Assessing immuno-expression of p53 protein and TP 53 gene amplification in histologically negative surgical margins of oral squamous cell carcinoma patients and normal oral mucosa.

OBJECTIVE: To assess and compare the immuno-expression of p53 and TP 53 gene amplification and correlate local recurrence and survival in histologically negative surgical margins of oral squamous cell carcinoma (OSCC) with normal oral mucosa. METHODS: Forty formalin-fixed paraffin-embedded tissue blocks of HNMs of OSCC and 40 normal oral mucosa samples were analyzed for p53 immunostaining and TP 53 gene amplification by PCR. RESULTS: Significantly, higher positivity was noted with p53 immuno-expression, TP53 gene amplification, and combined p53 and TP53 expression in the study group compared to the control group (C0.05). Most cases that were positive for p53 immuno-expression, TP 53 gene amplification, and combined p53 and TP53 expression showed local recurrence and poor survival. Kaplan-Meier survival analysis showed that subjects with TP53 and combined p53 and TP53 positivity had decreased survival rate than their negative counterparts. CONCLUSION: Detection of p53 in HNMs of OSCC can be used as a biomarker to identify patients at a higher risk of developing local recurrence and to predict survival. CLINICAL RELEVANCE: Combined p53 and TP 53 assessment may be more reliable for predicting LR to help clinicians and surgeons in treatment planning.

Caution with carotids: A necessary cognizance.

Most aberrations in the cervical course of the internal carotid artery are generally encountered as incidental findings due to their asymptomatic nature. However these morphological variations if not identified intra operatively or pre operatively may lead to vascular accidents. It is essential to be aware of the existence of tortuosity, kinking, and coiling of these great vessels while considering patients for surgeries of head and neck cancer, pharyngeal resections, traumatic injury repair, etc. Our paper presents a case series of four patients in whom carotid dolichoarteriopathies were observed during the course of neck dissection. It aims to highlight the precautions required to avoid morbidity in such patients.

Comparison of immunohistochemical expression of vascular endothelial growth factor and CD105 in oral squamous cell carcinoma: Its correlation with prognosis.

OBJECTIVES: Compare and correlate immunoexpression of vascular endothelial growth factor (VEGF) and CD105 in oral squamous cell carcinoma (OSCC), and correlate its expression with the prognosis of the patient. MATERIALS AND METHODS: A retrospective study was carried out on total of 49 cases of OSCC. Detailed demographic and clinical data were obtained, and tissue sections were stained with hematoxylin and eosin to grade the tumor. Later each case was subjected for immunohistochemical analysis of CD105 and VEGF. RESULTS: All the cases showed positivity for both CD105 and VEGF but high expression was noted with CD105 compared to VEGF. Average microvascular density for CD105 was higher (69.5) in moderately differentiated squamous cell carcinoma (MDSCC) when compared to well differentiated squamous cell carcinoma (WDSCC) (52.16). When expression of CD105 and VEGF was compared in WDSCC and MDSCC, it was statistically insignificant. However when expression of CD105 and VEGF was compared with survival of the patient, survival rate was <2 years in CD105 and was statistically significant, but VEGF did not show any significant difference with survival rate. CONCLUSION: CD105 immunoexpression in OSCC predicts a poor outcome than VEGF. So it can be postulated that endoglin may have a particular role in the development of cancer and might be relatively more specific than commonly used endothelial markers for squamous cell carcinoma of the oral cavity.

Estimation of the effects of normal tissue sparing using equivalent uniform dose-based optimization.

In this study, we intend to estimate the effects of normal tissue sparing between intensity modulated radiotherapy (IMRT) treatment plans generated with and without a dose volume (DV)-based physical cost function using equivalent uniform dose (EUD). Twenty prostate cancer patients were retrospectively selected for this study. For each patient, two IMRT plans were generated (i) EUD-based optimization with a DV-based physical cost function to control inhomogeneity (EUDWith DV) and (ii) EUD-based optimization without a DV-based physical cost function to allow inhomogeneity (EUDWithout DV). The generated plans were prescribed a dose of 72 Gy in 36 fractions to planning target volume (PTV). Mean dose, D30%, and D5% were evaluated for all organ at risk (OAR). Normal tissue complication probability was also calculated for all OARs using BioSuite software. The average volume of PTV for all patients was 103.02 ± 27 cm(3). The PTV mean dose for EUDWith DV plans was 73.67 ± 1.7 Gy, whereas for EUDWithout DV plans was 80.42 ± 2.7 Gy. It was found that PTV volume receiving dose more than 115% of prescription dose was negligible in EUDWith DV plans, whereas it was 28% in EUDWithout DV plans. In almost all dosimetric parameters evaluated, dose to OARs in EUDWith DV plans was higher than in EUDWithout DV plans. Allowing inhomogeneous dose (EUDWithout DV) inside the target would achieve better normal tissue sparing compared to homogenous dose distribution (EUDWith DV). Hence, this inhomogeneous dose could be intentionally dumped on the high-risk volume to achieve high local control. Therefore, it was concluded that EUD optimized plans offer added advantage of less OAR dose as well as selectively boosting dose to gross tumor volume.

