Modulation of MMP-2 and MMP-9 in Churg-Strauss syndrome respiratory mucosa: potential monitoring parameters.

Churg-Strauss (CSS) syndrome is rare and of unknown etiology. It is associated with vasculitis, blood eosinophilia and granulomatosis, and affects multiple organs and systems at various stages of the disease. Specific diagnostic and monitoring tests are not yet available. This study aims to assess the changes in MMP-2 and MMP-9 along with the histopathological alterations in two cases of CSS, as possible potential diagnostic and monitoring criteria. Two adult male patients were diagnosed with CSS in the otorhinolaryngology clinic in the University of Palermo, based on multiple clinical and histopathologic criteria. Biopsies of respiratory mucosa were taken after the consent of the patients, processed for routine histopathology and immunohistochemistry as well as quantitative polymerase chain reaction (qPCR). Similar biopsies were also taken from a non- CSS patient. The Assessment of MMP-2 and MMP-9 was performed using both immunohistochemistry and qPCR techniques. Histopathological alterations in the respiratory mucosa were consistent with vasculitis and granulomatous tissue formation, in addition to inflammatory cell infiltration with abundance of eosinophils. Immunohistochemistry assay performed on the samples derived from the two CSS patients showed a relative and remarkable increase of both MMP-2 and MMP-9 compared to controls. Such an increase was consistent with the qPCR results which depicted a significant increase between 20 and 30% for both MMP-2 and MMP-9, respectively. Since the secretion of MMPs is an essential step in angiogenesis, could these enzymatic factors be used as parameters to diagnose or monitor the evolution of CSS? The small number of samples analyzed in this study does not allow us to suggest a general statement correlating the increase in expression of MMP-2 and MMP-9 to the appearance or evolution of vasculitis; it is only speculative.

Orthodontic stress Bcl-2 modulation and human odontoblast survival.

This study assessed the effect of orthodontic traction on Bcl-2 expression and apoptosis in human dental pulp. It also explored, in absence of noxious stimuli the regeneration of odontoblasts during the entire life of the tooth. Twenty young patients, with Class II malocclusion and severe to moderate crowding, were referred for orthodontic assessment. Whole pulps were removed. Half the pulps were fixed, paraffin-embedded and processed for histology and immunohistochemistry using anti Bcl-2, Caspase 9 cleaved and Caspase 9 not cleaved antibodies. The rest of the samples, both orthodontically treated and not treated dental pulps, were immediately frozen at -80ºC after the extraction and quantitative PCR was performed. Histology showed alterations in pulp microanatomy after 8 months of treatment. Immunohistochemistry depicted a decreasing expression of Bcl-2 in dental pulp over time in the non-treated while a very weak to absent Bcl-2 expression was detected in the orthodontically treated tissues. Active and non-active forms of Caspases, were expressed in both groups of dental pulp, however staining for the non active form was stronger than the corresponding cleaved form in all samples. The increased expression was detected mainly at nuclear level. Real time qPCR results correlated with those of immunohistochemistry and exhibited a decreasing expression of Bcl-2 in the treated samples. Orthodontic traction may inhibit the expression of Bcl-2, favoring the onset of apoptosis and leading us to conclude that the physical stress in the absence of noxious stimuli might make odontoblasts regeneration less likely.

Prevalence of IL-1ß +3954 and IL-1α-889 polymorphisms in the Lebanese population and its association with the severity of adult chronic periodontitis.

