Evaluation of cryoinjury of spermatozoa after slow (programmed biological freezer) or rapid (liquid nitrogen vapour) freeze-thawing techniques

Purpose: This study was initiated to determine the negative effect (cryodamage) on human spermatozoa after freeze–thawing and to find out whether freezing of spermatozoa with a computerized biological freezer is more advantageous than freezing above static liquid nitrogen vapour with regard to spermatozoa vitality, chromatin normality, morphology, and membrane integrity. Methods: Forty-four semen samples were obtained from patients attending andrology laboratory, and each sample was divided into two aliquots. One aliquot was frozen using static liquid nitrogen vapour (G.II) and the second with a computerized biological freezer (G.III). Acridine orange was used for assessment of chromatin cryoinjury, whereas the morphology was evaluated according to WHO criteria. Hypo-osmotic swelling test was used to identify membrane integrity and eosin–nigrosin staining was used to determine the vitality of spermatozoa. Results: The mean percentage of normally condensed chromatin in the native semen sample (G.I) decreased significantly (p < .001) after freeze–thawing by using either liquid nitrogen vapour (G.II), or a biological freezer (G.III), which was significantly higher (p < .001) after freezing with liquid nitrogen vapour than after freezing with the biological programmed freezer. Morphologically normal spermatozoa decreased significantly (p < .001) in both freezing methods in comparison to the native semen samples. In addition, membrane integrity of spermatozoa (HOS-test positive) was significantly lower (p < .001) after the freeze–thawing procedure in G.II and G.III compared to G.I. In both these parameters the deterioration was similar among the two freezing procedures. Finally the mean percentage of live spermatozoa decreased significantly (p < .001) in both freezing techniques in relation to the mean value in the neat semen samples. Conclusions: Freeze–thawing procedure has a detrimental effect on chromatin, morphology, membrane integrity, and vitality of human spermatozoa not only by freezing above static liquid nitrogen vapour but even by using a computerized biological freezer. However, the chromatin deterioration rates are significantly higher by freezing above static liquid nitrogen vapour in comparison to freezing with a programmed biological freezer. Therefore, we recommend the use of this technique for freezing semen especially when ICSI technique is considered as the main therapeutic procedure.     

The role of histamine in human mammary carcinogenesis: H3 and H4 receptors as potential therapeutic targets for breast cancer treatment

There is increasing evidence that describes a histamine role in normal and cancer cell proliferation. To better understand the importance of histamine in breast cancer development, the expression of histamine H3 (H3R) and H4 (H4R) receptors and their association with proliferating cell nuclear antigen (PCNA), histidine decarboxylase (HDC) and histamine content were explored in mammary biopsies. Additionally, we investigated whether H3R and H4R were implicated in the biological responses triggered by histamine in MDA-MB-231 breast cancer cells. The expression levels of H3R, H4R, PCNA, HDC and histamine content were determined by immunohistochemistry in 40 benign and malignant lesions. MDA-MB-231 cells proliferation (clonogenic assay and BrdU incorporation) and cell cycle distribution (flow cytometry) were evaluated upon treatment with histamine, H3R and H4R agonists and antagonists. Apoptosis was determined by Annexin staining and TUNEL assay. Cell migration was assessed by transwell system. Results indicate that H3R was detected in 67% (10/15) of benign lesions and in almost all carcinomas (24/25), being the level of its expression significantly higher in carcinomas (p = 0.0016). The non-tumoral breast tissue surrounding carcinomas revealed a lower H3R expression compared to the tumor cells. Only 13% (2/15) of the benign lesions expressed H4R compared to 44% (11/25) of the carcinomas. Interestingly, H3R expression was correlated in carcinomas with the expression of HDC and PCNA (p < 0.0001), and also histamine content (p = 0.0229). Accordingly, histamine increased MDA-MB-231 cells proliferation and also migration via H3R. In contrast, activation of H4R inhibited proliferation and this effect was associated with an arrest in the G(0)/G(1) phase of the cell cycle and an induction of apoptosis. Present findings demonstrate the presence of H3R and H4R in human mammary tissue and suggest that H3R may be involved in the regulation of breast cancer growth and progression representing a novel molecular target for new therapeutic approach.     

