study occurrence of common mutations in the population of Gujarat and the most prevalent mutation in certain high-risk communities. METHODS: The mutation screening was carried out using ARMS-PCR in children with beta thalassemia. RESULTS: Population screening has identified certain communities like Sindhis, Lohana, Rajputs, and SC/ST/OBC to be at higher risk as compared to others. The most common mutation was IVS 1-5 (G-->C) followed by 619 bp deletions of the total cases coming to Gujarat. CONCLUSION: Molecular evaluation for Thalassemia should be considered for families whose ethnicity indicates origin from high-risk community.

Cytogenetics and fluorescence in-situ hybridization in detection of hematological malignancies.

BACKGROUND: The technique of Fluorescence In-Situ Hybridization (FISH), a hybrid of cytogenetics and molecular biology has increased the resolution and application of cytogenetics in various neoplastic processes. In various types of leukemias, primary investigation by conventional cytogenetic [CC] technique followed by FISH has increased our understanding of the abnormal clonal formation involving different gene region. AIMS: Present study is aimed to use different kinds of in-house FISH probes in various hematological malignancies and its correlation with conventional cytogenetic finding. MATERIAL AND METHODS: Cytogenetic study was carried out in 360 patients either from peripheral blood or from bone marrow cells suspected for various types of leukemias. Four of 360 cases were further selected for FISH study by using different types of in-house probes, such as BAC [Bacterial Artificial Chromosome], PAC [Phague Artificial Chromosome], alphoid, PCP [Partial Chromosome Paint] and WCP [Whole Chromosome paint]. RESULTS: The results confirmed breakpoints of inversion 16 and del 16 in case 2 and 3 respectively. Whereas, case 1 did not confirm the cytogenetic findings of t(15;17) by PML/RARa fusion signals as multiple cell lines were involved in the patients. PCP and WCP were helpful in the identification of the marker chromosome in case 1. Telomeric and centromeric probes confirmed the cytogenetic findings of t(5;7) in case 4. CONCLUSION: We observe from this study that, in addition to the conventional cytogenetic study, FISH study provide further confirmation of chromosomal rearrangements. This facilitates our understanding of the neoplastic process more precisely for the better prognostication of the patient.

Usefulness of cytogenetics in leukemias.

Present study consists of cytogenetic evaluation in 141 cases referred to our centre for various leukemias. This includes 110 cases of CML, 10 of ALL, 16 of AML (M3), 2 of AML(M2), 2 of MDS and 1 of CMML. The conventional cytogenetic study was carried out in all the cases using G Banding technique. Of the 141 patients studied, 17 patients showed secondary chromosomal alterations along with primary chromosomal alterations. In two patients of CML with secondary chromosomal alteration t(4:9:22), molecular cytogenetic technique (FISH) has been carried out which has confirmed the primary observations revealed by the conventional cytogenetic technique. Other secondary alterations were numerous and would have been missed if only FISH or PCR technique would have been used for diagnosis. We observed from our study that advanced molecular techniques like FISH and PCR cannot replace the conventional cytogenetic study but are useful as supportive and confirmative diagnostic tools.

Study of anticardiolipin antibodies in repeated abortions--an institutional experience.

A cohort of 178 pregnant women with a history of first or second trimester abortions (2 or more) were the base of present study. In all, other causes of abortion were ruled out except for anti-phospholipid syndrome. Anti-Cardiolipin antibody (ACA) (IgG & IgM) was estimated in the sera samples of all women. Out of 178 women, any one or both immunoglobulins were above the cut off range (> 15.0 units) in 47 (26.4%) while both immunoglobulins were normal in 131 (73.59%) women. Both immunoglobulins were present in only 0.5% women. ACA-IgG alone was present in 11.79% while ACA-IgM alone was present in 14.04% women. We observe from present study that ACA is a major cause of recurrent fetal loss & many pregnancies can be saved if diagnosed & treated adequately.

Diabetes, microalbuminuria and hypertension.

Diabetes is a chronic condition which poses a risk for three major complications. They are diabetic retinopathy, nephropathy and neuropathy. Almost one third of diabetic patients (IDDM or NIDDM) develop diabetic nephropathy in their life time. Because of increased vascular permeability in chronic conditions increased urinary albumin excretion in the range of 30-200 mg/L (microalbuminuria) gives an early signal of incipient diabetic nephropathy. The prevalence of microalbuminuria was found to be 41% in diabetic patients with duration of more than 5 years. Seventy percent of diabetic patients with microalbuminuria were hypertensive. ACE inhibitors are shown to have significant effects on microalbuminuria and hypertension. We conclude that microalbuminuria is an early feature of excessive capillary leakage and its assessment in diabetic patients with duration of more than 5 years provides a simple non-invasive method of early diagnosis of incipient diabetic nephropathy. An early intervention may retard the progression to end-stage renal disease (ESRD).