Pro-inflammatory cytokines, i.e., IL-1 mediate the inflammatory response and are genetically regulated in periodontal diseases. Strong association was found between the composite genotype allele 2 of IL-1ß+3954 and IL-1α-889 and severe chronic periodontitis. The aim of this study is to determine the prevalence of IL-1ß+3954 and IL-1α-889 polymorphism in a group of Lebanese individuals of homogeneous ethnicity and the possible association between genotype positive individuals and the severity of periodontal disease. One hundred and fifty-seven patients aged 53.29±13.13 years participated in the study. Subjects were classified as follows: 1) healthy subjects with no attachment loss >1mm and no clinical signs of gingival or periodontal inflammation; 2) diseased subjects with mild periodontitis (less than 15‰ of global periodontal bone loss); 3) subjects with moderate periodontitis (less than 4 interproximal sites with bone loss = or >50 percent and mean bone loss between 15 and 30%); 4) subjects with severe periodontitis (more than 7 interproximal sites with >50% bone loss and mean bone loss >35‰). Blood samples were taken and analyzed for polymorphism in the IL-1α gene at position +4845 and in the IL-1beta gene at position +3953. Statistical analysis was performed using chi-square test, Fisher Exact test, and ANOVA followed by Bonferroni multiple comparisons. The prevalence of genotype-positive subjects was 52.3‰ in the healthy control group and 42 ‰ in the diseased group. Positive genotype heterozygous of allele 1 and 2 for IL-1ß+3954 and IL-1α-889 did not represent in this study a major risk for chronic periodontitis (p=0.590). Only subjects homozygous for allele2 of the IL-1ß+3954 and IL-1α-889 were significantly more at risk for severe periodontitis with OR of 51.42.

Rat model of burn wound healing: effect of Botox.

Animal models of burn play a crucial role in studying the mechanisms of burn wound progression and the factors that regulate various stages of healing. In this study, using a rat model, we assessed the effect of Botox in the healing process through parameters like transepidermal water loss (TEWL), histological alterations, transforming growth factor beta (TGF-beta1) and tumor necrosis factor alpha (TNF-alpha). Fifty Sprague-Dawley rats were inflicted with 5 cm2 second degree burn and divided into 2 groups; one group was injected intralesionally with Botox and the other with saline. Daily observation and transepidermal water loss measurement were performed. Biopsies were taken on days 0, 3, 8, 14, and 28 for histology and polymerase chain reaction, testing TGF-beta and TNF-alpha. The results showed no significant difference in TEWL except for slightly better preservation of moisture with Botox. Histology revealed relatively better and faster regeneration with Botox, delayed lower grade inflammation, and increase in fibroblasts. TNF-alpha had an acute increase of 21-fold then tapered down while TGF-beta levels increased on day 3 after TNF-alpha, peaked on day 8 and then started to decrease until complete healing. Botox improved the healing process and the cosmetic appearance of burn scar.

Enteropathogenic e.coli sustains iodoacetamide-induced ulcerative colitis-like colitis in rats: modulation of IL-1ß, IL-6, TNF-α, COX-2, and apoptosisi.

Pathogenic or non-pathogenic bacteria from flora may play a key role in inflammatory bowel disease (IBD) pathogenesis. However, a specific infectious agent causing IBD has not been identified. This study assessed the impact of enteropathogenic E. coli (EPEC) on the modulation of IL-1beta, IL-6, TNF- alpha, COX-2, BAX and Bcl-2 expression, in sustaining inflammation of a rat colitis model. Two hundred male Sprague-Dawley rats (4 groups) were inoculated weekly or bi-weekly for 70 days, with 1 percent methylcellulose (MC), (b) 6 percent iodoacetamide (IA) in 1 percent MC, (c) 4x108 CFU of EPEC, and (d) IA+EPEC. After a month, treatment was stopped in half of the animals in each group. IL-1beta, IL-6, TNF-alpha, COX-2, BAX and Bcl-2 expression were measured in colonic mucosa scrapings. IL-1beta, IL-6, TNF-alpha, and COX-2 were significantly increased in colonic mucosa of the IA+EPEC group and to a lesser but significant level in the IA group compared to controls, or EPEC alone, both in continued and discontinued treatment groups. Additionally, the BAX/Bcl-2 ratio decreased, indicating less apoptosis in the IA+EPEC group which exhibited more necrosis. These effects increased with experiment duration. This work provides new arguments favouring the role of bacteria in IBD pathogenesis.

