Development of a sensitive cost-effective capture ELISA for detection of murine monoclonal antibodies: correlation with SPR biosensor technology.

The development of antibodies for diagnostic and therapeutic applications in inflammatory diseases is a major focus for biotechnology and pharmaceutical companies. Production of monoclonal antibodies requires the development of fast, high-throughput methodologies for screening and selecting appropriate candidate antibodies for development. Capture (sandwich) enzyme linked immunosorbent assay (ELISA) provides a quick and reliable method that could be used for hybridoma screening of potential candidates accompanied with surface plasmon resonance (SPR) biosensor technology for identifying high affinity biomolecular interactions. A sensitive, cost-effective, robust and accurate capture ELISA for detection of murine monoclonal antibodies in culture supernatants was developed. This assay was optimized for high sensitivity and specificity with a capture anti-mouse polyclonal antibody. Using serial dilutions of a defined murine IgG antibody, a linear dose-response was observed between 2 and 1200 ng/ml antibody with a coefficient of determination r2 of 0.98. The detection limit of the assay was established as 2ng/ml (12.5pM). A similar concentration-dependent doseresponse was also observed using serial dilutions of antibody-containing supernatants from anti-alpha glycophorinproducing hybridomas (detection limit 1:2000). Specific capture of antibodies from supernatants in a similar setting was also confirmed using SPR biosensor technology and correlated well with the immunoassay results. The latter technology can be performed in order to provide quick screening results and kinetic analysis of antibody binding interactions aiming at identifying candidates with high affinity and specificity.

An unexpected pulmonary arterial aneurysm in a COPD patient.

We present a case of an idiopathic pulmonary artery aneurysm in an asymptomatic patient who was treated for an irrelevant medical condition. Pulmonary artery aneurysms (PAA) are quite rare and can either be congenital or acquired. Congenital aneurysms are usually associated with cardiac malformations leading to pulmonary hypertension. Acquired aneurysms can be idiopathic or associated with infections (tuberculosis, syphilis), trauma, pulmonary valvular stenosis, or collagen diseases. Pulmonary artery aneurysms are not common and an idiopathic pulmonary artery aneurysm is a rare finding that could be diagnosed incidentally.

Miliary pattern due to occupational lung disease in a patient with laryngeal cancer.

Inhalation of metal dusts and fumes can induce a wide range of respiratory disorders, including granulomatosis, chemical pneumonitis and pulmonary interstitial disease. Laryngeal cancer is the most common cancer of the upper aerodigestive tract. We present a patient with occupational lung disease whose chest CT showed miliary nodular pattern, with concurrent laryngeal cancer who had been engaged in type printing for 22 years. Histology of the laryngeal lession showed squamous cell laryngeal cancer. Histology of the nodules showed a foreign body granulomatous response with several foreign body cells, most probably due to exposure to numerous inorganic (lanthanides, elements such us La, Ce, Nd, Sm, EU, Tb, Lu) and organic particles (such us acrylates, epoxy- and urethane-acrylates).

Hormonal responses to marathon running in non-elite athletes.

BACKGROUND: Exercise is known to be a powerful stimulus for the endocrine system. The hormonal response to exercise is dependent on several factors including the intensity, duration, mode of exercise (endurance versus resistance), and training status of the subject. The aim of the present study was to determine the steroid hormonal response (immediately after a race and 1 week later) to endurance exercise under the real conditions of the classic Athens marathon in a group of well-trained, middle-aged, non-elite athletes. METHODS: Blood samples were drawn 1 week before the race, directly after completion of the race, and 1 week later. RESULTS: Serum cortisol and prolactin showed distinct rises 1 h after the race and returned to baseline 1 week later. Androstenedione and dehydroepiandrosterone sulphate did not show any changes. Total testosterone as well as free testosterone dropped significantly 1 h after the race but returned to baseline 1 week later. CONCLUSION: In this particular group of non-elite, middle-aged marathon runners, the race resulted in an acute increase in serum cortisol and prolactin levels and in a concomitant decline in testosterone level. The aforementioned changes returned to baseline 1 week later.