Sub-biochemical hypothyroidism: an exaggerated thyroid stimulating hormone response to thyrotrophin releasing hormone.

BACKGROUND: The availability of sensitive and specific assays for evaluation of the thyroid axis has allowed definition of thyroid disorders at subclinical stage. This has almost obviated the use of thyrothrophin releasing hormone (TRH) study. We describe here a group of patients with minimal signs of hypothyroidism having normal thyroid function tests (T3, T4, thyroid stimulating hormone (TSH)) and have shown exaggerated TSH response to TRH. MATERIAL AND METHODS: Total 82 subjects were studied. Of these, 11 were age and sex matched controls, and 71 were patients. In all subjects TSH and other thyroid assays (T3, T4, FT4) were done by immunoradiometric assay (IRMA), and radioimmunoassay (RIA) respectively. Thyroid antibody was carried out by haemagglutination method. Results were compared to age and sex related normal ranges. To further investigate the status of thyroid axis, TRH study was carried out using standard protocol. RESULTS: Based on TRH study patients were grouped in three categories. Group 1 included 29 patients whose TSH response to TRH was normal. Group 2 included 20 patients with normal baseline TSH and exaggerated TSH response to TRH and Group 3 included 18 patients with baseline TSH in the range of 5 to 10 mu IU/ml and exaggerated TSH response to TRH. There was a significant difference to total T3 between group 1 and 3 (p < 0.05) but mean values were within normal limits. While no significant difference was observed in total T4 between controls and patient's group. Serum TSH values were high in group 3 as compared to controls and Group 1 and 2 (p < 0.0001). For Free T4 no statistical significance was observed between Group 1, 2 and 3. Thyroid antibodies were positive in 22.7% of patients in Group 2 and 33.33% in Group 3. CONCLUSION: We conclude from the present study that even with sensitive TSH assays TRH study still has a role to mark the early stage of hypothyroidism. Those with a normal or upper normal TSH with exaggerated response to TRH are termed as sub-biochemical hypothyroidism and can be considered for thyroid replacement therapy.

The gene for autosomal dominant hidrotic ectodermal dysplasia (Clouston syndrome) in a large Indian family maps to the 13q11-q12.1 pericentromeric region.

Hidrotic ectodermal dysplasia (HED), Clouston syndrome (MIM No. 129500), is an autosomal dominant disorder affecting the skin and its derivatives. It is characterized by alopecia, dysplastic nails in hands and feet, and hyperkeratosis of the palms and soles. We have studied a large Indian pedigree (UR005), from Gujarat region, consisting of a total 127 individuals including 41 affected (12 males and 29 females). The phenotype in this family ranged from atrichosis to hypotrichosis, sparsity or absence of eyebrows, and thickening of palms and soles. In order to map the disease locus by linkage analysis, DNA polymorphisms were used in DNAs from 23 affected and 8 normal individuals. While genotyping was in progress, Kibar et al. [1996] reported mapping of the locus of a similar disease in French-Canadian families to 13q around marker D13S141. We then utilized markers on 13q to genotype the members of the Indian family. Linkage with 13q11-12.1 markers was confirmed with a maximum lod score of 3.27 (theta=0.00) with locus D13S1316. Multipoint linkage analysis yielded a lod score of 5.04 at theta=0.00 with D13S1316; haplotype analysis indicated that the gene for the Clouston syndrome in this family is localized proximal to D13S292. These data suggest that the gene for the Clouston syndrome in this Indian pedigree is probably the same as that described in the French Canadian families. The combination of data from all available families linked to 13q11-12.1 will make it possible to narrow the critical region and facilitate the positional cloning of the elusive gene.

Incomplete gonadal dysgenesis.

A 21-year-old phenotypic male with ambiguous external genitalia and hypospadias was referred for cytogenetic studies. Exploratory laparotomy revealed presence of a small atropic uterus, unilateral gonadal dysgenesis with fallopian tubes on both sides and a cryptorchid testis on the left side. No gonad could be traced on the right side. Chromosomal analysis from peripheral whole blood culture revealed a 46,XY cell line. No mosaicism was detected. Endocrine studies showed elevated levels of serum FSH and LH with low borderline testosterone level and failure to respond to HCG stimulation. The presence of fallopian tube next to testis suggests absence of anti-Mullerian hormone secretion by Sertoli cells. The absence of Wolffian duct derivatives indicates insufficient secretion of testosterone by Leydig cells.

