Absolute quantification of eleven A, D, E and K vitamers in human plasma using automated extraction and UHPLC-Orbitrap MS.

Fat-Soluble Vitamers [FSV] deficiencies and hypervitaminosis are associated with lifestyle diseases such as cardiovascular disease, diabetes, and cancer. Quantification of FSV and their metabolites in plasma has proved to be one of the most demanding analytical chemistry challenges. Current FSV quantification methods are compromises between breadth of coverage and sensitivity across the physiological range. Here, we developed and validated a sensitive, robust, semi-automated method using liquid-liquid extraction coupled with LC-ESI-MS/MS to quantify 11 FSV across their physiological concentrations in plasma. The addition of Phree® phospholipid removal plates as the last step in the extraction process reduced matrix effects, improving precision, recoveries, and the method's final sensitivity. This method can detect and quantify: retinol, retinoic acid, retinyl palmitate, 25 hydroxyvitamin D3 [25-OH-D3], 1-α-25-dihydroxy-D3, α-tocopherol, γ-tocopherol, α-tocotrienol, phylloquinone [K1], Menatetrenone [MK-4], and menaquinone-7 [MK-7].The Instrument Quantitation Limit [IQL]s for retinol (64.1 ng/mL), 25-OH-D3 (10.2 ng/mL), and α-tocopherol (3000 ng/mL) can detect clinical deficiencies. Our automated method will assist in the understanding of the complex interaction between these compounds and their possible role in health and disease.

LC-MS/MS quantification of fat soluble vitamers - A systematic review.

Fat soluble vitamers (FSV) are several biochemically diverse micronutrients essential for healthy development, growth, metabolism, and cell regulation. We cannot synthesize FSV completely or at the required concentrations. Deficiency or excess of FSV can result in many health problems. Plasma is the most accessible sample matrix for the quantification of FSV. However, due to its complexity and other analytical challenges (e.g., FSV sensitivity to light, oxygen, heat, pH, chemical heterogeneity, standard availability), developing a method for the simultaneous quantification of multiple FSV at physiological concentrations has been challenging. In this systematic review, we examine the parameters and criteria used in existing Liquid Chromatography with tandem Mass Spectrometry (LC-MS/MS) methods for FSV quantification to the extraction method, chromatographic resolution, matrix effects, and method validation as critical to a sensitive and robust method. We conclude that the final FSV method sensitivity is predominantly based on aforementioned criteria and future method development using LC-MS/MS will benefit from the application of this systematic review.

The gathering firestorm in southern Amazonia.

Wildfires, exacerbated by extreme weather events and land use, threaten to change the Amazon from a net carbon sink to a net carbon source. Here, we develop and apply a coupled ecosystem-fire model to quantify how greenhouse gas-driven drying and warming would affect wildfires and associated CO2 emissions in the southern Brazilian Amazon. Regional climate projections suggest that Amazon fire regimes will intensify under both low- and high-emission scenarios. Our results indicate that projected climatic changes will double the area burned by wildfires, affecting up to 16% of the region's forests by 2050. Although these fires could emit as much as 17.0 Pg of CO2 equivalent to the atmosphere, avoiding new deforestation could cut total net fire emissions in half and help prevent fires from escaping into protected areas and indigenous lands. Aggressive efforts to eliminate ignition sources and suppress wildfires will be critical to conserve southern Amazon forests.

Differential diagnosis between a granuloma and radicular cyst: effectiveness of magnetic resonance imaging.

AIM: To investigate the diagnostic reliability and accuracy of magnetic resonance imaging (MRI) to differentiate periapical lesions of endodontic origin and to compare the results with histopathological information. METHODOLOGY: The radiolucent periapical jaw lesions of 34 patients, which were surgically enucleated, were investigated by two radiologists using MRI, based on the same six criteria, to categorize the lesions as granulomas, radicular cysts or others. After apicoectomies, two oral pathologists (blinded to the radiologist's diagnoses) analysed all specimens by referring to seven specific parameters and diagnosed the specimens as granulomas, radicular cysts or other conditions. The inter-rater agreements between the radiologists and pathologists in terms of MRI and histological diagnoses, respectively, along with the discriminant power of the adopted criteria and the accuracy of the MRI assessments compared with the histopathological results, were calculated. Cohen's kappa test was adopted to examine inter-rater agreement between the two radiologists and two pathologists. Guttman's lambda coefficient (λ6 ) was used to evaluate the internal consistency of the items used for the differential diagnosis by radiologists. The accuracy resulted from a receiver operator characteristic (ROC) analysis. RESULTS: A strong inter-rater reliability was observed between the two radiologists (k-statistic = 0.86, P = 0.0001) and the two pathologists (k-statistic = 0.88, P = 0.0001). The internal consistency of the diagnostic items was 0.605 for cysts and 0.771 for granulomas. The accuracy (true positives plus true negatives) of the radiologists was greater than that of the pathologists based on analysis (area under the curve = 0.87 and 0.91, respectively). CONCLUSIONS: The reliability and accuracy of MRI were high and comparable to histopathological reliability, highlighting the usefulness of this noninvasive technique as a pre-treatment diagnostic method for periapical endodontic lesions.

