Association of vascular endothelial growth factor single nucleotide polymorphisms on the prognosis of breast cancer patients.

BACKGROUND: Vascular endothelial growth factor (VEGF), a major mediator of vascular permeability and angiogenesis, may play a pivotal role in mediating the development and progression of breast cancer. In the present study, we examined the genetic variations of the VEGF gene to assess its possible relation to breast cancer. MATERIALS AND METHODS: A total of 200 patients with histologically confirmed cases of breast cancer and 200 healthy women were genotyped for VEGF single nucleotide polymorphisms (405G > C and -1154G > A) by polymerase chain reaction-restriction fragment length polymorphism analysis. Pre-operative plasma VEGF levels were determined by enzyme-linked immunosorbent assay in 200 women with breast cancer and in 200 normal female controls. RESULTS: The genotype frequencies of the +405G > C, -1154G > A polymorphisms did not show a significant deviation from the Hardy-Weinberg expectation. The minor allele frequencies of the +405G > C and -1154G > A polymorphisms among cases and controls were 33.5% (C allele), 31.5% (A allele) and 35% (C allele), 34.5% (A allele) respectively. +405GG and -1154GG genotypes were associated with higher levels of VEGF among breast cancer cases and controls. Increased plasma VEGF levels were significantly associated with, clinical stage of the disease (P = 0.035). CONCLUSION: Although none of the polymorphisms were significantly associated with breast cancer, some of the VEGF genotypes may influence tumor growth through an altered expression of VEGF and tumor angiogenesis.

Depletion of primary cilia in articular chondrocytes results in reduced Gli3 repressor to activator ratio, increased Hedgehog signaling, and symptoms of early osteoarthritis.

OBJECTIVE: Primary cilia are present in almost every cell type including chondrocytes. Studies have shown that defects in primary cilia result in skeletal dysplasia. The purpose of this study was to understand how loss of primary cilia affects articular cartilage. DESIGN: Ift88 encodes a protein that is required for intraflagellar transport and formation of primary cilia. In this study, we used Col2aCre;Ift88(fl/fl) transgenic mice in which primary cilia were deleted in chondrocytes. Col2aCre;Ift88(fl/fl) articular cartilage was characterized by histological staining, real time RT-PCR, and microindentation. Hedgehog (Hh) signaling was measured by expression of Ptch1 and Gli1 mRNA. The levels of Gli3 proteins were determined by western blot. RESULTS: Col2aCre;Ift88(fl/fl) articular cartilage was thicker and had increased cell density, likely due to decreased apoptosis during cartilage remodeling. Mutant articular cartilage also showed increased expression of osteoarthritis (OA) markers including Mmp13, Adamts5, ColX, and Runx2. OA was also evident by reduced stiffness in mutant cartilage as measured by microindentation. Up-regulation of Hh signaling, which has been associated with OA, was present in mutant articular cartilage as measured by expression of Ptch1 and Gli1. Col2aCre;Ift88(fl/fl) cartilage also demonstrated reduced Gli3 repressor to activator ratio. CONCLUSION: Our results indicate that primary cilia are required for normal development and maintenance of articular cartilage. It was shown that primary cilia are required for processing full length Gli3 to the truncated repressor form. We propose that OA symptoms in Col2aCre;Ift88(fl/fl) cartilage are due to reduced Hh signal repression by Gli3.

Observations on the flow characteristics of blood flow in arteriovenous fistulae (experimental).

Hemodynamic play a very significant role in the pathophysiology of intracranial arteriovenous malformation. The surgical decisions are based on the understanding of the complexities of the flow. Quantification of the abnormal flow is difficult. The mathematical models provide limited information due to the simplicity of the design of these models. Flow of fluid in a tube is very sensitive to small changes in the diameter. We studied the flow characteristics of a fistula by introducing accurately machined acrylic fistulae between the femoral arteries and veins of dogs. The influences of systemic arterial pressure, diameter of the arterial feeders, volume of blood flow, velocity of flow and the diameter of the shunt on the flow of blood across the shunt were studied. Our experiments suggest that the flow characteristics of an arteriovenous fistulae are complex and are influenced by small changes in the diameters of the fistula and the feeding artery. Our model demonstrates the occurrence of the anomalous flow reduction in the fistula and steal phenomenon and is therefore a more realistic representation of the clinical situation. The design of a mathematical model should include the diameter of the fistula if it is intended to replicate the hemodynamic characteristics of an arteriovenous malformation more faithfully.