Pulmonary complications in diabetes mellitus.

Clear decrements in lung function have been reported in patients with diabetes over the past two decades, and many reports have suggested plausible pathophysiological mechanisms. However, there are no reports of functional limitations of activities of daily living ascribable to pulmonary disease in patients with diabetes. This review attempts to summarize the available information from the present literature, to describe the nature of the lung dysfunction in diabetes and the emerging clinical implications of such dysfunction.

Periodic limb movement during sleep and chronic obstructive pulmonary disease.

The aim of this work was to study whether chronic obstructive pulmonary disease (COPD) subjects exhibited periodic limb movement (PLMs) during sleep. A retrospective case control study was conducted in the referral sleep disorders laboratory in the University of Patras in southwest Greece. A sample of 23 COPD subjects was compared with 14 severe obstructive sleep apnea (OSA) subjects and 18 periodic limb movement disorder (PLMD) subjects. The PLM Index (PLMI) and PLMs Arousal Index (PLMAI) in COPD subjects differ (p<0.05) from severe OSA patients. The PLMAI differ (p < 0.05) between COPD and PLMD subjects. Spearman's correlation showed a positive statistical significant correlation between PLMI and PLMAI in the entire population and in COPD subjects. There was no statistical significant correlation between sleep-related symptoms and the occurrence of PLMs disorder in COPD patients. In our study, PLMs with associated arousals are often seen in COPD subjects. Further prospective studies will be necessary to clarify the mechanisms whereby the reduction in PLMs in COPD patients improved their sleep and quality of life.

Indications, results and complications of flexible fiberoptic bronchoscopy: a 5-year experience in a referral population in Greece.

The aim of this study was to retrospectively review the indications, results and complications of flexible fiberoptic bronchoscopy (FFB) in an University teaching Hospital. Also, we present the radiological findings for the major causes according to computed tomography of the chest performed within 48 h of fiberoptic bronchoscopy. A total of 4,098 FFBs were performed from January 1, 2003 to December 30, 2007. For diagnostic purposes, 3769 FFBs performed (92%) and for therapeutic purposes 329 FFBs (8%) performed. Haemoptysis was the most common indication for FFB (21%), followed by fever/suspected infection (19%) and chronic cough (18%). The most common results of the diagnostic workup was nonspecific inflammation of the tracheobronchial tree (31% for haemoptysis, 38.7% for fever and 48.5% for chronic cough), with malignancy ranking second (17%, 26.1% and 26% respectively). The cytological results showed adenocarcinoma to be the most common lung cancer in both sexes (37.3% for men and 39.7% for women). The mortality rate was 0.04% and the frequency for major and minor complications was 0.56% and 0.33%, respectively. In conclusion, flexible fiberoptic bronchoscopy is a safe procedure and can play a major role in both diagnosis and treatment, as long as the requisites of preparation and supervision are followed.

Specific sensitization in wheat flour and contributing factors in traditional bakers.

Skin prick tests (SPTs) play an important role in the diagnosis of bakers asthma and epidemiological field studies on frequencies of sensitization to wheat or rye flour. In many epidemiological studies the investigators have tried to reveal the determinants of specific sensitization, and atopy appears to be a very strong determinant for sensitization to flour allergens. Age and gender have not reported to be determinants of sensitization. Only one study found that sensitization in wheat flour with skin prick test was significantly associated with cigarette smoking. Few data are available on sensitization to wheat flour in populations without occupational exposure to bakery allergens. The aim of this study was to investigate the sensitization of wheat flour and other baking allergens (oat, barley, and rye flour) in traditional bakers and in cleaners in a big hospital in the same area. In addition the correlation of atopy with specific sensitization and the correlation of specific sensitization with age, sex, working hours, working years, smoking status and pack/years with use of skin prick tests.

Sleep apnea syndrome in a referral population in Greece: influence of social factors.