Comparison of transvaginal sonography and double-contrast barium enema for diagnosing deep infiltrating endometriosis of the posterior compartment.

OBJECTIVES: To compare the diagnostic accuracy of transvaginal sonography (TVS) and double-contrast barium enema (DCBE) in the preoperative detection of deep infiltrating endometriosis (DIE) of the posterior compartment. METHODS: This was a prospective study of 69 consecutive patients with results of pelvic examination or symptoms suggestive of DIE of the posterior compartment. TVS and DCBE were performed before surgery by two groups of physicians specialized in endometriosis, each blinded to the results of the other technique. Imaging data were compared with histopathologic analysis of the resected specimen (gold standard). Sensitivity, specificity, positive and negative predictive values and test accuracies were calculated for both imaging modalities. RESULTS: Sixty seven of the 69 women had a nodule of DIE confirmed at laparoscopy and histopathologic examination. TVS diagnosed DIE in 57 (85%) of these patients, while DCBE revealed the presence of the lesion in 24 (36%) women. For the diagnosis of posterior DIE, TVS and DCBE had, respectively, a sensitivity of 85% and 36%, specificity of 100% and 100%, positive predictive value of 100% and 100%, negative predictive value of 17% and 4% and accuracy of 85.5% and 38%. In patients with pure bowel DIE the sensitivity was 91% and 43%, specificity was 100% and 100%, positive predictive value was 100% and 100%, negative predictive value was 29% and 6% and accuracy was 91% and 45%, respectively. CONCLUSIONS: TVS has a much higher sensitivity than does DCBE in detecting the presence of posterior DIE and should thus be regarded as the imaging modality of choice when there is clinical suspicion of the disease.

Using focus groups in community-based participatory research: challenges and resolutions.

A community-based participatory approach requires that community members be involved in all phases of the research process. We describe three focus group studies with American Indians in Kansas and Missouri, using a newly developed method of conducting and analyzing focus groups with community input (72 focus groups, 519 participants). We conducted two needs assessment studies focused on barriers to breast and colorectal cancer screening and one study focused on Internet use for gathering health information. Community members and researchers collaborated to develop guides for the focus group moderators. Community organizations and our community advisory board conducted recruitment, and we trained and employed community members as moderators, assistant moderators, and analysts. Our community partners also helped with dissemination of research findings to their constituents. The methodologic approach and data from these three studies will allow us to more appropriately address health disparities in the American Indian community, with full community support for our research.

The Culture of Health Survey: a qualitative assessment of a diabetes prevention coalition.

In the past two decades, the fields of public health and social services have increasingly turned toward collaborative and community-based approaches to address complex health and social issues. One aspect of these approaches has been the development and implementation of community coalitions. Coalitions have been used to successfully address a wide range of issues, including cancer prevention, tobacco use, HIV/AIDS, youth violence, heart disease, diabetes, and sexual exploitation of youth runaways. In south Tucson, Arizona the SEAH coalition was developed to address diabetes and substance abuse prevention. Using a qualitative interview guide, the Culture of Health Survey, this study was aimed at identifying community perceptions of the coalition and its effectiveness in the areas of community leadership, partnerships, trust, and movement towards positive change. We also sought to document the dissemination, throughout a community, of information on the activities and functioning of a community based coalition and whether or not it was seen as one that held fast to the community values and not to individual agendas. Results highlight the importance of outreach, education, trust, and partnerships in promoting diabetes prevention through a community coalition.

Male preventive health behaviors: perceptions from men, women, and clinical staff along the U.S. Mexico border.

Mexican American males have higher levels of total cholesterol and triglycerides, higher body mass indexes, and a higher prevalence of diabetes than do non-Hispanic White males. They are the least likely Hispanic subgroup to be insured, to have recently visited a physician, or to have preventive exams. To explore factors related to the use of preventive exams among mature men, and specifically among Mexican American men residing along the Arizona, United States/Sonora, Mexico border, information on barriers and motivating factors to male participation in preventive screening exams was collected. Interviews were conducted with mature men and women from a single border community and with clinical staff from three different border communities who deliver services to similar populations. Responses were triangulated. Common themes identified include health education/information/advertisement and female/family support as motivating factors and machismo/denial/fatalism as a barrier to male health-seeking behavior.