Cytology and clinical spectrum of sexually transmitted infections in Lebanese women as revealed by Pap smear: a cross-sectional study from 2002-2006.

To study the cytology profile of cervical smears and the respective prevalence and incidence of certain cervico-vaginal infections detectable by routine Pap smear cytology in Lebanese women from 2002 to 2006. Pap smear cytology results were compiled from the archives of the Institut National de Pathologie for the period extending from 2002 until 2006. This study covered 118,230 cervical specimens obtained from Lebanese women attending clinics and hospitals in all the five districts of Lebanon; prevalence and incidence rates for infections detectable by routine Pap smear examination were determined. A rise in prevalence of these infections by 2.1 percent (2,555) from 2002 to 2006 was revealed. A doubling of Pap smears showing HPV-associated changes was detected (1.4 percent in 2002 to 2.9 percent in 2006), and a simultaneous almost 7-fold increase of Atypical Squamous Cells of Undetermined Significance (ASCUS) cytology was detected during this period; the rise in ASCUS cytology was age-dependent. Moreover, a 60 percent increase in prevalence of bacterial vaginosis (2.3 percent in 2002 to 3.7 percent in 2006) and a more than 3-fold decline in Trichomonas vaginalis infection (1 percent in 2002 to 0.3 percent in 2006) were also noted in this population during this period. An increase in the prevalence and incidence of cervico-vaginal infections detectable by Pap smear cytology in Lebanese women was revealed from 2002 to 2006. Such changes could point to recent modifications of sexual and health behaviours in the Lebanese community.

Review and update on the use of the microanastomotic coupler device for arterial anastomosis in free tissue transfer.

While a simple interrupted sutured anastomosis remains the gold standard in microsurgery, the introduction of the microanastomotic coupler device (MACD) has decreased procedure time and thrombosis risk, and improved the patency of venous anastomoses. The aim of this review is to update the evidence-based advantages of the MACD on arteries, based on clinical and experimental data, and to compare them to the hand-sewn approach in free flap transfer. All relevant articles that appeared in the PubMed and Medline/Ovid databases during the past three decades were reviewed. After exclusions, 11 studies were retained and discussed. The MACD had a generally shorter arterial anastomosis time, with improved flap survival and reduced ischaemia compared with the hand-sewn approach. The use of the MACD in arterial anastomosis is an efficient and less time-consuming alternative to the hand-sewn technique, provided that the selection of vessels is appropriate and the vessel diameter is large enough to do the anastomosis.

Knowledge, attitudes, beliefs and practices in Lebanon concerning HIV/AIDS, 1996-2004.

This cross-sectional study, aimed at evaluating the impact of HIV prevention interventions in Lebanon since 1996, was performed between January 2004 and July 2004 on 3200 Lebanese aged 15-49 years. Of the sexually active respondents, 13.0% of men and 2.6% of women had regular partners other than the spouse but only 25.0% used a condom in their last sexual intercourse. However, 16.8% had sex with casual partners and 71.7% of those used a condom. Knowledge about preventive practices against HIV/AIDS has regressed since 1996, 85.7% compared to 94.9%. Self reports of symptoms suggestive of sexually transmitted disease were 9.1% compared to 5.6% in 1996.

The prevalence of hypertension and its association with other cardiovascular disease risk factors in a representative sample of the Lebanese population.