Investigation into the possible mechanisms involved in altered digoxin levels in diabetic patients.

The present study was undertaken to investigate the possible factors which may contribute to the altered digoxin levels in diabetic patients. The digoxin levels were found to be significantly higher in diabetics (1.74 +/- 0.09 ng/ml) as compared to non-diabetics (0.76 +/- 0.07 ng /ml). There was a positive correlation between digoxin levels and glycosylated haemoglobin levels. All diabetic patients had serum creatinine, urea and potassium levels within normal limits. However, serum TSH levels were found to be significantly higher in diabetics as compared to controls. Serum tri-iodo-1-thyronine (T3) levels were found to be lower in diabetics as compared to non-diabetics. Our data suggests that diabetes-mellitus causes alteration of digoxin levels. One of the causes of this increase in digoxin levels may be a tendency towards mild hypothyroidism associated with diabetes mellitus.

Load transfer of posts and cores to roots through cements.

This study evaluated differences in load transfer when cast posts are fixed to roots with different cements. Cast posts and cores were cemented with 40 endodontically prepared teeth by using four different cement mediums. The load exerted on the root surface through the post was evaluated using a strain gauge. The increased flexure after cementation was statistically analyzed. In conclusion, there was load transfer from post to root structure when posts were cemented, but no difference was found between cementation mediums.

Etched-porcelain resin-bonded full-veneer crowns: in vitro fracture resistance.

A recent study showed that both anterior and posterior glass ionomer-bonded crowns were significantly weaker than natural teeth. However, the fracture resistance of resin-bonded anterior crowns exceeded the strength of their size-matched controls. The three cement types were significantly different, with the average strength for Gluma, a-bonded crowns being the highest, then the Scotch-bond, b-bonded crowns, while Ketac Bond, c glass ionomer cemented crowns were the weakest. This pilot study indicates that the resin-bonded etched-porcelain full-veneer crowns are almost as strong as natural teeth and significantly stronger than glass ionomer-bonded crowns.

Effect of etching glass-ionomer cements on bond strength to composite resin.

The use of glass-ionomer cement in restorative dentistry has seen a revival because of its capacity for being etched and bonded to composite resin. Past investigators compared an etched cement surface with an unetched surface that was set against a smooth surface. Clinically, however, a glassy smooth surface is not produced when the cement is used as a base. Using Scotchbond bonding resin, we developed this two-part study to evaluate the tensile bond strengths of P-30TM composite resin to several glass-ionomer cements that were (a) unetched but allowed to set in air and (b) etched for 30 s with orthophosphoric acid, and to compare them with the cohesive strength of the respective cement. Using a silver nitrate staining technique, we also evaluated the microleakage of class V cavities restored with SiluxTM composite resin under a base of etched or unetched Ketac Bond cement. Although there were significant differences among three cements between their cohesive strength and the resin bond strength after the two surface treatments (p less than 0.01), the bond to the unetched surface was generally similar to that of the etched surface of the cement. The remaining groups showed no statistical difference. The microleakage was similar in the two groups. SEM micrographs showed a rough topography of the unetched cement that resembled that of the etched surface. This in vitro study suggests that acid-etching a glass-ionomer base for resin-bonding may not be necessary for specific materials. Further clinical evaluation is recommended to validate this observation intra-orally.

Cuspal deformation and fracture resistance of teeth with dentin adhesives and composites.

This study investigated the effect of polymerization shrinkage of posterior composite (Herculite) on the dimensions and fracture strength of human maxillary premolars with a phosphonate-ester DBA (Bondlite) and a second DBA comprised of 4-META with MMA/TBB (Superbond). The individual role of etched enamel and nonetched dentin bonding was also studied with high-copper amalgam used as a control. Two sizes of preparation isthmus were designated for evaluation of the effect of the polymerization contraction on weakened teeth. The following conclusions were drawn. 1. Narrow intracoronal tooth preparations were significantly stronger than wider, expansive preparations. There was no difference among the narrow isthmus preparations restored with amalgam or acid etching and composite. 2. There was a significant expansion in tooth dimension when MOD cavities were restored with amalgam. 3. Teeth with narrow isthmus preparations using composite bonded with Bondlite adhesive to etched enamel demonstrated a significant contraction compared with Superbond DBA. 4. Composite restorations using Superbond DBA in wide MOD cavities significantly improved the fracture strength of maxillary premolars. 5. Superbond DBA with composite restorations benefited the tooth in dimensional change and fracture strength. 6. There was a positive correlation between the fracture strength and tooth dimension.