Engineering a Yellow Thermostable Fluorescent Protein by Rational Design.

Thermal green protein (TGP) is an extremely stable, highly soluble synthetic green fluorescent protein. The quantum yield of TGP is lower than the closest related natural fluorescent protein, monomeric Azami-Green. We improved the thermal recovery of TGP through the introduction of a chromophore mutation, Q66E. Furthermore, we developed a yellow thermal protein (YTP) via mutation of histidine 193 to tyrosine. Incorporation of Q66E into YTP (YTP-E) improved chemostability and pH stability. Both YTP and YTP-E have superior thermostability compared to TGP or TGP-E. These proteins offer a new option for green or yellow fluorescence under harsh chemical or thermal conditions.

Contribution of Mitophagy to Cell-Mediated Mineralization: Revisiting a 50-Year-Old Conundrum.

Biomineralization in vertebrates is initiated via amorphous calcium phosphate (ACP) precursors. These precursors infiltrate the extracellular collagen matrix where they undergo phase transformation into intrafibrillar carbonated apatite. Although it is well established that ACP precursors are released from intracellular vesicles through exocytosis, an unsolved enigma in this cell-mediated mineralization process is how ACP precursors, initially produced in the mitochondria, are translocated to the intracellular vesicles. The present study proposes that mitophagy provides the mechanism for transfer of ACP precursors from the dysfunctioned mitochondria to autophagosomes, which, upon fusion with lysosomes, become autolysosomes where the mitochondrial ACP precursors coalesce to form larger intravesicular granules, prior to their release into the extracellular matrix. Apart from endowing the mitochondria with the function of ACP delivery through mitophagy, the present results indicate that mitophagy, triggered upon intramitochondrial ACP accumulation in osteogenic lineage-committed mesenchymal stem cells, participates in the biomineralization process through the BMP/Smad signaling pathway.

Health outcomes and retention in care following release from prison for patients of an urban post-incarceration transitions clinic.

Chronic health conditions are overrepresented among prisoners who often face barriers to medical care following release. Transitions clinics seek to provide timely access to medical care following release. This retrospective cohort study investigated care delivery and health outcomes for recently released prisoners receiving care at the Bronx Transitions Clinic. Among 135 recently released prisoners, median time from release to initial medical visit was 10 days (IQ Range: 5-31). Six-month retention in care was high for HIV-infection (86%), but lower for opioid dependence (33%), hypertension (45%) and diabetes (43%). At six months, 54% of HIV-patients had a suppressed viral load, but fewer buprenorphine-treated patients reduced opioid use (19%), and fewer hypertensive and diabetic patients reached respective blood pressure (35%) and hemoglobin A1c (14%) goals. Access to medical care is necessary but not sufficient to control chronic health conditions. Additional interventions are necessary for formerly incarcerated people to achieve optimal health outcomes.

A description of an urban transitions clinic serving formerly incarcerated people.

Chronic health conditions are overrepresented among jail or prison inmates but often go untreated during incarceration and following release. We describe the Bronx Transitions Clinic, a partnership between a community-based organization and an academic medical center, which facilitates connections to medical care for formerly incarcerated people.

An in vitro study of the effect of different restorative materials on the reliability of a veneering porcelain.

STATEMENT OF PROBLEM: Implant-supported, porcelain veneered restorations experience a greater rate of porcelain fracture than tooth-supported restorations. For completely edentulous patients, one approach to minimizing porcelain fracture is to use acrylic resin in the mandible, although its efficacy is unknown. PURPOSE: The purpose of this study was to evaluate the reliability of a veneering porcelain fatigued with different restorative materials in vitro. MATERIAL AND METHODS: Fifty-nine veneering porcelain disk specimens were fabricated by layering veneering porcelain on nickel-chromium base metal alloy disks. Four groups of different indenter materials fatigued the porcelain specimens: group WC, tungsten carbide served as a control; group FC, pressed leucite glass ceramic; group NHC, nanohybrid composite resin denture tooth; and group AR, unfilled acrylic resin denture tooth. Porcelain specimens were randomly divided into 4 groups (n=14). A step-stress accelerated life-testing model was used. Use-level probability Weibull plots were generated, and the reliability of each group was estimated for a theoretical completion of 50 000 cycles at 150 N. RESULTS: Nanohybrid composite resin and unfilled acrylic resin denture tooth groups had higher reliability than tungsten carbide and leucite glass ceramic groups. No significant differences existed between the reliability of the tungsten carbide and leucite glass ceramic groups and the nanohybrid composite resin and acrylic resin denture tooth groups. CONCLUSIONS: Veneering porcelain disk specimens fatigued with the unfilled acrylic resin and nanohybrid composite resin denture tooth indenters exhibited higher reliability than the specimens fatigued with either the tungsten carbide or leucite glass ceramic indenters. All of the veneering porcelain disk specimens failed with the same mode of fracture, although the surface posttest exhibited different fracture characteristics among specimens fatigued with the 4 different materials.