OBJECTIVE: The aim of this work was to study whether social factors are risk factors for obstructive sleep apnea (OSA). A second objective was to investigate gender differences in relation to referral to a sleep laboratory for sleep-related breathing symptoms. STUDY DESIGN: A retrospective cross-sectional study was conducted in the referral sleep disorders laboratory in the tertiary University Hospital in Patras in southwest Greece. A sample of 362 subjects originated from this geographic region was screened for social characteristics, i.e., marital status, occupation, and education. RESULTS: The apnea hypopnea index (AHI) was approximately three times as high in men as in women (p < 0.05). Snoring was reported to be a symptom by 76.6% of males and 75% of females. Excessive daytime sleepiness (EDS) was reported by 25.5% of males and 15% of females. Arousals during sleep were reported by 5.7% of men and 10% of women. The ratio of subjects with concomitant disorders or symptoms did not differ between sexes (p > 0.05). The influence of age, body mass index (BMI), gender, smoking, and social characteristics on AHI was examined by multinomial logistic regression. The following factors remained independent risk factors for the presence of moderate to severe OSA (i.e., AHI > 15/h compared with AHI < 5): (1) Gender: the odds ratio (OR) of males to females was 6.23 (CI = 1.89-20.5). (2) Obesity: the OR of subjects with BMI >30.5 kg/m(2) in comparison to those with lower BMI was 3.83 (CI = 1.86-7.86). (3) Marital status: The OR of married subjects to singles was 2.30 (CI = 1.01-5.32). (4) Occupational status: The OR of subjects outside the work force was 3.85 (CI = 1.16-12.74) and that of the self-employed was 1.70 (CI = 0.70-4.10) compared to a reference group of clerks/employees. CONCLUSION: In our study factors associated with the presence of sleep apnea include gender (men), obesity, marriage, and self-employment or being outside the work force.

Urinothorax: an unexpected cause of pleural effusion in a patient with non-Hodgkin lymphoma.

Urinothorax is a rather rare cause of pleural effusion and its potential mechanism is urinary tract obstruction or trauma that results in urine leakage and accumulation inside the pleural space. Patients with non-Hodgkin lymphoma could present with pleural effusion due to mediastinal lymphadenopathy or extrathoracic manifestation such as urinary tract obstruction, the latter described in our case report. Physicians must be aware even of the more occult mechanisms of pleural fluid accumulations which could point to extrathoracic manifestations of involvement.

Does CPAP therapy improve erectile dysfunction in patients with obstructive sleep apnea syndrome?

OBJECTIVE: The aim of the study was to examine the quality of the characteristics of Erectile Dysfunction (ED) in men with Obstructive Sleep Apnea Syndrome (OSAS) and to investigate whether there is an improvement with the use of continuous positive airway pressure (CPAP) therapy. MATERIALS AND METHODS: Fifteen men with OSAS and sexual dysfunction have been investigated in this follow-up study. The treatment period was for 12 weeks and the therapeutic CPAP levels were determined during the full night of the therapeutic titration. RESULTS: In our 15 treated patients, the International Index Erectile Function (IIEF) total and all domain scores had increased after the CPAP treatment compared to the baseline, except for that of sexual desire domain. CONCLUSIONS: CPAP therapy can improve the sexual function in ED patients with OSAS.

Carcinoid tumour behind bronchiectasis.

This report describes a female patient with bronchiectasis, presented to our department with recurrent hemoptysis. Bronchoscopy revealed nothing else but blood arising from the upper lobe bronchus. High resolution computing tomography of the lung (HRCT) revealed bronchiectasis of the upper lobe. A right upper lobectomy was performed. Behind bronchiectasis multiple nodular lesions, 5-10 mm were observed. Histological and immunohistochemical examination revealed findings consistent with peripheral typical bronchial carcinoids.

Accuracy of pulmonary function tests in predicted exercise capacity in COPD patients.