Psychometric evaluation of the marijuana and stimulant effect expectancy questionnaires for adolescents.

Alcohol expectancies are important in the mediation and prediction of alcohol use. Expectancies for the effects of other drugs, although less well delineated, appear equally important. Therefore, development and validation of expectancy measures for drugs other than alcohol is necessary for evaluating the importance of these constructs. We examined the factor structure, reliability, and validity of the Marijuana Effect Expectancy Questionnaire (MEEQ) and the Stimulant Effect Expectancy Questionnaire (SEEQ) in clinical and community samples of adolescents as they moved into young adulthood (N=279). Confirmatory factor analyses (CFAs) supported the a priori factors, and we found good reliability for most individual scales. Temporal stability and convergent and discriminant validity of drug effect expectancies were supported in this sample of adolescents and young adults. Drug effect expectancies were associated with drug preference and drug use patterns over 2 years. Use of these measures may aid our understanding of the etiology and course of marijuana and stimulant involvement during adolescence and young adulthood.

Wernicke's encephalopathy in a child: case report and MR findings.

We report a child affected by Wernicke's encephalopathy (WE), which was unsuspected clinically. MRI suggested the correct diagnosis and prompted appropriate thiamine replacement. WE is a difficult condition to recognise, especially in children, and MRI may be useful in the diagnosis of the disease.

Kawasaki syndrome in an adult: case report and review of the literature in adolescents and adults.

Kawasaki syndrome in adults is very rare, with fewer than 50 cases reported in the English-language literature. We describe the case of a physician with Kawasaki syndrome and summarize the clinical features and treatment of 11 patients in the literature since the last review in 1994. Our patient presented with high fever, conjunctivitis, and arthralgias, then developed progressive toxicity with oral lesions, cervical adenopathy, and desquamation of the fingers and toes. No exanthematous rash or coronary artery aneurysms were found. Recovery was rapid after therapy with aspirin and intravenous immunoglobulin (IVIg). The diagnosis of Kawasaki syndrome depends on clinical criteria and the exclusion of other diseases. This diagnosis can be challenging to make in an adult, particularly when it presents without all typical features. Kawasaki syndrome must be considered nonetheless in an adult with unexplained fever of more than 5 days duration, because early diagnosis and combination therapy with aspirin and IVIg can prevent the life-threatening complication of coronary artery aneurysms.

Cluster of serogroup C meningococcal disease associated with attendance at a party.

BACKGROUND: An unexplained increase has occurred in the incidence of invasive meningococcal disease in adolescents and young adults. METHODS: We investigated a cluster of serogroup C meningococcal disease in 3 previously healthy young adults who had attended a party in Maryland. Molecular subtyping was done on the isolates from the 3 cluster cases and 4 control isolates by pulsed-field gel electrophoresis (PFGE). The only common exposure was attendance at the party, where a large number of people reportedly smoked tobacco or marijuana and/or drank alcohol. RESULTS: The PFGE analysis of the 3 case isolates showed identical molecular subtypes. CONCLUSION: This investigation strongly suggests that transmission of the cluster strain occurred at the party. Transmission may have occurred in part as a result of the recently described risk factors of binge drinking and smoking. Taken together, these findings suggest that some of the recent increase in invasive meningococcal disease may be due to modifiable risk factors.

Effect of virginiamycin on ruminal fermentation in cattle during adaptation to a high concentrate diet and during an induced acidosis.

The objective of Exp. 1 was to compare the effects of virginiamycin (VM; 0, 175, or 250 mg x animal(-1) x d(-1)) and monensin/tylosin (MT; 250/ 90 mg x animal(-1) x d(-1)) on ruminal fermentation products and microbial populations in cattle during adaptation to an all-concentrate diet. Four ruminally cannulated, Holstein steers were used in a 4x4 Williams square design with 21-d periods. Steers were stepped up to an all-concentrate diet fed at 2.5% of BW once daily. Ruminal pH, protozoal counts, and NH3-N and VFA concentrations generally were unaffected by VM or MT. Mean counts of Lactobacillus and Streptococcus bovis were lower (P<.05) for VM-treated compared with control or MT-treated steers. Both VM and MT prevented the increase in Fusobacterium necrophorum counts associated with increasing intake of the high-concentrate diet observed in the control. The objective of Exp. 2 was to compare the effects of VM and MT on ruminal pH, L(+) lactate and VFA concentrations, and F. necrophorum numbers during carbohydrate overload. Six ruminally cannulated Holstein steers were assigned randomly to either the control, VM (175 mg/d), or MT (250 + 90 mg/d) treatments. Acidosis was induced with intraruminal administration of a slurry of ground corn and corn starch. The VM and MT premixes were added directly to the slurry before administration. Carbohydrate challenge induced acute ruminal acidosis (pH was 4.36 and L (+) lactate was 19.4 mM) in controls by 36 h. Compared with the controls, steers receiving VM or MT had higher (P<.05) ruminal pH, and the VM group had a lower (P<.05) L (+) lactate concentration. Fusobacterium necrophorum numbers initially increased in VM- and MT-administered steers. In the control steers, F. necrophorum was undetectable by 36 h. Virginiamycin seemed to control the growth of ruminal lactic acid-producing bacteria and, therefore, has the potential to moderate ruminal fermentation in situations that could lead to rapid production of lactic acid.