Does HIV VCT reduce risk behaviors? An observational study in Guatemala City.

OBJECTIVE: We examined the impact of HIV voluntary counseling and testing on self-reported behavioral risks three months after HIV testing. DESIGN: Cohort study comparing self-reported risk behaviors prior to and three months after HIV testing. SETTING: Clinica Familiar Luis Angel Garcia, an HIV specialty clinic located in a Guatemalan National Hospital. SUBJECTS, PARTICIPANTS: 144 people undergoing HIV testing were enrolled. 44 were HIV positive. 41 HIV positive and 49 HIV negative subjects returned for follow-up interviews. INTERVENTION: All subjects were tested and received voluntary counseling regarding HIV infection, transmission, prevention, and interpretation of HIV test results. MAIN OUTCOME MEASURE: The primary study outcome measure was change in self-reported risk behaviors three months after voluntary counseling and testing. RESULTS: Men were more likely than women to report a history of sexually transmitted diseases, more than 2 sexual partners, using alcohol with sex, and receiving money for sex; they were less likely to have a regular partner. 26% of men reported non-heterosexual orientation; no woman did. Alcohol was the primary drug of abuse in both men and women. At three month follow-up HIV positive subjects showed decreases in the average number of sexual partners, use of alcohol with sex, and episodes of unprotected sex. CONCLUSIONS: Voluntary counseling and testing resulted in changes in some self-reported risk behaviors, but only among HIV positive subjects. On nearly all measures men report riskier behavior than women. Alcohol is the most commonly used drug in this population and is often used with sex.

Experience of a pediatric HIV clinic in Guatemala City.

OBJECTIVES: To describe the clinical experience of a Guatemalan pediatric HIV clinic and referral center, and fill the gap in literature available on pediatric HIV in Guatemala, a country facing a growing HIV epidemic. METHODS: Analyses were performed on data available from the clinical databases maintained by the Clínica Familiar Luis Angel García within the Hospital General San Juan de Dios in Guatemala City, Guatemala. RESULTS: From January 1997-June 2006, a total of 536 children (individuals under 13 years of age) were registered at the clinic, 54% of them female. At the initial visit, 241 were known to be HIV infected, while 295 were known to have been exposed to HIV, but were of undetermined infection status. Of the 295 with undetermined status, serostatus was determined in 173, and 57 (33%) were HIV positive. The patients came from all 24 departments of Guatemala, but the majority (64%) was from Guatemala City. Most had perinatal exposure; three patients had been sexually exposed to HIV (all male); and the mode of infection could not be determined for six children. In the cohort of children whose infection status was initially undetermined, the provision of antiretroviral (ARV) medication (both pre- and neonatal), in addition to Cesarean section, was associated with an odds ratio of 0.06 for HIV infection (P < 0.001) when compared to children who had no interventions. Highly active antiretroviral therapy (HAART) was administered to 167 HIV-infected children. There were 44 known deaths in this cohort; no deaths occurred among the children who were not infected. CONCLUSIONS: Pediatric HIV/AIDS is present in all parts of Guatemala. Programs to prevent mother to child transmission and to provide appropriate treatment to families living with HIV/AIDS must be a public health priority.

Experience of a pediatric HIV clinic in Guatemala City / Experiencia de una clínica pediátrica para el VIH en la Ciudad de Guatemala

OBJECTIVES: To describe the clinical experience of a Guatemalan pediatric HIV clinic and referral center, and fill the gap in literature available on pediatric HIV in Guatemala, a country facing a growing HIV epidemic. METHODS: Analyses were performed on data available from the clinical databases maintained by the Clínica Familiar Luis ángel García within the Hospital General San Juan de Dios in Guatemala City, Guatemala. RESULTS: From January 1997-June 2006, a total of 536 children (individuals under 13 years of age) were registered at the clinic, 54 percent of them female. At the initial visit, 241 were known to be HIV infected, while 295 were known to have been exposed to HIV, but were of undetermined infection status. Of the 295 with undetermined status, serostatus was determined in 173, and 57 (33 percent) were HIV positive. The patients came from all 24 departments of Guate mala, but the majority (64 percent) was from Guatemala City. Most had perinatal exposure; three patients had been sexually exposed to HIV (all male); and the mode of infection could not be determined for six children. In the cohort of children whose infection status was initially undetermined, the provision of antiretroviral (ARV) medication (both pre- and neonatal), in addition to Cesarean section, was associated with an odds ratio of 0.06 for HIV infection (P < 0.001) when compared to children who had no interventions. Highly active antiretroviral therapy (HAART) was administered to 167 HIV-infected children. There were 44 known deaths in this cohort; no deaths occurred among the children who were not infected. CONCLUSIONS: Pediatric HIV/AIDS is present in all parts of Guatemala. Programs to prevent mother to child transmission and to provide appropriate treatment to families living with HIV/AIDS must be a public health priority.