PURPOSE: The purpose of this study was to examine exercise tolerance in patients with COPD from measurements of resting pulmonary function parameters. METHODS: A total of 57 COPD patients were administered the pulmonary function test (PFT) and cardiopulmonary exercise test. The results were analyzed and essentially linear relationships emerged when each subject's VO2 peak was plotted against his individual PFT parameters. Those significant contributors were then introduced in a stepwise multiple regression analysis to determine the best predictor of the VO2 peak. RESULTS: Stepwise multiple regressions in variables revealed that peak oxygen consumption (VO2 peak) was predicted best by the following equation: VO2 peak=(maximum voluntary ventilation x 0.024)+(forced mid-expiratory flow x 0.47)+(body surface area x 0.988)-0.913 (r=0.90; r2=0.81 SE=0.29 L/min). CONCLUSION: We conclude that exercise capacity was predicted from measurements of resting pulmonary function parameters with excellent accuracy in the COPD patient.

An unusual presentation of tuberculosis.

We describe a case of a male with no symptoms and normal chest X ray, diagnosed with TB. The chest computed tomography revealed a cavity formation on the upper left lobe.

Drug-resistance of Mycobacterium tuberculosis in Patras, Greece.

BACKGROUND: The global distribution of drug resistant tuberculosis reflects the quality of tuberculosis control worldwide and is still a major public health-problem. To our knowledge, no data exists in literature about resistance to anti-tuberculosis drugs in Greece. METHODS: The aim of this study was to determine the prevalence of resistance to the main anti-tuberculosis drugs in newly and previously treated tuberculosis patients, in the region of Patras, in Greece and to evaluate the contribution of foreign-born and human immunodeficiency virus (HIV) positive cases to drug resistance. RESULTS: A total of 207 initial isolates of Mycobacterium tuberculosis were analysed. All clinical specimens submitted for cultural diagnosis were collected before chemotherapy commenced. The age of the patients varied from 16 to 78 years old. Sex distribution was 144 males (69.56%) and 63 females (30.43%). Nineteen patients (9.17%) were recent immigrants in Greece. All the patients were HIV-negative. One hundred-one initial isolates of Mycobacterium tuberculosis (48.79%) were susceptible to all first-line anti-tuberculosis drugs, isoniazid, streptomycin, ethambutol and rifampicin. The prevalence of primary mono- and poly-drug resistance was lower (isoniazid 5.79%, streptomycin 4.34%, ethambutol 1.93%, isoniazid + streptomycin 3.38%, isoniazid + ethambutol 0.9%, isoniazid + streptomycin + ethambutol 1.44%) when compared with the prevalence of secondary (acquired) mono- and poly-drug resistance (isoniazid 8.69%, streptomycin 10.14%, ethambutol 3.86%, isoniazid + streptomycin 3.86%, isoniazid + rifampicin 1.44%, isoniazid + streptomycin + ethambutol 4.83%, isoniazid + streptomycin + rifampicin 1.44%). No primary mono - resistance to rifampicin, or primary multi - drug resistance (defined here as Mycobacterium tuberculosis resistant to at least isoniazid and rifampicin) were observed. CONCLUSIONS: According to our data, overall drug resistance was high. Fortunately, multi-drug resistance tuberculosis was not common but special efforts are needed to monitor the prevalence of drug resistance and to ensure adequate treatment in all the population.

Recurrent catamenial hemothorax.

Endometriosis is a common cause of chronic pelvic pain and infertility affecting women of reproductive age, but the disease in rare conditions may be extragenital so may be present with a variety of symptoms. This is a report of an unusual case of pelvic endometriosis that presented with a recurrent hemothorax.

Increased levels of albumin in bronchial washing fluid of patients with bronchial carcinoma. Could albumin be considered as a tumor marker?