Methylenetetrahydrofolate reductase gene polymorphisms and risk of acute lymphoblastic leukemia in children.

INTRODUCTION: Methylenetetrahydrofolate reductase (MTHFR) is a critical enzyme in folate metabolism and is involved in DNA synthesis, DNA repair and DNA methylation. Genetic polymorphisms of this enzyme have been shown to impact several diseases, including cancer. Leukemias are malignancies arising from rapidly proliferating hematopoietic cells having great requirement of DNA synthesis. This case-control study was undertaken to analyze the association of the MTHFR gene polymorphisms 677 C"T and 1298 A"C and the risk of acute lymphoblastic leukemia in children. MATERIALS AND METHODS: Eighty-six patients aged below 15 years with a confirmed diagnosis of acute lymphoblastic leukemia (ALL) and 99 matched controls were taken for this study. Analysis of the polymorphisms was done using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: Frequency of MTHFR 677 CC and CT were 85.9% and 14.1% in the controls, and 84.9% and 15.1% in the cases. The 'T' allele frequency was 7% and 7.5% in cases and controls respectively. The frequency of MTHFR 1298 AA, AC, and CC were 28.3%, 55.6% and 16.1% for controls and 23.3%, 59.3% and 17.4% for cases respectively. The 'C' allele frequency for 1298 A-->C was 43.9% and 47% respectively for controls and cases. The odds ratio (OR) for C677T was 1.08 (95% CI 0.48-2.45, p = 0.851) and OR for A1298C was 1.29 (95% CI 0.65-2.29, p = 0.46) and OR for 1298 CC was 1.31 (95% CI 0.53-3.26, p = 0.56). The OR for the combined heterozygous status (677 CT and 1298 AC) was 1.94 (95% CI 0.58-6.52, p = 0.286). CONCLUSION: The prevalence of 'T' allele for 677 MTHFR polymorphism was low in the population studied. There was no association between MTHFR 677 C-->T and 1298 A-->C gene polymorphisms and risk of ALL, which may be due to the small sample size.

Immunohistochemical characterization of cancer antigen in uterine cancers.

The pattern of cancer antigen (CA-125) expression by immunohistochemistry (IHC) was investigated in malignant and nonneoplastic endometrium in endometrial carcinoma. Ninety cases of primary uterine carcinomas (65 endometrioid [EM] carcinoma, 15 serous papillary [SP] carcinoma, 6 carcinosarcomas [malignant mixed müllerian tumors], and 4 clear cell carcinoma [CC]) and adjacent atrophic and/or hyperplastic endometrium were analyzed by IHC for CA-125 expression. The percentage and intensity of luminal, apical, basal, and diffuse cytoplasmic immunostaining of epithelial cells were categorized on a scale of 0-4. The immunoreaction score (IRS score) was calculated and correlated with the grade and stage of carcinoma according to the histologic type. CA-125 expression (3-4/4) was localized in apical borders of grade 1 and grade 2 EM carcinoma and was weak or negative (0-1/4) in grade 3 EM. Mucinous differentiation in EM was associated with intense luminal and apical staining. Squamous areas and stroma showed no staining at all. SP carcinoma and endometrial intraepithelial carcinoma showed much higher mean IRS score than EM. In malignant mixed müllerian tumors (MMMT), the epithelial component stained as above according to the type of epithelial cell differentiation of the neoplastic cells. Benign proliferative glands showed moderate apical luminal, basal, and cytoplasmic staining. Intense diffuse staining was observed in atypical complex hyperplasia. Different patterns of CA-125 immunostaining were observed in normal, hyperplastic, and neoplastic endometrium. IRS score correlated with the grade but not with the stage of EM carcinoma. The intense different staining pattern of endometrium with atypical complex hyperplasia suggests that CA-125 may be a useful diagnostic aid.