OBJETIVOS: Describir la experiencia de una clínica y centro de referencia pediátrico para el VIH en Guatemala y llenar el vacío existente en la literatura científica sobre la infección pediátrica por el VIH en ese país, donde se enfrenta una creciente epidemia por el VIH. MÉTODOS: SE analizaron los datos disponibles en la base de datos clínicos de la Clínica Familiar Luis Ángel García del Hospital General San Juan de Dios, Ciudad de Guatemala, Guatemala. RESULTADOS: Entre enero de 1997 y junio de 2006 en la clínica se registraron 536 menores de 13 años; 54 por ciento de los cuales eran niñas. En la visita inicial se sabía que 241 estaban infectados por el VIH y que 295 habían estado expuestos a ese virus aunque su diagnóstico era indeterminado. De estos, se determinó el estatus serológico de 173, de los cuales 57 (33 por ciento) resultaron positivos al VIH. Los pacientes procedían de los 24 departamentos de Guatemala, pero la mayoría (64 por ciento) era de la Ciudad de Guatemala. La mayoría de los pacientes había estado expuestos al VIH por la vía perinatal; tres pacientes (todos varones), por la vía sexual y no se pudo determinar la vía de infección de seis niños. En la cohorte de niños con diagnóstico inicial indeterminado, el tratamiento antirretroviral (tanto pre como posnatal) junto con el nacimiento por cesárea se asociaron con una razón de posibilidades (odds ratio) de tener la infección por el VIH de 0,06 (P < 0,001) en comparación con los niños sin estas intervenciones. El tratamiento antirretroviral de gran actividad (HAART) se administró a 167 niños infectados por el VIH. Se registraron 44 muertes en esta cohorte, ninguna en los niños sin infección. CONCLUSIONES: La infección pediátrica por VIH/sida está presente en todos los departamentos de Guatemala. Los programas para evitar la transmisión de madre a hijo y brindar el tratamiento adecuado a las familias que viven con VIH/sida son una prioridad de salud pública.

Vaginal douching among Latinas: practices and meaning.

OBJECTIVES: Vaginal douching is widely practiced by American women, particularly among minority groups, and is associated with increased risk of pelvic and vaginal infections. This research sought to investigate vaginal hygiene practices and meaning associated with them among Latina women and adolescents. Study results would guide development of an intervention to decrease douching among Latinas. METHODS: In depth qualitative interviews conducted with English- and Spanish-speaking women aged 16-40, seeking care for any reason who reported douching within the last year (n = 34). Interviews were audiotaped, transcribed and analyzed using qualitative methods. One-third of interviews were conducted in Spanish. RESULTS: Two explanatory models for douching motives emerged: one stressed cosmetic benefits; the other, infection prevention and control. Most women reported douching to eliminate menstrual residue; a small number reported douching in context of sexual intercourse or vaginal symptoms. Many were unaware of associated health risks. Respondents typically learned about douching from female family members and friends. Male partners were described as having little to no involvement in the decision to douche. Women varied in their willingness to stop douching. Two-thirds reported receiving harm reduction messages about "overdouching". About half indicated previous discussion about douching with health care providers; some had reduced frequency in response to counseling. A number of previously unreported vaginal hygiene practices and products were described, including use of a range of traditional hygiene practices, and products imported from outside the US. CONCLUSIONS: Respondents expressed a range of commitment to douching. Counseling messages acknowledging benefits women perceive as well as health risks should be developed and delivered tailored to individual beliefs. Further research is needed to assess prevalence and safety of previously unreported practices.

Using Google Earth as an innovative tool for community mapping.

Maps are used to track diseases and illustrate the social context of health problems. However, commercial mapping software requires special training. This article illustrates how nonspecialists used Google Earth, a free program, to create community maps. The Bronx, New York, is characterized by high levels of obesity and diabetes. Residents and medical students measured the variety and quality of food and exercise sources around a residency training clinic and a student-run free clinic, using Google Earth to create maps with minimal assistance. Locations were identified using street addresses or simply by pointing to them on a map. Maps can be shared via e-mail, viewed online with Google Earth or Google Maps, and the data can be incorporated into other mapping software.

An investigation of douching practices in the botánicas of the Bronx.