BACKGROUND: The aim of this study was to evaluate the significance of albumin in bronchial washing fluid (BWF) and its relationship to three tumor markers (CEA, CA 19-9 and NSE). METHODS: Serum and BWF samples were collected in a group of 60 patients. Albumin and tumor markers in the BWF and serum of three groups: a control group (CG), a chronic bronchitis group (CBG) and a lung cancer group (CaG), were analyzed in a prospective cross-sectional study. The diagnostic yields of the tests in each environment (serum and BWF) were evaluated by using as cutoff points the values of the corresponding 90th percentile of CG and CBG taken together. RESULTS: A significant difference in albumin level (p < 0.001) was noted in the BWF of patients with cancer compared with the other two groups. In addition, a significant difference in CEA level (p < 0.001) was observed in the serum of cancer patients compared with the other two groups. The cutoff values for CEA in serum and albumin in BWF were 2.20 ng/mL and 2.00 g/dL, respectively. The areas under the corresponding ROC curves were 93% and 97%. Combination of CEA-serum and albumin-BWF by logistic regression analysis increased their diagnostic value. CONCLUSION: Measurement of albumin levels in BWF could be a useful additional diagnostic tool to differentiate malignant from non-malignant lung diseases. Moreover, the combined measurement of CEA in serum and albumin in BWF could be of aid in the follow-up of lung cancer patients.

Can high resolution computed tomography predict lung function in patients with chronic obstructive pulmonary disease?

High-resolution computed tomography (HRCT) is a useful method for quantifying the extent of emphysema. Few reports have mentioned the relationships between HRCT scans and pulmonary function tests in chronic obstructive pulmonary disease (COPD). For diagnosis, COPD requires chronic airflow limitation and emphysema and/or chronic bronchitis. We examined 20 who were previous smokers with middle to moderate COPD. All were normocapnic with mean arterial oxygen pressure (PaO2) 77,52 +/- 16,789 mmHg. Forced spirometry, somatic plethysmography and cardiopulmonary exercise test were performed in each patient. HRCT was performed in both full inspiration and full expiration at three levels through the upper (at the aortic arch), lower (2 cm above the diaphragm), and middle lung (midpoint between upper and lower) levels. During expiration all pulmonary function parameters correlated with the HRCT grade in the middle right and left part of the lungs. The middle right part of the lung during expiration correlated statistically significant with MVV (r = -0.681, p = 0.001), forced vital capacity (FVC) (r = -0.477, p = 0.027), forced expiratory volumein 1 sec (FEV1) (r = -0.632, p = 0.002), resistance (r = 0.674, p = 0.001), residual volume (RV) (r = 0.733, p = 0.001), total lung capacity (TLC) (r = 0.696, p = 0.001), functional residual capacity (FRC) (r = 0.752, p = 0.001) and peak oxygen consumption during exercise (VO2) (r = -0.493, p = 0.023). The middle left part of the lung during expiration correlated statistically significant with MVV (r = -0.673, p = 0.001), FVC (r = -0.493, p = 0.027), FEV1 (r = -0.629, p = 0.003), resistance (r = 0.593,p = 0.005), RV (r = 0.601, p = 0.005), TLC (r = 0.546, p = 0.012), FRC (r = 0.594, p = 0.006) and peak VO2 (r = -0.525, p = 0.015). Forced expiratory volume in 1 sec (FEV1), which is a well-established measure of airflow obstruction, correlated with the HRCT grade (1) in the middle left part of the lung during inspiration (r = -0.468, p = 0.035) and during expiration (r = - 0.629, p = 0.003) (2) in the lower right lung during inspiration (r = -0.567, p = 0.007) and during expiration (r = -0.558, p = 0.008) (3) in the lower left lung during inspiration (r = -0.542, p = 0.011) and during expiration (r = -0.558, p = 0.008) (4) in the upper right lung during expiration (r = -0.469, p = 0.037) (5) in the upper left lung during expiration (r = -0.463, p = 0.035) and (6) in the middle right lung during expiration (r = -0.632, p = 0.002). According to our results HRCT was a valuable tool for evaluating the severity of COPD--especially the middle right and left part of the lungs, during expiration--and correlated well with pulmonary function tests.

Abnormal origin of the left common carotid artery by innominate artery: a case of enlargement mediastinum.

An abnormal origin of the left common carotid artery is rare. A 71-year-old male was presented with a cough, fever and dyspnea. A chest radiography revealed a widening of the superior mediastinum. An abnormal origin of the left common carotid artery by innominate artery was diagnosed by angiography.