Serum levels of bone alkaline phosphatase in breast and prostate cancers with bone metastasis.

BALP activity in the sera of metastatic patients of breast and prostate malignancy has increased significantly. Our studies with patients in India conform the earlier reports that BALP may have a useful complementary role in the early diagnosis of bone metastases.

Cloning and characterization of a eukaryotic pantothenate kinase gene (panK) from Aspergillus nidulans.

Pantothenate kinase (PanK) is the key regulatory enzyme in the CoA biosynthetic pathway. The PanK gene from Escherichia coli (coaA) has been previously cloned and the enzyme biochemically characterized; highly related genes exist in other prokaryotes. We isolated a PanK cDNA clone from the eukaryotic fungus Aspergillus nidulans by functional complementation of a temperature-sensitive E. coli PanK mutant. The cDNA clone allowed the isolation of the genomic clone and the characterization of the A. nidulans gene designated panK. The panK gene is located on chromosome 3 (linkage group III), is interrupted by three small introns, and is expressed constitutively. The amino acid sequence of A. nidulans PanK (aPanK) predicted a subunit size of 46.9 kDa and bore little resemblance to its bacterial counterpart, whereas a highly related protein was detected in the genome of Saccharomyces cerevisiae. In contrast to E. coli PanK (bPanK), which is regulated by CoA and to a lesser extent by its thioesters, aPanK activity was selectively and potently inhibited by acetyl-CoA. Acetyl-CoA inhibition of aPanK was competitive with respect to ATP. Thus, the eukaryotic PanK has a distinct primary structure and unique regulatory properties that clearly distinguish it from its prokaryotic counterpart.

Circulating levels in serum of total sialic acid, lipid-associated sialic acid, and fucose in precancerous lesion and cancer of the oral cavity.

Total sialic acid (TSA), lipid-associated sialic acid (LASA), and fucose were estimated in sera of 35 patients with precancerous lesion of the oral cavity, 68 patients with cancer of the oral cavity, and 25 age- and sex-matched non-chewers of both tobacco and betal nut and nonsmokers as controls. Significant elevation in the serum levels of TSA and LASA were observed in patients with the precancerous and cancer lesions when compared with the controls. Serum TSA levels were elevated significantly in patients with cancer when compared with those with precancerous lesions. Circulating TSA and LASA levels were found to reflect tumor burden and correlated well with stage of the disease. However, serum fucose levels did not show an increase corresponding to stage of the disease. The results suggests that combined evaluation of these markers may be useful in predicting early malignant change and also in assessing the spread and invasiveness of the disease in cancer of the oral cavity.

Serum vitamins' status in oral leucoplakias--a preliminary study.

Vitamins, such as A, beta carotene, C, E, B12 and folate, are the micronutrients with the strongest evidence of having a link to cancer prevention and control. Deficiency of these vitamins at the dietary, systemic or mucosal level will interact with tobacco use and increase the risk of oral precancerous lesions. The objective of this study was to (1) establish the baseline circulating levels of these vitamins in our normal population with and without tobacco use and (2) compare these levels with the values obtained in cases of oral leucoplakias. 50 normal controls with 25 each in chewers and non-chewers, matched for age and sex, were selected. 50 cases of oral leucoplakias (clinically detectable white patches) from the field constituted the study group. Simultaneous measurement of serum vitamin B12 and folate were carried out by radioassay. The other serum vitamins were estimated spectrophotometrically. Except for serum vitamin E, all the other serum vitamin levels were significantly decreased in oral leucoplakias compared to the controls. Cancer chemopreventive agents acting as inhibitors of both initiation and promotion, as analysed in our population, is promising for further intervention trials.

Serum carotene, vitamin A, and vitamin C levels in breast cancer and cancer of the uterine cervix.