Douching is a common practice in women and has been associated with adverse health outcomes. In order to explore douching products and practices we conducted qualitative interviews in ten botánicas (stores that provide healing and spiritual services to immigrant communities) located in New York City. We interviewed 15 people, 14 of whom were botánica owners and employees and ten of whom were women. We found that douching was not easily separated from the more holistic concerns of botánica customers involving health, well-being and spirituality. These issues included abortion, infertility, menopause, the prevention and treatment of infections, sexuality, cleanliness, hygiene and relationship issues. The vagina was seen as a sensitive, even vulnerable part of the body, not clearly distinguished from other female organs. A variety of products were used in the vagina in the form of creams, douches, suppositories, baths and herbal steaming of the urogenital area. Alum, an astringent, was used for the purposes of vaginal tightening to enhance sexual pleasure for the partner, to make the vagina 'younger', or to hide evidence of infidelity. Botánicas are part of a complex healing system with conceptual models different from those of allopathic medicine. These models may not be unique to the botánicas.

How do clinicians manage vaginal complaints? An Internet survey.

INTRODUCTION: Although vaginal symptoms are among the most common presentations in primary care, little is known about how clinicians manage them. QUESTION: We undertook an Internet survey to determine how clinicians decided what was normal, how they evaluated vaginal complaints, and what happened when a diagnosis could not be reached. METHODS: A 27-question anonymous survey was conducted using the Medscape Internet publication platform and Medscape General Medicine , an online, peer-reviewed, open-access general medical journal. The study was targeted at physicians and nurse practitioners. RESULTS: There were 556 nurse practitioner and 608 physician respondents. Clinicians generally agreed that a midcycle increase in discharge and discharge seen at the introitus were normal; itching/irritation, odor, and a green discharge were not. There was less agreement concerning the value of a yellow discharge and "self-diagnosis" of abnormality. Eighty-two percent of MDs and 77% of NPs reported examining patients prior to treatment always or often. Only about 30% performed pH testing always or often. Ninety percent of NPs and 92% of MDs reported not making a diagnosis rarely or sometimes. When offered 4 different management strategies for patients in whom no diagnosis was made (watch and wait, symptomatic therapy, empiric antibiotics, referral), there was no clear preference. There was a weak correlation between performance of a wet mount and reports of less non-diagnosis. There was no correlation between pH and whiff testing and reports of less non-diagnosis. CONCLUSIONS: Clinicians do not always agree about what constitutes a vaginal symptom. Although most providers usually examine patients with vaginal complaints, pH and whiff testing are performed less frequently. Our data suggest the latter 2 maneuvers are not diagnostically helpful. There is no consensus regarding what to do when no diagnosis is made. Our findings suggest that the traditional approach to the evaluation of vaginal symptoms may have limited relevance for clinicians.

Evaluation of vaginal complaints.

CONTEXT: Vaginal symptoms are one of the most common reasons for gynecological consultation. Clinicians have traditionally diagnosed vaginal candidiasis, bacterial vaginosis, and vaginal trichomoniasis using some combination of physical examination, pH, the wet mount, and the whiff test. OBJECTIVES: To evaluate the role of the clinical examination and determine the positive and negative likelihood ratios (LRs) for the diagnosis of vaginal candidiasis, bacterial vaginosis, and vaginal trichomoniasis. DATA SOURCES: Using a structured literature review, we abstracted information on sensitivity and specificity for symptoms, signs, and office laboratory procedures. We chose published (1966 to April 2003) articles that appeared in the MEDLINE database and were indexed under the combined search terms of diagnosis with vaginitis, vaginal discharge, candidiasis, bacterial vaginosis, and trichomoniasis. STUDY SELECTION: Included studies of symptomatic premenopausal women seen in primary care settings. Tests were evaluated only if they would provide diagnostic information during the office visit and were compared with an acceptable criterion standard. DATA EXTRACTION: All 3 authors extracted the data and computed sensitivity and specificity from each article independently. The absence of standard definitions for symptoms and signs made it impossible to combine results across studies. DATA SYNTHESIS: Symptoms alone do not allow clinicians to distinguish confidently between the causes of vaginitis. However, a patient's lack of itching makes candidiasis less likely (range of LRs, 0.18 [95% confidence interval [CI], 0.05-0.70] to 0.79 [95% CI, 0.72-0.87]) and lack of perceived odor makes bacterial vaginosis unlikely (LR, 0.07 [95% CI, 0.01-0.51]). Similarly, physical examination signs are limited in their diagnostic power. The presence of inflammatory signs is associated with candidiasis (range of LRs, 2.1 [95% CI, 1.5-2.8] to 8.4 [95% CI, 2.3-31]). Presence of a "high cheese" odor on examination is predictive of bacterial vaginosis (LR, 3.2 [95% CI, 2.1-4.7]) while lack of odor is associated with candidiasis (LR, 2.9 [95% CI, 2.4-5.0]). Office laboratory tests, particularly microscopy of vaginal discharge, are the most useful way of diagnosing these 3 conditions. CONCLUSIONS: The cause of vaginal complaints may be easily diagnosed when typical findings appear in microscopy. However, the poor performance of individual symptoms, signs, and office laboratory tests often makes it problematic to identify the cause of vaginal symptoms.