Rationale and methodology for a population-based study of eye diseases in Malay people: The Singapore Malay eye study (SiMES)

PURPOSE: Although there are approximately 200 million people of Malay ethnicity living in Asia, the burden and risk factors of blinding eye diseases in this ethnic group are unknown. This study summarizes the rationale and study design of a population-based study of eye diseases among adult Malays in Singapore. METHODS: A population-based cross-sectional study of Malays was designed in Singapore. The sampling frame consisted of all Malays aged 40-79 living in designated study areas in southwestern Singapore. From a list of 16,069 names provided by the Ministry of Home Affairs, age-stratified random sampling was used to select 5,600 names (1,400 people from each decade of 40-49, 50-59, 60-69, and 70-79 years). The target sample size for this study was 3,150 persons. Selected individuals were invited to a centralized clinic by letters, telephone calls, and home visits. Participants underwent standardized interview and assessment of blood pressure, anthropometry, presenting and best-corrected visual acuity, subjective refraction, ocular biometry, Goldmann tonometry, slit-lamp biomicroscopy, optic disc imaging, digital lens, and retinal photography. Blood and urine samples were collected for biochemical analyses and further stored for future studies. Selected participants also had gonioscopic examination, visual fields test, and assessment of ankle and brachial blood pressure to detect presence of peripheral vascular disease. CONCLUSIONS: This study provides population-based data on the prevalence of and risk factors for age-related eye diseases in people of Malay ethnicity in Singapore. Data from this study allow further understanding of the etiology and impact of eye diseases in this ethnic group.     

Socioeconomic Factors Association with Knowledge and Practice of Breast Self-Examination among Iranian Women.

Background: Breast cancer is the leading type of cancer in women and is one of the most frequent cancers among Iranian women. Delay in diagnosis and treatment of breast cancer diminishes a women's chance of survival. Breast self- examination (BSE) may be effective in early detection. The purpose of this study was to identify the relationship between Iranian women's socioeconomic status and their knowledge and practice of BSE. Methods: Data were from a hospital-based case-control study among women diagnosed with breast cancer. Control subjects were matched to patients on age. 303 breast cancer patients and 303 control women were interviewed. Socioeconomic status and information including knowledge and practices of breast self examination and clinical breast examination were recorded and compared. Results: The mean +/- SD age of cases and controls was 48.2 +/- 9.8 and 50.2 +/- 11.1 (range 24-84 years), respectively. The study revealed that there were significant relationships between education level and knowledge and practices of breast self examination in both cases and controls, increase in usage being observed with the level of education (P<0.05). Conclusion: The findings suggest that the knowledge and practices of women toward breast cancer early detection are inadequate in women with a lower level of education. Mass media cancer education should promote widespread access to information about early detection behavior.     

The effect of dental overbite on eustachian tube dysfunction in Iranian children

OBJECTIVE: To investigate the association between deep dental overbite and eustachian tube dysfunction. METHODS: It was designed as a case-control study. Among hospitalized patients in otolaryngology department at Taleghani Hospital in Tehran, Iran, from January to December 2005, 132 patients between the ages of 2 and 6 years were recruited. Dental overbite, overjet, and occlusal relationships were measured by one observer. Eustachian tube dysfunction was defined as having ventilation tubes with an abnormal tympanometry. In addition, demographic information, medical and social histories were prospectively recorded. Univariate and multivariate logistic regression model were used. RESULTS: In a multivariate model, children with deep bites were 10.6 times more likely to have eustachian tube dysfunction than those without deep bites (P<0.05). Other independent risk factors for eustachian tube dysfunction identified in this model were family history of otitis media, daycare exposure, and non-breast-feeding. CONCLUSIONS: Children with deep dental overbites are at a significantly increased risk for developing eustachian tube dysfunction.     