Levels of carotene, vitamin A, and vitamin C measured in the serum of patients with cancer of the breast and uterine cervix were compared with levels in healthy controls and patients with benign diseases of the breast and cervix. Serum ascorbate levels were significantly lower in patients with benign diseases of the breast and cervix than in controls. In cancer patients, there was a significant trend of lower serum vitamin levels with increasing stage of the disease.

Poorly differentiated pediatric head and neck neuroblastoma: a diagnostic dilemma.

Primary pediatric head and neck neuroblastoma is rare, with metastatic disease being the more common mode of involvement in this anatomic region. Poorly differentiated neuroblastoma initially presenting in head and neck locations presents a diagnostic dilemma, especially when evidence of primary disease in typical abdominal, retroperitoneal and thoracic sites is lacking. This tumor cannot easily be distinguished from the other common pediatric small round cell malignancies that may originate in or metastasize to the head and neck. Recent years have seen great strides in the immunohistochemical as well as cytogenetic characterization of certain pediatric small round cell lesions. A never before reported solitary, poorly differentiated neuroblastoma of the right parotid gland in a 20-month-old female is presented in order to familiarize the otolaryngologist with the modern diagnostic armamentarium available for the accurate characterization, and thus appropriate workup and treatment of this disease.

A new scale for assessing behavioral agitation in dementia.

The primary purpose of the present study was to develop a reliable and valid rating instrument for assessing treatment efficacy for behavioral problems in the cognitively impaired elderly. The Behavioral and Emotional Activities Manifested in Dementia (BEAM-D) Scale was developed for the operational assessment of troublesome and disruptive behaviors in dementia. Each behavioral category of the BEAM-D was clinically considered to be a significant deviation from normative behavior for the geriatric dementia patient. The reliability and validity of the BEAM-D was assessed in a group of 45 patients diagnosed with primary degenerative dementia. The mean interrater reliability of BEAM-D items was 0.90. Concurrent validity was established by comparison with currently used rating scales, the Brief Psychiatric Rating Scale (BPRS) and the Sandoz Clinical Assessment-Geriatric (SCAG). Stepwise regression analysis revealed that the items of the BEAM-D had a strong relationship with conceptually similar behavioral dimensions on the BPRS and SCAG.

Augmentation of the effect of doxorubicin with low-dose tumor necrosis factor in experimental liver metastasis.

The antitumor activity of recombinant human tumor necrosis factor was studied in vivo as a single agent and in combination with a conventional chemotherapeutic agent. Dosages of tumor necrosis factor of 100 micrograms, 50 micrograms, and 25 micrograms were injected intraportally in Sprague-Dawley rats containing hepatic implants of Walker carcinosarcoma. An effect on the tumor was seen but was associated with a significant acute mortality. Lower dosages of tumor necrosis factor, 10 micrograms, 5 micrograms, and 1 microgram, administered with 10 mg/kg of doxorubicin (Adriamycin) significantly enhanced the antitumor effect of doxorubicin without an acute mortality. This suggests that lower dosages of tumor necrosis factor with conventional chemotherapy may augment the latter's effect without any added toxicity.

Immunological abnormalities in autoimmune chronic active hepatitis.

Autoimmune chronic active hepatitis is often associated with clinical and laboratory features that resemble those observed in systemic lupus erythematosus (SLE). We describe a 24-year-old woman with autoimmune chronic active hepatitis who was studied for serologic markers of autoimmunity and for immune clearance in terms of in vivo Fc receptor function. A markedly depressed immune clearance and splenic uptake of radiolabelled and IgG coated autologous erythrocytes was observed. The magnitude of this defect equaled or exceeded the most severe defects seen in a group of patients with SLE. This phenomenon was associated with markedly depressed serum C4 levels, a variably positive Sm antibody, and normal circulating immune complex concentrations. In addition, many clinical extrahepatic manifestations meeting criteria for classifying SLE were present. These findings further support the concept of autoimmune chronic active hepatitis and SLE being part of spectrum of overlapping autoimmune diseases.