Clinico-pathological factors affecting lymph node yield in Indian patients with locally advanced squamous cell carcinoma of mandibular Gingivo-Buccal sulcus.

AIM: Lymph node yield (LNY) is a valid marker of prognosis in oral cancer. Precise estimation of LNY in Indian patients with T3/T4 gingivobuccal sulcus squamous cell carcinoma (GBS-SCC) has not been well documented. Hence, the primary objective of the study was to determine the LNY in patients with T3/T4 SCC of mandibular GBS, and the secondary objective was to study the association of LNY with clinicopathological factors such as tumor thickness, histological differentiation, number of positive nodes, and extracapsular spread (ECS). MATERIALS AND METHODS: Study patients comprised biopsy proven T3/T4 SCC of mandibular GBS that underwent unilateral surgery (composite or bite composite resection with level I to level V-neck dissection and pectoralis major flap reconstruction) at our center between January 2012 and October 2014. Grossing of surgical specimens was done as per the guidelines established by the Royal College of Pathologists (December 2009). The data were analyzed using SPSS software (22nd version) and Chi-square test. RESULTS: The surgical specimens of 106 patients yielded 2329 lymph nodes with the mean LNY of 21.97 ± 5.57. Higher mean LNY of over 21 was significantly associated with ECS, number of positive nodes, delay in surgery over 15 days, skin involvement by the tumor, and presence of oral potentially malignant disorders. CONCLUSION: With the single surgeon, pathologist and same surgical procedure, the mean LNY in Indian patients with T3/T4 SCC of mandibular GBS is 21.97 ± 5.57. Although clinicopathological factors affect the estimation of LNY, further studies are needed to validate the findings of this study.

A tecnique to maximise length of pectoralis major muscle myocutaneous flap pedicle in orofacial reconstruction.

Introduction Pectoralis muscle flap is one of the most common regional flap used in orofacial reconstruction. Mobilization of pedicle and reach of the skin paddle is a technical consideration. Method Based on the anatomical difference in blood supply to clavicular and sternal head, we elaborate a tecnique to maximise the length of pedicle in maxillofacial reconstruction. Conclusion The proposed technique is reproducible and reliable and possibly increase the indications of usage of the flap.

Mandibular peripheral primitive neuroectodermal tumor: a rare case report with review of literature.

Primitive neuroectodermal tumor (PNET) is a high grade malignant neoplasm of small round cell tumor family, commonly affecting children and young adults. Peripheral primitive neuroectodermal tumor (pPNET) is a predominately neural, nonepithelial malignancy seen outside the nervous system that can arise in any place throughout the body including the diverse tissues of the head and neck. The diagnosis of PNET is confounded by its clinical and histopathological similarity to Ewing's sarcoma of the bone and has seldom been reported in the literature. The paucity of literature pertaining to the successful diagnosis and management of this lesion mandates its documentation and discussion. This article describes a case of an 11-year-old boy with an aggressive pPNET of the mandible. The clinical and radiographic presentations of this rare entity along with a detailed review on the current management modalities have been discussed.

Modification of MacFee Incision: A Balance Between Cosmesis and Accessibility.

Incision design is a critical element of operative planning. Incisions are chosen with the aim of optimizing exposure of relevant neck levels and minimizing morbidity. Various incisions have been proposed over the years but none fulfilled the criterias of ideal incisions. We propose a modification of MacFee s incision which is the balance between cosmesis and exposure.

Regrafting of nipple areolar complex during pectoralis major myocutaneous flap reconstruction.

Pectoralis major myocutaneous flap is one of the most popular reconstructive methods employed in head and neck reconstruction.cosmesis and symmetry are important components of any surgery. Here we report a simple and effective method to restore cosmesis and symmetry.

Recombinant UL30 antigen-based single serum dilution ELISA for detection of duck viral enteritis.