Hypertension is a well-known major risk factor for cardiovascular diseases. This study has been designed to assess the prevalence of hypertension, its co-occurrence with other cardiovascular risk factors and its association with cardiovascular diseases in a representative sample of Lebanese population. A pretested questionnaire was administered to 2125 adults aged equal or above 30 years old from all regions in Lebanon and proportionate with the respective population density. Data revealed that 23.1% of the respondents admitted being hypertensive, with no statistically significant difference between male and female patients. Prevalence of hypertension increased with age (P < 0.01) and hypertension tended to occur more in the less educated and the unemployed (P < 0.01). Only 45.7% of hypertensive patients followed a low fat diet while 14.7% exercised daily. The prevalence of hypertension increased significantly with an increase in body mass index particularly in female patients. Among the hypertensive respondents, 23.9% were diabetic, 38.1% were hyperlipidaemic and 9.1% were both diabetic and hyperlipidaemic. Co-occurrence of hypertension with diabetes, hyperlipidaemia or both was more common in female patients compared to male patients. In addition, stroke, myocardial infarction (MI) and atherothrombosis occurred in hypertensive respondents at rates of 2.4, 4.3 and 9.5%, respectively. Hypertensive female patients reported more stroke and atherothrombotic episodes compared to male patients, whereas hypertensive male patients had more MIs. Based on these results it is very important to address the issue of lifestyle modification for the prevention and treatment of hypertension and awareness campaigns should stress the fact that cardiovascular diseases are not only restricted to men.

Adherence to universal precautions among laboratory personnel in Lebanon.

To evaluate the present situation and plan future directions with regard to implementation of universal precautions in laboratories testing blood samples, we carried out a national cross sectional study in 2003 on a representative sample of laboratories in Lebanon. We compared the results with those of a 1993 study. We found that the education profile of staff had improved, being now more specialized in laboratory science. The discrepancies between what technicians knew, believed in and practised and what was observed in the field improved to some extent in most variables. Disposal of needles and syringes had improved greatly but disposal of blood-contaminated material had not. Given the risks of improper practice, a policy of universal precautions is essential and regular training should be carried out so that staff know and practise the universal precautions and correct laboratory procedures.

Distribution of substance P positive cells and nerve fibers in the rat thymus.

The immune and nervous systems communicate through an array of signalling molecules which includes substance P. This work investigates the anatomical relationship between substance P nerve fibers, receptors, and substance P positive cells in the thymus. Thymuses from rats were frozen or paraformaldehyde fixed. In vitro autoradiography was used to map the distribution of SP receptors. Immunostaining was used to localize SP positive cells and nerve fibers by transmitted light and confocal microscopy. SP receptors exhibited a broader distribution than previously reported, being present throughout the organ with a preferential concentration in the cortico-medullary zone. While SP fibers were frequently associated with the blood vasculature, they were also present throughout the organ independent of blood vessels and were most prominent in the cortico-medullary zone. SP positive cells followed a similar pattern of distribution as the SP fibers and were present as single cells or aggregates of SP positive cells. Confocal microscopy revealed close spatial contact between the SP positive nerve fibers and the SP positive thymic cells. The close spatial relationship between the SP positive thymic cells and SP positive nerve fibers supports the concept of a structural-functional unit between SP nerve fibers and their potential receptor-bearing target cells in the thymus.

Prolactin receptor localization to the area postrema.

The area postrema, which lacks a blood-brain barrier, was examined for the presence of prolactin receptors, which would render it a potential site for vascular prolactin to directly interact with neuronal elements. Using an in vitro autoradiographic technique, frozen sections of New Zealand white rabbit medulla were incubated with radiolabelled ovine prolactin alone (total binding) or radiolabelled ovine prolactin in the presence of excess unlabelled ovine prolactin (non-specific binding). The specificity of the binding was also assessed using excess unlabelled human prolactin or ovine LH. While excess unlabelled ovine and human prolactin caused a statistically significant reduction in radio labeled prolactin binding, unlabelled LH was without effect. Results reveal the presence of specific prolactin binding sites within the area postrema, a previously unknown prolactin target area of the CNS.

Management of hemorrhoids by rubber band ligation.

