It is well established that water quality is directly linked to health. In-line chlorination is one technology that can be used in the developing world to potentially inactivate pathogens and improve water quality. The purpose of this study was to determine the effectiveness of the Panamanian Ministry of Health's in-line PVC chlorinator under three different operating conditions in a rural water supply system. Free and total chlorine were measured entering the storage tank, leaving the storage tank, and at three households along the transmission line of the water system in the two rural indigenous communities of Calabazal and Quebrada Mina in western Panama during April-August 2011. The Ct method for disinfection was used to compare the measured free chlorine concentration to the concentration required to inactivate common pathogens found in gravity flow water systems in Panama, such as E. coli, Salmonella typhi, Hepatitis A, Giardia lamblia, and E. histolytica, as well as other pathogens of interest to the global health community, such as Vibrio cholerae and Rotavirus. When the chlorine tablet was sealed in a plastic wrapper prior to use to prevent contact with humid surroundings, the chlorine was able to dissolve in seven days instead of three hours into the transmission line. The use of one tablet, sealed in a plastic wrapper before use, was able to obtain the required free chlorine concentration estimated to disinfect E. coli, Vibrio cholerae, Rotavirus, Salmonella typhi, and Hepatitis A. However, it did not achieve a free chlorine concentration above 0.27 mg/L needed to inactivate Giardia lamblia nor above 0.35 mg/L needed to inactivate E. histolytica. The use of three properly stored tablets in the chlorinator was able to provide a free chlorine concentration above 0.35 mg/L for only one day, reaching 0.37 mg/L, before falling below 0.35 mg/L to a level of 0.26 mg/L the next day. The study suggests that with three tablets the in-line PVC chlorinator can be an effective technology if slightly more free chlorine concentration can enter the system. The cost of this technology could be allocated to every owner with a house connection in the communities of Calabazal and Quebrada Mina by increasing their monthly tariff by $1 each month.

Access to safe drinking water has a direct effect on improving human health and their quality of life. One country still struggling with providing access to safe drinking water to all of its population is Panama. Panama's largest indigenous group, the Ngöbe people, is disproportionately affected by lack of access to safe drinking water. One way Panama's Ministry of Health (MINSA) is attempting to increase access to safe drinking water to the Ngöbe people is by disinfecting the water already captured by rural gravity fed water systems constructed within in the Ngöbe-Bugle reservation. This is accomplished using an in-line chlorinator specifically designed to accommodate locally manufactured calcium hypochlorite tablets as a source of chlorine. However, in this study it was hypothesized that the current way MINSA is implementing the in-line chlorinator was ineffective both at educating communities on knowledge of chlorination and in chlorinating water in their water distribution systems. This study investigated MINSA's implementation method and then compared it to a new method of implementation that was based on a newly developed disinfection field guide created by the author of this thesis. The motivation of this study was to improve this process of implementation which could lead to more effective chlorination thereby decreasing illness caused by waterborne pathogens. Each implementation method investigated attempted to disseminate knowledge of chlorination to community members through a seminar. The MINSA seminar was presented by a MINSA health practitioner and a newly developed seminar was presented by this thesis's author. A survey was developed to assess the knowledge of chlorination of community members after they attended a seminar. Results showed that community members who attended the new seminar on average answered 20 of the 22 questions of the administered survey more correctly than community members attending the MINSA seminar. Additionally, based on the average correct response of community members to survey questions, participants in the new seminar answered more questions correctly compared to participants in the MINSA seminar in all sections of the survey, 32% greater in the "General Knowledge" section; 43% greater in the "MINSA Specific" section; and 36% greater over the total survey. This higher score by new seminar participants suggests that the new seminar is better at educating community members on knowledge of chlorination. An assessment of each implementation method to effectively chlorinate the studied community's water distribution systems was also completed. This was done by measuring the free chlorine residual of water leaving the studied community's storage tank and entering the distribution system over one week. These concentration values were multiplied by a calculated chlorine contact time of the studied system's distribution system to determine Ct values. Measured Ct values were compared to literature guidelines that provide information on what Ct values will kill commonly found waterborne pathogens in the region. Calculated Ct values above a critical literature value of 40.0 min-mg/L Cl2 were determined to be effectively chlorinating a system's water. Results showed that when using the MINSA implementation method the required Ct level of 40.0 min-mg/L Cl2 was never met at any time during the week. However when using the new implementation method, the required Ct level of 40.0 min-mg/L Cl2 was met at all points during the week except one when tested on the last day where the Ct value was found to be 35.9 min-mg/L Cl2. These results suggest the new implementation method is more effective at chlorinating rural gravity fed water systems in the region compared to the previous implementation method.