A recombinant UL30 antigen-based single serum dilution enzyme linked immunosorbent assay (ELISA) was developed to measure specific antibody in the sera of ducks against duck enteritis virus (DEV). The partial UL30 gene of DEV was cloned, expressed, purified and tested for its diagnostic use by designing a single serum dilution enzyme linked immuno-sorbent assay (ELISA). A total of 226 duck sera samples were tested using the assay. A linear relationship was found between the predicted antibody titres at a single working dilution of 1:100 and the corresponding serum titres observed as determined by the standard serial dilution method. Regression analysis was used to determine a standard curve from which an equation was derived which demonstrated this correlation. The equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay proved to be specific, sensitive and accurate as compared to the virus neutralization test with a specificity, sensitivity and accuracy being 96%, 95% and 95% respectively.

Oral microbial carriage in oral squamous cell carcinoma patients at the time of diagnosis and during radiotherapy - a comparative study.

OBJECTIVES: Tobacco chewing habit, presence of squamous cell carcinoma in oral cavity and radiotherapy causes alterations in healthy oral microflora. Abnormal flora developed due to radiotherapy in oral squamous cell carcinoma (OSCC) patients can exacerbate mucositis and can cause systemic infections. The role of oral microorganisms in carcinogenesis is gaining interest recently. Abnormal flora in development of second tumor in the field of first tumor is to be established. The study fundamentally tries to evaluate the shift that occurs during the radiotherapy in OSCC patients. METHODS: Microbial analysis of saliva samples from OSCC patients undergoing radiotherapy, tobacco chewers and controls was undertaken. The microorganisms were grouped into categories as total aerobes, total anaerobes, candida, coliforms and gram negative anaerobic bacteria. RESULTS: The frequency of isolation of total aerobes, total anaerobes, coliforms and gram negative anaerobic bacteria was significantly high in OSCC patients compared to healthy controls whereas candida was isolated most frequently during radiation period. The tobacco chewers showed significant increase in colony forming units of total aerobes and coliforms. All the microbial groups were high in OSCC and radiotherapy patients. While OSCC patients showed significant increase in total anaerobes and gram negative anaerobes, candida was increased in radiotherapy patients only. CONCLUSION: Habits promote coliforms. Tumor supports efficiently anaerobes and candida. The latter is supported more by radiation. The study stresses the importance on administration of appropriate antimicrobial therapy right at the time of diagnosis of the lesion.

Soft tissue reconstruction after hemimandibulectomy in oral malignancies: a retrospective study.

AIMS: The present study was under taken to evaluate the efficacy of various local and regional soft tissue flaps used for reconstruction after excision of various malignant lesions of the mouth and also to evaluate complications with length of hospital stay after the reconstruction. MATERIALS AND METHODS: The study was a record based retrospective analysis of 127 patients who were histologically proven squamous cell carcinoma of the oral cavity for which excision of the lesion along with segmental mandibulectomy and primary reconstruction with local or regional flaps was the treatment modality. RESULTS: The male:female mean age is 48.27:48.79. The Z-proportionality test for intra oral reconstruction showed 5% level of significance (P < 0.05) between pectoralis major myocutaneous flap (PMMC) and other flaps. Difference between deltopectoral (DP) and PMMC, PMMC and primary closure at 1% level of significance, i.e. P < 0.01 was found for extra oral defects. The mean stay was found to be 31.31 days. Recurrence rate of 11% was reported. CONCLUSION: A total of 127 patients formed the study group. In the absence of bone reconstruction PMMC still continues to be the "work horse" of reconstruction following wide excision and hemimandibulectomy.

Evolutionary dynamics of the American African genotype of dengue type 1 virus in India (1962-2005).