This study discusses the management of hemorrhoids by rubber band ligation. It reviews 233 rubber band ligation procedures in 148 patients with symptomatic hemorrhoids performed over a 5-year period. 72% of the patients underwent a single rubber band ligation, 22% a double band ligation and 6% a triple band ligation in one session without anesthesia. A follow-up of 18 +/- 5 months was carried out. Only three patients required hospitalization for complications. One patient was admitted for bleeding and recovered with clot evacuation, bed rest and blood transfusion. Two patients developed a band related abscess that resolved with drainage. Twenty patients (13.5%) reported pain associated with the procedure. Four patients needed subsequent surgical hemorrhoidectomy. Long-term results were good in 81.2% of the patients. This report agrees with previously published series showing the efficacy and cost containment of hemorrhoidal ligation.

Modulation of some human mononuclear cells activities by procaine.

The in vitro effects of a local anesthetic, a membrane active drug, procaine, on some functional activities of circulating human lymphocytes and monocytes were studied. Procaine inhibited spontaneous E-rosette formation between T-lymphocytes and sheep erythrocytes and EAC-rosettes with B-lymphocytes. In addition, procaine inhibited both the phagocytosis of latex particles by normal monocytes and the proliferation of lymphocytes in an allogeneic mixed leukocyte culture. Morphologically the procaine-treated cells exhibited a relative increase in the size of the cytoplasmic rim around their nuclei. The results indicated that procaine might be considered as a non-specific immunoregulator, modulating to some extent the functional expression of human peripheral blood mononuclear cells activities.

A comparative study of morphological changes in spontaneously hypertensive rats and normotensive Wistar Kyoto rats treated with an angiotensin-converting enzyme inhibitor or a calcium-channel blocker.

It is not clear whether some pathological changes in hypertension are directly pressure-dependent, or hormonally induced, or both. The aortic arch has apparently never before been studied for those changes. The aim of this study was to compare the effects of controlling angiotensin II (Ang II) and/or blood pressure (BP), directly at the inception of hypertension, on the aortic arch, the left ventricle of the heart (LV), and the kidneys of spontaneously hypertensive rats (SHRs) and normotensive Wistar Kyoto (WKY) rats. An angiotensin-converting enzyme inhibitor (ACEI, enalapril) and a calcium-channel blocker (nifedipine) were used for 21 weeks. After treatment, rats were assessed for arterial plasma renin activity (PRA). The LV, aortic arch, and kidneys were then excised for the determination of organ and tissue weight in some of the animals, while in others the aortic arch was fixed in situ and processed for microscopic analysis. Both enalapril and nifedipine levelled BP in the SHRs to almost normal values. Enalapril was able to prevent the increase in LV and kidney weights (p=0.04 wet, p<0.001 dry; p<0.001 wet and dry, respectively) and the increase in the weight of the aortic arch and in the thickness of its media (p<0.001 wet and dry; p<0.001, respectively) seen in untreated SHRs. This was associated with a larger lumen diameter (p<0.001) and a lower media to lumen ratio (p=0.01). In contrast, nifedipine did not prevent any of the changes described. Neither nifedipine nor enalapril treatment had any effects on PRA in either rat strain. Our results support previous observations that BP is not the only factor causing some of the pathological changes in hypertension; tissue Ang II level may also play a major role.

Cysteamine-induced duodenal ulcer: effects on calcium absorption across rat duodenum.

The effect of cysteamine-induced duodenal ulcers on calcium transport across rat duodenum was investigated. Intracellular calcium accumulation measured after 24 hr and 3 days of cysteamine injection showed a significant increase (P less than 0.001) in the duodenal strips isolated after 3 days with no change noticed in those isolated after 24 hr, although the morphological changes in both were very similar. The relationship between increasing calcium concentration in the incubation medium and intracellular calcium concentration is a saturable process that conforms to the Michaelis-Menten type of kinetics. The average maximal flux (Vmax) increased from 8.93 nmole/hr-gdw in normal to 12.5 nmole/hr-gdw in 3-day-ulcerated rats, with no apparent change in the Michaelis constant (Kt) (0.8 mM). Unidirectional influx of calcium across the mucosal membrane was significantly increased (P less than 0.001) in 3-day-ulcerated duodenum suggesting that the increase in calcium transport could be due to the activation of the active step at the mucosal border.