Infertility is defined as not being able to conceive after 1 year of regular intercourse without use of contraception. Unexplained infertility is a diagnosis given to couples where the reason to infertility cannot be clarified even after the routine examination. Undefined infertility is a common and growing problem because most people are not aware of the fact that fertility decreases after the age of 35. Hyper- and hypothyroidism has been known to affect the menstrual cycle as well as increased risk of miscarriage. However, the specific effect of thyroid hormones on infertility has not yet been clarified. This study aims to compare the gene expression of two thyroid hormone receptors DIO2 and MOT8 in human endometrium and fallopian tube tissue from two phases of the menstruation cycle, follicular phase and lutheal phase. The methods used were RT-qPCR and immunohistochemistry, which showed a statistically significant difference in the expression of DIO2 and MOT8 between fallopian tube tissue and endometrium, but not between follicular and lutheal phase. However, MOT8 seemed to have a tendency to be down-regulated in the follicular phase but the results need to be validated with different endogenous controls and larger study groups.

BACKGROUND: In order to better understand the multifactorial nature of osteoporosis, animal models are utilized and compared to healthy controls. Female sheep are well established as a model for osteoporosis induced by ovariectomy, calcium and vitamin D low diet, application of steroids, or a combination of these treatments. Transcriptional studies can be performed by applying quantitative real time PCR (RT-qPCR). RT-qPCR estimates mRNA-levels of target genes in relation to reference genes. A chosen set of reference genes should not show variation under experimental conditions. Currently, no standard reference genes are accepted for all tissue types and experimental conditions. Studies examining reference genes for sheep are rare and only one study described stable reference in mandibular bone. However, this type of bone differs from trabecular bone where most osteoporotic fractures occur. The present study aimed at identifying a set of reference genes for relative quantification of transcriptional activity of ovine spine bone and ovine in vitro differentiated mesenchymal stromal cells (MSC) for reliable comparability. METHODS: Twelve candidate reference genes belonging to different functional classes were selected and their expression was measured from cultured ovMSCs (n = 18) and ovine bone samples (n = 16), respectively. RefFinder was used to rank the candidate genes. RESULTS: We identified B2M, GAPDH, RPL19 and YWHAZ as the best combination of reference genes for normalization of RT-qPCR results for transcriptional analyses of these ovine samples. CONCLUSION: This study demonstrates the importance of applying a set of reference genes for RT-qPCR analysis in sheep. Based on our data we recommend using four identified reference genes for relative quantification of gene expression studies in ovine bone or for in vitro experiments with osteogenically differentiated ovine MSCs.

BACKGROUND: In order to better understand the multifactorial nature of osteoporosis, animal models are utilized and compared to healthy controls. Female sheep are well established as a model for osteoporosis induced by ovariectomy, calcium and vitamin D low diet, application of steroids, or a combination of these treatments. Transcriptional studies can be performed by applying quantitative real time PCR (RT-qPCR). RT-qPCR estimates mRNA-levels of target genes in relation to reference genes. A chosen set of reference genes should not show variation under experimental conditions. Currently, no standard reference genes are accepted for all tissue types and experimental conditions. Studies examining reference genes for sheep are rare and only one study described stable reference in mandibular bone. However, this type of bone differs from trabecular bone where most osteoporotic fractures occur. The present study aimed at identifying a set of reference genes for relative quantification of transcriptional activity of ovine spine bone and ovine in vitro differentiated mesenchymal stromal cells (MSC) for reliable comparability. METHODS: Twelve candidate reference genes belonging to different functional classes were selected and their expression was measured from cultured ovMSCs (n = 18) and ovine bone samples (n = 16), respectively. RefFinder was used to rank the candidate genes. RESULTS: We identified B2M, GAPDH, RPL19 and YWHAZ as the best combination of reference genes for normalization of RT-qPCR results for transcriptional analyses of these ovine samples. CONCLUSION: This study demonstrates the importance of applying a set of reference genes for RT-qPCR analysis in sheep. Based on our data we recommend using four identified reference genes for relative quantification of gene expression studies in ovine bone or for in vitro experiments with osteogenically differentiated ovine MSCs.