PCR-Internal Transcribed Spacer (ITS) genes sequencing and phylogenetic analysis of clinical and environmental Aspergillus species associated with HIV-TB co infected patients in a hospital in Abeokuta, southwestern Nigeria.

Background: Aspergillosis has been identified as one of the hospital acquired infections but the contribution of water and inhouse air as possible sources of Aspergillus infection in immunocompromised individuals like HIV-TB patients have not been studied in any hospital setting in Nigeria. Objective: To identify and investigate genetic relationship between clinical and environmental Aspergillus species associated with HIV-TB co infected patients. Methods: DNA extraction, purification, amplification and sequencing of Internal Transcribed Spacer (ITS) genes were performed using standard protocols. Similarity search using BLAST on NCBI was used for species identification and MEGA 5.0 was used for phylogenetic analysis. Results: Analyses of sequenced ITS genes of selected fourteen (14) Aspergillus isolates identified in the GenBank database revealed Aspergillus niger (28.57%), Aspergillus tubingensis (7.14%), Aspergillus flavus (7.14%) and Aspergillus fumigatus (57.14%). Aspergillus in sputum of HIV patients were Aspergillus niger, A. fumigatus, A. tubingensis and A. flavus. Also, A. niger and A. fumigatus were identified from water and open-air. Phylogenetic analysis of sequences yielded genetic relatedness between clinical and environmental isolates. Conclusion: Water and air in health care settings in Nigeria are important sources of Aspergillus sp. for HIV-TB patients.

Stapedotomy and its effect on hearing – our experience with 54 cases.

Introduction: This study reviews the cases of stapedotomy and evaluates its effectiveness at improving hearing loss in patients with otosclerosis. Materials and methods: Retrospective review of patients’ records who had clinical and audiometric diagnosis of otosclerosis from January to December 2012. Results: A total of 54 stapedotomy surgeries (52 unilateral and 1 bilateral surgeries) were received. Average post-operative ABG for 0.5, 1, 2 and 4 KHz showed that 61.1% had complete closure of ABG (ABG < 10 dB) compared with 1.6% preoperatively and 85.2% had closure of ABG to within 20dB compared with 4.7% preoperatively (t – 13.89, p = 0.000). More than 94% had hearing improvement and 81.5% had ABG closure greater than 10 dB postoperatively (mean gain 23.38 ± 12.37, t = 13.89, p = 0.000). A total of 13% complications were recorded with TM perforation (5.6%) being the commonest complication. Conclusion: Stapedotomy is an effective surgical procedure for the treatment of otosclerosis which leads to improvement in patients’ quality of life. A favorable hearing outcome can be obtained by the combination of experienced hands with minimal surgical trauma and appropriate surgical technique.

COMMUNITY KNOWLEDGE, ATTITUDE AND PRACTICES (KAP) ON FLUOROSIS AND ITS MITIGATION IN ENDEMIC AREAS OF ETHIOPIA

Optimum fluoride intake plays an essential role in the prevention of dental caries while fluoride consumption above recommended level interferes with the normal formation of tooth enamel and bones and may increase risk of dental and skeletal fluorosis. The knowledge and practices of endemic communities on etiology of fluorosis will help in its mitigation and prevention. The objective of this study was to investigate the knowledge, attitude and practices of endemic community on fluoride contamination, fluorosis and prevention practices in order to devise coordinated and targeted prevention mechanisms. Focus group discussions (FGD) and key-informant interview were conducted in three dietary areas to collect knowledge, attitude and practices (KAP) of the endemic community in July 2013.The results indicated that health consequences of fluoride contaminated water are fairly understood. None of the discussants mentioned the word “fluoride”. The knowledge and perception of the community on fluoride ingestion is poor. Health extension workers (HEWs) did not teach about fluoride and related health consequences. Dental fluorosis was reported to start at early ages and not commonly perceived as a major problem. However, adolescents worried and felt that they might be singled out when going to other areas. Older people have a skeletal fluorosis, which interferes with their day to day activities. In severely affected people, the teeth were weak and fragile and thus create difficulty in chewing hard foods like unfermented dry flat bread, sugar cane and toasted grains. People prefer rain water rather than water from borehole because of the inconvenient taste of the latter. The endemic communities have no sufficient knowledge and skills on potential sources of fluoride intake, the debilitating effect of high fluoride ingestion, and preventive and mitigatory measures to reduce fluoride intake. The effect of fluoride contamination and mitigatory methods should get sufficient attention by the community, health workers and concerned governmental bodies. The trend of harvesting and using rain water should be encouraged as it reduces fluoride intake. Future studies should focus on information communication on possible fluoride risks, intervention and evaluation studies on defluoridation, rain water harvesting and mitigatory techniques.

