The prognostic role of immunohistochemical markers in surgical margins of Indian patients with oral squamous cell carcinoma- a retrospective study protocol

<p><strong>Background:</strong> Positive surgical margins in oral squamous cell carcinoma (OSCC) is an important predictor of recurrence. Unfortunately, recurrence is found in cases with negative surgical margins as well and hence molecular markers are being investigated to identify the tumor cells in negative surgical margins to improve the survival of patients. The study aimed to ascertain the prognostic role of p53, eIF4E and E-cadherin in negative surgical margins and association of their expression with clinical parameters, recurrence, and survival.</p><p><strong>Methods:</strong> India based retrospective cohort study of patients who were surgically treated for OSCC between 2011 and 2014. Thirty patients with negative surgical margins were assessed for this study. Sixty-four negative surgical margins were evaluated using immunohistochemistry with monoclonal mouse p53, monoclonal rabbit eIF4E and monoclonal mouse E-cadherin antibodies.</p><p><strong>Results:</strong> Fisher’s exact test, Fishers Freeman Halton exact test and contingency tables were used to assess the association between recurrence rate and clinicopathologic parameters also any association with the expression of p53, eIF4E and E-cadherin in the surgical margins and each patients’ clinical characteristics. ANOVA (analysis of variance) was used to assess the role of clinicopathologic parameters in survival of the patients. The sensitivity and specificity of each antibody along with positive and negative predictive values were calculated. Kaplan Meier was used for analysis of recurrence and five-year survival. P&lt;0.05 was used to denote a statistically significant difference.</p><p><strong>Conclusion:</strong> This study could help in identification of patients with minimal residual cancer in the surgical margins and might help in better therapeutic management to improve survival of OSCC patients.</p>

Epidemiology of infections caused by Chlamydophila pneumoniae in patients with chronic cough

Background: Chlamydophila pneumoniae is an important etiological agent in respiratory system infections. The aim of study was to analyze the rate of Chlamydophila pneumoniae infection in adults and children and also to determine a correlation between the presence of this pathogen and symptoms of chronic cough. Material/Methods: The material for the study included swabs from the posterior pharyngeal wall taken on an empty stomach without cleaning the mouth. The diagnostic method was indirect immunofluorescence test (IIFT), which uses two types of antibodies: monoclonal mouse antibodies, which link specifically with the antigen that is present in the tested material and goat anti-mouse antibodies linked to fluorescein isothiocyanate, providing the colour reaction with C. pneumoniae antigen. Results: In our research, 593 patients, including 319 women, 175 men, aged from 18 to 87 years and a group of 99 children aged from 2 to 17 years with symptoms of chronic cough n=432 and other respiratory manifestations n=161 were studied. In the group of studied women with cough, 28.2% (64/227) of results were positive. In the group of men with cough, 22.3% (27/121) of results were positive. In the group of children with a cough, 28.6% (24/84) of the results were positive. Conclusions: In the examined group of children and adults with a chronic cough, the C. pneumoniae antigen was detected. The frequency of detection of C. pneumoniae antigen differed depending on the age group of both children and adults with symptoms of chronic cough.

Assessment of Expression of Ki-67 in Benign and Malignant Prostatic Lesions among Sudanese Patients

BACKGROUND: Prostatic cancer is one of the most common cancers of males in a Sudanese population. The early detection is very important, as it is only curable at an early stage. AIM: The objective of this study was to investigate the expression pattern of Ki-67 in benign and malignant prostatic lesions to improve the diagnosis that may help in better management and prevention of disease. MATERIAL AND METHODS: Fifty-eight formalin fixed paraffin blocks from diagnosed cases of prostatic tumours with different grade, and stages were included in this study. Ki-67 expression was examined immunohistochemically by using monoclonal mouse anti-human Ki67 IS626. The results were correlated with Gleason score and tumour differentiation and stage. RESULTS: The frequency of histological types was as follow: 11 cases of benign prostate,atic hyperplasia (19%) and 47 cases of prostatic cancer (81%). Our results stated that prostatic adenocarcinoma among Sudanese patients was of low grade which means tumours are less aggressive. Furthermore, the findings demonstrate that Ki-67 expression in prostatic carcinoma smears was correlated significantly with the degree of Gleason score (P < 0.05). CONCLUSIONS: We found that the prostatic adenocarcinoma among Sudanese patients was less aggressive. Furthermore, Ki-67 expression was proportional to the grade of a tumour and it was a useful prognostic and diagnostic biomarker.