Dengue is a major health problem in India with all four serotypes represented. Recently there has been an increase in the occurrence of dengue-1 outbreaks. It is possible that there have been changes in the genetics of dengue virus-1 (DENV-1), either by fresh introductions or by evolution in situ. The studies on DENV-1 evolution so far have no Indian sequences included. To gain insight into the dynamics of DENV-1 in India, the envelope (E) gene of thirteen virus isolates representative of the period 1962-2005 were sequenced and analyzed together with the available sequences of 40 globally representative isolates. All the Indian DENV-1 isolates were found to belong to the American African (AMAF) genotype. With the addition of 13 Indian isolates, the AMAF genotype can now be called Cosmopolitan. The Indian isolates were distributed into four lineages, India I, II, III and the Africa lineage, now called Afro-India. Of these, India III was the oldest and extinct lineage; the Afro-India was a transient lineage while India I, imported from Singapore and India II, evolving in situ, were the circulating lineages. Despite the extinction and introduction of lineages, no specific codon site was observed to be under selection pressure. The rate of nucleotide substitution estimated for DENV-1 was 6.5 × 10(-4) substitutions/site/year, and the time to the most recent common ancestor (tMRCA) was estimated to be 78-180 years (1825-1925), similar to previous estimates. The tMRCA for the AMAF/Cosmopolitan genotype was 56-98 years (1907-1949), a period that covers World War I and II. The two imports from Africa (1953-1968) and Singapore (1964-1975) and an export to the Americas (1955-1965) prove that there have been changes in the lineage of the DENV-1 viruses circulating in India which has contributed to the global dynamics of DENV-1 evolution and perhaps to the changing epidemiology of dengue in India.

Prevalence and distribution of high risk human papillomavirus (HPV) Types 16 and 18 in Carcinoma of cervix, saliva of patients with oral squamous cell carcinoma and in the general population in Karnataka, India.

BACKGROUND: In view of conducting HPV vaccination in India it is most important to understand the prevalence of HPV genotypes in this population, not only in squamous cell carcinoma of cervix and oral cavity but also in the general population. In this study we explored the prevalence and distribution of high-risk HPV types 16 and 18 in carcinoma of cervix, saliva of patients with oral squamous cell carcinoma and in general population in Karnataka. METHODS: Cervical cancer specimens after punch biopsy (n=60) were obtained from women attending Karnataka Institute of Medical Sciences and Karnataka Cancer Therapy and Research Institute, Hubli (KCTRI). Saliva rinse of (n=34) OSCC patients from KCTRI and (n=396) normal individuals from different regions of North Karnataka, were collected and PCR based high-risk HPV genotyping was carried out. RESULTS: Using consensus PCR primers it was observed that 96.7% patients were infected with HPV irrespective of specific type in cervical cancer. Among them, HPV 16 was observed in 89.7%, HPV 18 in 86.2% and both HPV 16 and 18 in 79.3% patients. In OSCC, 70.6% were positive for HPV, among which HPV 16 prevalence was observed in 45.8%, HPV 18 in 54.2%, and HPV 16 and 18 multiple infection in 4.18%. In general population, HPV prevalence was observed in 84.4%. Among them, HPV 16 was observed in 2.75% and HPV 18 in 22.0% patients. In general population, multiple infection with HPV 16 and 18 was not observed but 75.3% were found to be infected by HPV genotypes other than HPV 16 and 18. CONCLUSIONS: Our study reveals that multiple infection of HPV 16 and 18 is quite high in cervical cancer and in case of OSCC, it was in conformity with the other studies. In general population HPV 18 prevalence was observed to be high. With this, we can conclude that both HPV 16 and 18 vaccinations will reduce the burden of cervical cancer and OSCC in Karnataka.

Prevalence and risk of leukemia in the multi-ethnic population of North Karnataka.

BACKGROUND: The aim of this study was to analyze the trends in the incidence rates of four major types of leukemia in the population of North Karnataka, which accounts for the 2.5% of the whole population of India. Due to the lack of any nationwide leukemia screening program, the majority of the people are not aware of the disease. Epidemiological study can play a vital role in understanding the occurrence and outcome of the disease. PATIENTS AND METHODS: Focusing on variables like age, sex, race, blood group and lifestyle habits, detailed reports of 417 males and 230 females (M:F ratio 1.8:1) were collected from different hospitals of North Karnataka and analyzed for their risk of leukemia. RESULTS: Compared to female patients, Hindu males were found to have greater risk of occurrence of leukemia (p=0.0333). The males of scheduled caste (SC) and Lingayat communities showed a high risk than other communities (p=0.000). The occurrence of AML showed a significant relationship with ABO blood groups (p=0.0090). The frequency of leukemia is quite high in Belgaum district when compared to others districts of North Karnataka and totally absent in Bidar district. The reasons need precise molecular and genetical studies of the population. CONCLUSIONS: The localized communities of Lingayat and SCs needs to be further studied to get a better understanding of the higher risk of occurrence of leukemia in males. Moreover, since the spectrum of cancer epidemiology seen in India is different from that in developed countries more emphasis should be placed on better development of regional and national registries.

Single prediction equation for bioelectrical impedance analysis in adults aged 22-59 years.