Proinflammatory high‐density lipoprotein as a biomarker for atherosclerosis in patients with systemic lupus erythematosus and rheumatoid arthritis

Objective

Women with systemic lupus erythematosus (SLE) have a 7–50‐fold increased risk of coronary artery disease (CAD). In the general population, oxidized low‐density lipoprotein (ox‐LDL) increases the risk for CAD. Normal high‐density lipoproteins (HDLs) protect LDL from oxidation; proinflammatory HDLs do not. This study was undertaken to determine whether patients with SLE, who have chronic inflammation that causes oxidative damage, have more proinflammatory HDL and higher levels of ox‐LDL, thus predisposing them to atherosclerosis.

Methods

One hundred fifty‐four women with SLE, 48 women with rheumatoid arthritis (RA), and 72 healthy controls were studied. The ability of the patients' HDL to prevent oxidation of normal LDL was measured. Values >1.0 (the value assigned for LDL oxidation in the absence of HDL) after the addition of HDL indicated proinflammatory HDL. Plasma ox‐LDL levels were measured as the amount of oxidation produced by the patient's LDL after the removal of HDL.

Results

SLE patients had more proinflammatory HDL (mean ± SD score 1.02 ± 0.57, versus 0.68 ± 0.28 in controls [P < 0.0001] and 0.81 ± 0.22 in RA patients [P = 0.001 versus SLE patients]). A higher proportion of SLE patients had proinflammatory HDL: 44.7% of SLE patients versus 4.1% of controls and 20.1% of RA patients had scores >1.0 (P < 0.006 between all groups). Levels of ox‐LDL correlated with levels of proinflammatory HDL (r = 0.37, P < 0.001). SLE patients with CAD had significantly higher proinflammatory HDL scores than patients without CAD (P < 0.001).

Conclusion

HDLs are proinflammatory in a significant proportion of SLE patients and are associated with elevated levels of ox‐LDL. Abnormal HDLs impair the ability to prevent LDL oxidation and may predispose to atherosclerosis.

     

Diagnostic accuracy of activities of daily living in prediction of community-acquired pneumonia outcomes in elderly patients admitted to intensive care units

Background/PurposeMost severity scores can predict severe community acquired pneumonia (CAP) and its associated poor outcomes. The impact of baseline functional status on the prediction of the outcome of CAP remains controversial. Therefore, we aimed to detect the diagnostic accuracy of the baseline function using activities of daily living (ADL) in prediction of mechanical ventilation (MV) and 30-day mortality and to compare its diagnostic accuracy with the CURB-65 and SCAP scores in patients with CAP.MethodsA prospective cohort study was conducted on 65 patients aged ≥60 years presenting with CAP and admitted consecutively to geriatric and chest intensive care units in Ain Shams University Hospitals from October 2011 to June 2012. Patients were subjected to assessment of the severity of CAP using the CURB-65 and SCAP scores, and for baseline function using the ADL score. Follow-up for MV and 30-day mortality was also done.ResultsThe mean age of the patients was 69.9 ± 11.4 years; 40% of patients died, 41.5% were on MV, and 75.4% had ADL score ≤ 3 points. By logistic regression, the ADL score was found to be an independent predictor for mortality and MV in patients with CAP after adjusting for confounding factors. Using ADL score for prediction of mortality and MV was best at cut off ≤ 3, which revealed area under the curve (AUC) = 0.705, 0.679; p = 0.005, 0.015; sensitivity = 100%, 96.3%; specificity = 41% and 39.5%, respectively. Pairwise comparison between AUCs of the ADL score and other scores revealed no significant difference.ConclusionThe ADL score can be an attractive alternative to conventional indices as it is an independent predictor for mortality and MV in patients with CAP.