Thalamic Reticular Nucleus in Caiman crocodilus: Immunohistochemical Staining

The thalamic reticular nucleus in reptiles, Caiman crocodilus, shares a number of morphological similarities with its counterpart in mammals. In view of the immunohistochemical properties of this nucleus in mammals and the more recently identified complexity of this neuronal aggregate in Caiman, this nucleus was investigated using a number of antibodies. These results were compared with findings described for other amniotes. The following antibodies gave consistent and reproducible results: polyclonal sheep anti-parvalbumin (PV), monoclonal mouse anti-PV, and polyclonal sheep anti-glutamic acid decarboxylase (GAD). In the transverse plane, this nucleus is divided into two. In each part, a compact group of cells sits on top of the fibers of the forebrain bundle with scattered cells among these fibers. In the lateral forebrain bundle, this neuronal aggregate is represented by the dorsal peduncular nucleus and the perireticular nucleus while, in the medial forebrain bundle, these parts are the interstitial nucleus and the scattered cells in this fiber tract. The results of this study are the following. First, the thalamic reticular nucleus of Caiman contains GAD(+) and PV(+) neurons, which is similar to what has been described in other amniotes. Second, the morphology and distribution of many GAD(+) and PV(+) neurons in the dorsal peduncular and perireticular nuclei are similar and suggest that these neurons colocalize these markers. Third, neurons in the interstitial nucleus and in the medial forebrain bundle are GAD(+) and PV(+). At the caudal pole of the thalamic reticular nucleus, PV immunoreactive cells predominated and avoided the central portion of this nucleus where GAD(+) cells were preferentially located. However, GAD(+) cells were sparse when compared with PV(+) cells. This immunohistochemically different area in the caudal pole is considered to be an area separate from the thalamic reticular nucleus.

Evaluation of tumor marker cancer antigen 72-4 (CA 72-4) in the monitoring of metastatic or recurrent tumors of the gastrointestinal tract, lung, breast, and ovaries.

263 Background: In management of metastatic and recurrent cancers, measuring response is a constant challenge. CA72-4 is a tumor marker (TM) that has been found elevated in a variety of human adenocarcinomas, with reported sensitivities of up 50% and overall specificity of over 95%. Using the DRG TM-CA72-4 assay, we quantified the abnormality rate of TM CA72-4 compared with current FDA-approved TM in various cancers. Methods: We conducted a prospective, single center study by enrolling 96 patients between March 2013 and August 2016 with various de novo or previously diagnosed locally advanced, unresectable and/or metastatic cancers known to express CA72-4. Quantification of CA72-4 was performed according to manufacturer’s instructions using the DRG TM-CA72-4 ELISA kit, which was developed by DRG International (Germany) utilizing the CC-49 monoclonal mouse antibody directed against an epitope on the CA72-4 antigen. Positivity was calculated as greater than A) 0.8 U/mL or B) 4.0 U/mL based on systematic review of prior studies. Results: The positivity rates based on different cut-off points (0.8 U/mL vs 4 U/mL) and their corresponding FDA approved tumor markers are shown in the Table. Conclusions: Positivity rates of CA72-4 varied based on different assay cut-off levels with the highest positivity noted in the pancreatic, ovarian and colorectal carcinomas indicating a potential role for disease monitoring. [Table: see text]

A portable and quantitative enzyme immunoassay of neuron-specific enolase with a glucometer readout

A portable and quantitative enzyme immunoassay with a glucometer readout was developed for the sensitive monitoring of neuron-specific enolase (NSE, as a model analyte) in a high-binding polystyrene 96-well microtiter plate (MTP), conjugated with monoclonal mouse anti-human NSE antibody (mAb1).

Immunohistochemical distribution of prolactin containing cells in the pituitary of the chickens

The objective of the current study was to determine the immunohistochemical distribution and density of prolactin containing cells in the pituitary of the female and male chickens during postnatal developing period after hatching for five months. The modified labelled avidin-biotin method with monoclonal mouse antihuman prolactin as a primary antibody was used to detect prolactin positive cells. At the end of each month, prolactin containing cells were identified in the pars distalis of the pituitary glands of chickens. The number of the prolactin positive cells were found to be increased depending on the monthly development period, while the reaction density was found to be decreased in all groups. Additionally, males had less prolactin positive cells than that of females.

Determination of metallothionein in biological fluids using enzyme-linked immunoassay with commercial antibody.

Metallothionein (MT) is a low molecular weight cysteine-rich protein with a number of roles in the pro/antioxidant balance and homeostasis of essential metals, such as zinc and copper, and in the detoxification of heavy metals, such as cadmium and mercury. Until now, detection of metallothionein in biological fluids remained difficult because of a lack of a broadly reactive commercial test. Meaningful comparison of the values of metallothionein concentrations reported by different authors using their specific isolation procedures and different conditions of enzyme-linked immunoassay is difficult due to the absence of a reference material for metallothionein. Therefore in the present study, we describe a quantitative assay for metallothionein in biological fluids such as plasma and urine performed by a direct enzyme-linked immunoassay using a commercially available monoclonal mouse anti-metallothionein clone E9 antibody and commercial standards of metallothionein from rabbit liver and a custom preparation of metallothionein from human liver. The sensitivity of the assay for the standard containing two isoforms MT-I and MT-II from human liver was 140 pg/well. The reactivity of the commercial standards and standards containing two isoforms MT-I and MT-II isolated from human liver in our laboratory with a commercial monoclonal mouse anti-metallothionein clone E9 antibody were similar. This suggests that the described ELISA test can be useful for determination of metallothionein concentration in biological fluids. The concentrations of metallothionein in human plasma, erythrocyte lysate and in urine of smoking and non-smoking healthy volunteers are reported. Tobacco smoking increases the extracellular metallothionein concentration (plasma and urine) but does not affect the intracellular concentration (erythrocyte lysate).