The identification and differentiation of the Candida parapsilosis complex species by polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region of the rDNA

Currently, it is accepted that there are three species that were formerly grouped under Candida parapsilosis : C. parapsilosis sensu stricto, Candida orthopsilosis , and Candida metapsilosis . In fact, the antifungal susceptibility profiles and distinct virulence attributes demonstrate the differences in these nosocomial pathogens. An accurate, fast, and economical identification of fungal species has been the main goal in mycology. In the present study, we searched sequences that were available in the GenBank database in order to identify the complete sequence for the internal transcribed spacer (ITS)1-5.8S-ITS2 region, which is comprised of the forward and reverse primers ITS1 and ITS4. Subsequently, an in silico polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to differentiate the C. parapsilosis complex species. Ninety-eight clinical isolates from patients with fungaemia were submitted for analysis, where 59 isolates were identified as C. parapsilosis sensu stricto, 37 were identified as C. orthopsilosis, and two were identified as C. metapsilosis. PCR-RFLP quickly and accurately identified C. parapsilosis complex species, making this method an alternative and routine identification system for use in clinical mycology laboratories.

Expression of TIMP-2 in HPV-16 infected oral squamous cell carcinoma in patients in Iraq

Aim: To determine the expression of tissue inhibitors of metalloproteinases (TIMP-2) in oral squamous cell carcinoma (OSCC) and the difference in its expression level between positive and negative HPV-16 (human papilloma virus- 16) OSCC patients. Methods: This study was conducted on 33 biopsies obtained from patients with OSCC and 10 normal oral mucosa as controls. In situ hybridization (ISH) was used to investigate the presence of HPV-16, while immunohistochemistry (IHC) was used to estimate the expression level of TIMP-2. Results: The TIMP-2 was expressed in 27 (81.8%) of OSCC sections with no significant difference between its expression level in HPV-16 positive and HPV-16 negative OSCC cases (p=0.058). TIMP-2 was found to be highly expressed in OSCC sections, and the presence of HPV was not related to its overexpression. Conclusions: The percentage of samples that appeared to accommodate detectable HPV-16 was high, but no significant difference was observed in relation to TIMP-2 expression level. Future studies with a larger number of patients are highly recommended to address the possible association between TIMp-2 and OSCC positive HPV-16.

Prevalence of Mental Health Problems in Children and Its Associated Socio-Familial Factors in Urban Population of Semnan, Iran (2012)

Background: The World Health Organization (WHO) defines mental health as “a state of well-being in which every individual realizes own potential, can cope with the normal pressures of life, is able to work effectively, and can make a contribution to community”. Objectives: Mental Health Problems (MHP) is a great concern for all societies in terms of its burden and impact. This survey screened MHP and its impact in an Iranian urban population aged 6 - 12 years old, and explored its associated socio-familial factors. Patients and Methods: The survey was conducted in the elementary schools of Semnan, using random cluster sampling. Collection and analysis of data was performed using the parent version of the “Strengths and Difficulties Questionnaire (SDQ)” and survey commands of Stata-nine, taking into account cluster effect and population weights. Associations were assessed by fitting simple and multiple logistic regression models. P < 0.05 was considered significant. Results: With regard to the SDQ total score, 19.3% (95% CI: 8.6, 30.1) scored above the normal threshold (9.6% abnormal, 9.7% borderline). The frequency of problems ranged between 16.1% (peer problems) and 8.4% (emotional symptoms), and in all subscales boys were affected more than girls. The impact score was abnormal in 68.4% of all children, and was greater in girls than in boys. “A previously diagnosed mental health disorder” (OR = 11.11, 95% CI: 5.55, 25.00), “male gender” (OR = 1.43, 95% CI: 1.10, 1.87 and “less time spent with the child by father” (OR = 1.61, 95% CI: 1.20, 2.17) were significantly associated with an abnormal SDQ. Conclusions: The high rate of MHP in 6 - 12 year-old children and the lack of any significant correlation with their age, underpins the importance of early screening for MHP in schools, with particular focus on high risk groups.

Th1/Th2 Cytokine Profile and Its Diagnostic Value in Mycoplasma pneumoniae Pneumonia

Background: The levels of Th1/Th2 cytokine can alter in pathogenic infection in children with pneumonia. Objectives: To evaluate Th1/Th2 cytokine profile and its diagnostic value in M. pneumoniae pneumonia in children. Patients and Methods: Children with M. pneumoniae mono-infection and 30 healthy children were tested with cytokines assay. We used real time PCR to detect M. pneumoniae in children with pneumonia. Results: M. pneumoniae test was positive in 2188 (16.62%) out of 13161 pneumonia children. Children aged 5 - 9 years had the highest rate and summer was a season with high rate of M. pneumoniae incidence in Zhejiang province. During the course of study, in 526 pneumonia children with M. pneumoniae mono-infection and 30 healthy children cytokines assay was performed. IL-2 level of M. pneumoniae pneumonia children was lower than that of healthy children (median levels, pg/mL: IL-2: 3.2 vs. 5.7, P = 0.00), while IL-4, IL-10 and IFN-γ were higher than in healthy children (median levels, pg/mL: IL-4: 3.2 vs. 1.5, P = 0.00; IL-10: 5.6 vs. 2.5, P = 0.001; IFN-γ: 20.4 vs. 4.8, P = 0.001). Conclusions: IL-2 decreases and IL-4, IL-10 and IFN-γ increase in children with M. pneumoniae pneumonia, which has a promising prospect in diagnosis of this disease in clinical practice.