Adolescent alcohol and drug abuse and health.

PURPOSE: To examine the relationship of adolescent alcohol and drug use over a 5-year period to cumulative health problems in late adolescence and young adulthood. METHODS: We prospectively examined self-reported health problems in a sample of adolescents, some of whom received treatment for substance use disorders and had consistently poor substance use outcomes (n = 38), some of whom received treatment for substance use disorders and had positive substance use outcomes (n = 30), and a low alcohol and drug use community comparison group (n = 48). Data regarding health-related problems of these adolescents (mean, 15.9 years; 83% Caucasian; 56.5% female) were collected at 2, 4, and 6 years following initial assessments. RESULTS: Alcohol and/or drug involvement severe enough to warrant treatment during adolescence was associated with more cumulative health problems and severe health problems for girls and more cumulative health problems for boys. Protracted and continuous abuse of alcohol and drugs was associated with more cumulative and severe health problems for girls and more severe health problems for boys. CONCLUSIONS: These results suggest that significant health problems and concerns are related to both brief and protracted alcohol and drug abuse during adolescence. Health problems will likely become even more evident as early-onset, chronic substance abusers continue to age.

Male caregivers' use of formal support.

The purpose of this study was to explore male caregivers' perceptions of formal support. The men were caregivers of adults with cognitive impairment. The study involved secondary analysis of interview data on perceptions of support that had not been previously analyzed from the perspective of formal support. Techniques of data analysis used in grounded theory were applied to 62 guided interactive interviews conducted with 24 male caregivers during a period of 18 months. The men experienced a process of making concessions for care in the following four sequential stages: resisting, giving in, opening the door, and making the match. Personal barriers deterred the caregivers from seeking help, and once the caregivers engaged formal help the influence of characteristics of the health care system and individual staff members had both enabling and disabling effects. For caregivers whose use of formal support involved admitting their relative to a long-term care facility, the phase of making the match was followed by redesigning their caregiver role.

Giant Cell Arteritis in AJ Americans.

Giant cell arteritis is often considered less common in African Americans. In 1985, we reviewed 34 African American patients with biopsy-proven temporal arteritis. We report on 9 additional biopsy positive patients seen at the Washington Hospital Center (WHO between 1988 and 1996.Clinical presentations were similar to those reported in previous series, with the exception that only one patient had polymyalgia rheumatica. Two patients had visual symptoms, but only one patient had permanent monocular visual loss. In combining our 9 new patients with 41 biopsy-proven, well characterized, published patients and comparing them with a representative Caucasian group, we found statistically significant differences. Male gender, anemia, and visual loss were overall more common in African Americans, whereas constitutional symptoms and polymyalgia rheumatica were less common. The detection rate of temporal artery biopsies at WHC was lower than rates reported in other studies, but they did not differ by race.We conclude that giant cell arteritis should be pursued as a diagnosis in all patients presenting with appropriate clinical features, regardless of race, and that earlier recognition may account for the lower incidence of permanent visual loss.

Hypertonic activation and recovery of system A amino acid transport in renal MDCK cells.

Amino compounds are abundant within the renal inner medulla, but their possible role during hypertonic stress is not clear. Renal epithelial Madin-Darby canine kidney cells were used to examine the osmoregulation of system A transport, a major Na(+)-dependent process for neutral amino acid transport. System A activity was markedly increased after 6 h of hypertonic challenge, and intracellular alanine content increased more than twofold. The activation of system A was reversed after 24 h of hypertonic challenge. This downregulation was accompanied by the activation of betaine transport, as measured by gamma-aminobutyric acid uptake. Extracellular betaine prevented the early activation of system A. The hypertonic activation of system A was blocked by actinomycin D and cycloheximide. When cells were returned to isotonic medium after hypertonic activation, the recovery of system A transport also was partially inhibited by actinomycin D and puromycin. The results are consistent with the possibility that hypertonicity, by disrupting a repressor protein, leads to increased synthesis of a system A-related protein. The isotonic recovery may require synthesis of new repressor proteins.