The purpose of this study was to validate a single bioelectrical impedance analysis (BIA) equation in healthy Indian subjects aged 22-59 years with a body mass index (BMI) between 16.8 and 47.3 kg m⁻². Healthy subjects (34 men and 30 women) were measured by two methods: bone mineral content (BMC) was measured by a commercial body composition analyser and bioelectrical impedance at various frequencies was measured by a newly developed bioelectrical impedance measurement system. As correlations were high and prediction error was low, a single equation was developed using all subjects as follows: BMC = -3.5268 + (0.0279  x  h) + (0.0145  x  w) + (184  x  (h²/Z(body50)))-(1.08  x  (w x  h²/Z(body6.25)))-(0.0032  x  (age))-(0.103  x  (sex); men = 1, women = 0). BMC measured from commercial instrument InBody720 was 2.552 ±â€Š0.457 kg. BMC predicted by equation was 2.554 ±â€Š0.447 kg (R = 0.976, adjusted R²â€Š= 0.948, standard error of estimate = 0.104 kg, total error = 0.09987 kg). The results of this study show that the newly developed multi-frequency bioelectrical impedance measurement system with the single prediction BIA equation can be used in screening the subjects suspected with osteoporosis and for follow-up study of the patient under the therapy for osteoporosis. For validation of commercial instrument InBody720, BMC of 22 healthy subjects was measured by InBody720 and dual-energy X-ray absorptiometry. High correlation (R = 0.9531) and low error (total error = 0.0913 kg) was found between these two methods.

Histological significance of p53 gene expression in squamous cell carcinoma of the buccal mucosa.

INTRODUCTION: Oral Squamous Cell Carcinoma (SCC) results from genetic mutations which activate the oncogenes and inactivate the tumor suppressor gene namely TP53. Despite the use of multimodality treatments the prognosis of oral SCC has not changed significantly. PURPOSE: To evaluate 1) if there is any correlation between the two prognostic indicators i.e. p53 over expression and histological grade of the tumor 2) if any of the parameters of histological grading correlate significantly with p53 over expression. This information would help in understanding the exact role of TP53 gene mutation in cellular progression of oral SCC. METHOD: Study was conducted on 90 resected specimens of Stage IV SCC of buccal mucosa. Slides from these specimens were evaluated for histological grading by Anneroth's method and p53 over expression by Immunohistochemistry. RESULTS: Statistically significant co-relation was seen between the total histological grade and p53 over expression. Also 4 individual histological parameters which indicated high cellular turnover were also significantly associated with p53 over expression. CONCLUSION: TP53 mutation histologically signifies an early event in cellular progression of oral SCC.

Characterization of porcine stable kidney cell line adapted to hyperthermic temperature.

The optimum temperature for the growth of porcine stable (PS) kidney cell line is 37 degrees C. We have adapted the cell line to grow at 40 degrees C. The original cell line grown at 37 degrees C has been denoted as PS-37, and the adapted new strain has been denoted as PS-40. Both the cell lines were screened for mycoplasma by Hoechst staining and tritiated uridine-uracil uptake and were found to be negative. Comparative characterization of PS-40 and its progenitor PS-37 cell line was done by using various parameters. The antigenic studies indicated that the new cell strain was not cross-contaminated with any other cell lines. It was observed that PS-40 cells were more fibroblastic with clean cytoplasm and appeared healthy. The growth of PS-40 cells was faster than the original cell line. The karyological study showed heteroploid chromosome number in PS-40 cells. The modal chromosome number of PS-40 cells was 58, whereas that of PS-37 cell line was 38. The lactic dehydrogenase isoenzyme pattern showed a cathodal shift of bands. The PS-40 cell strain could be cryopreserved and revived. The viability of PS-37 as well as PS-40 cell lines is in the range of 90-95%, and the growth characteristics of thawed cells showed six- to eightfold multiplications within 5 d. The virus susceptibility study revealed that the cytopathic effect was more profound and observed 1 d earlier in PS-40 cell line. Increased yields of Japanese encephalitis, Sindbis, and Semliki forest viruses were obtained by 1.8, 1.75, and 1.5 log plaque-forming units/ml, respectively. The yield of West Nile virus was, however, comparable to that in PS-37 cell line. Both the cell lines were refractory to Dengue viruses.

Synthesis and antibacterial activity of some Schiff bases and 4-thiazolidinones.

Some new schiff bases (1a-d) 4-thiazolidinones (2a-d) have been synthesised and tested for their antibacterial activity. The structures of these compounds have been established on the basis of elemental analysis and spectral data (IR and H1 NMR).