Assessment of stool collection and storage conditions for in vitro human gut model studies.

BACKGROUND: The role of the gut microbiota in health and disease is becoming increasingly apparent. Faeces is the most accessible sample to collect from human volunteers for studying the gut microbiota. However, the impact of stool collection and storage conditions on microbial and metabolic profiles have not been fully evaluated. By understanding the effect of different stool collection and storage conditions on microbial and metabolic composition, we can consider these parameters in the design of in vitro fermentation studies. METHODS: Stool samples from 3 volunteers were stored under 5 different conditions to mimic methods that researchers may use to collect and store stool samples for study of the gut microbiota, including: fresh sample used within 10 min; stored on wet ice (4 °C) for 60 min; stored in an anaerobic chamber in a temperature-controlled bag (4 °C) for 60 min; freezing at -20 °C for 60 min and freezing at -20 °C for 60 min and then at -80 °C for 2 weeks. The stored samples were added to basal medium in batch culture fermenters alone (negative control) or with 5 g 2'-Fucosyllactose (2'FL) Human Milk Oligosaccharide (HMO) (as a positive fermentation control). Samples were collected at 3 timepoints (0, 12 and 24 h) for analysis by Flow Cytometry-Fluorescent In Situ Hybridisation (FC-FISH) and 1H-Nuclear Magnetic Resonance (NMR) spectroscopy to assess the impact on microbial and metabolic profiles, respectively. RESULTS: Freezing stool significantly impacted microbial numbers and activity during in vitro fermentations, whereas storing the stool on wet ice (4 °C) or in an anaerobic chamber at 4 °C for 60 min had minimal effects on microbial and metabolic profiles throughout the 24 h batch culture fermentation experiments. DISCUSSION: For in vitro batch culture fermentation studies where it may not be practical or possible to use fresh stool, either storing the stool on wet ice (4 °C) or in an anaerobic chamber at 4 °C for 60 min could be plausible alternatives to maintain microbial and metabolic profiles for analysis.

Development of a Pipeline for Exploratory Metabolic Profiling of Infant Urine.

Numerous metabolic profiling pipelines have been developed to characterize the composition of human biofluids and tissues, the vast majority of these being for studies in adults. To accommodate limited sample volume and to take into account the compositional differences between adult and infant biofluids, we developed and optimized sample handling and analytical procedures for studying urine from newborns. A robust pipeline for metabolic profiling using NMR spectroscopy was established, encompassing sample collection, preparation, spectroscopic measurement, and computational analysis. Longitudinal samples were collected from five infants from birth until 14 months of age. Methods of extraction and effects of freezing and sample dilution were assessed, and urinary contaminants from breakdown of polymers in a range of diapers and cotton wool balls were identified and compared, including propylene glycol, acrylic acid, and tert-butanol. Finally, assessment of urinary profiles obtained over the first few weeks of life revealed a dramatic change in composition, with concentrations of phenols, amino acids, and betaine altering systematically over the first few months of life. Therefore, neonatal samples require more stringent standardization of experimental design, sample handling, and analysis compared to that of adult samples to accommodate the variability and limited sample volume.

Urban African American women's explanations of recurrent chlamydia infections.

OBJECTIVE: To explore reasons for the high chlamydia recurrence rate among African American (AA) urban women. DESIGN: In this phenomenological qualitative study, young AA urban women with recurrent chlamydia were interviewed using open-ended questions guided by the conceptual framework of the health belief model (HBM). SETTING: The study was set in three urban health clinics in Michigan. PARTICIPANTS: Ten African American adolescents, age 15 to 19, participated. METHODS: In face-to-face recorded interviews, participants shared their personal experiences and viewpoints on what led to their recurrent chlamydia infections. The data were transcribed and analyzed through hand coding and NVivo 8 a qualitative software package. RESULTS: Overall, participants demonstrated significant knowledge deficits about the seriousness of chlamydia compared to other sexually transmitted infections (STIs). After reinfection, their perceived susceptibility changed: condom use was seen as beneficial and perceived barriers to condom use diminished as participants gained a new sense of empowerment. CONCLUSION: Chlamydia infection among African American urban adolescents is nearly 3 times that of the general population. Lack of education is still a barrier to STI prevention. Participants reported a desire to receive counseling and support from the health care staff. A STI care model that includes education, counseling, and regular screening of high-risk adolescents should be considered. Further research, using the HBM or similar theoretical models, are needed to gauge the success of any planned or implemented intervention.

The relationship of self-esteem and risky sexual behaviors in young African-American women.

In the United States, African-American women are at disproportionate risk for contracting human immunodeficiency virus (HIV) and face the most profound burden of HIV infection. Reducing the risk of exposure to HIV in African-American women is a priority for health-care providers. The findings of this study add to the existing literature by examining the relationship of self-esteem and risky sexual behaviors in young African-American women. Lack of self-esteem was one of the themes that emerged from a larger study that investigated how African-American women define HIV-risky behavior. In the current study, quantitative and qualitative data were collected from a convenience sample of 33 African-American women (N = 33) from three metropolitan regions within Michigan. Findings highlight the importance of understanding the relationship between self-esteem and its implications for HIV/sexually transmitted disease (STD) prevention.

Reducing HIV in Michigan African American young adult women.

African American (AA) women are at disproportionate risk for contracting HIV, which has reached epidemic proportions, especially in women of color. Reducing the risk of exposure to HIV in AA women is a priority for health care providers. Despite the many studies conducted on HIV in the AA community, factors that influence sexual risk taking, such as non-condom use by AA women, have yet to be understood. The purpose of this study was to investigate how AA women define HIV risky behavior. Quantitative and qualitative data were collected from a convenience sample of AA women (N = 33) from three metropolitan regions within Michigan. The results offer important insight into how AA women define HIV risky behavior as well as highlight behaviors that are amenable to intervention.

Differences in veterans' and nonveterans' end-of-life preferences: a pilot study.

BACKGROUND: Investigators conducting focus groups on end-of-life preferences noted that veterans voiced opinions that strongly differed from those of nonveterans. OBJECTIVE: The objective of this study was to further explore differences between veterans' and nonveterans' end-of-life preferences. METHODS: Ten focus groups and a pilot survey were conducted. SETTING AND SAMPLE: The focus groups consisted of Arab Muslims, Arab Christians, Hispanics, blacks, and whites stratified by gender (n = 73). Fifteen male veterans were included across all five racial groups. MEASURES: A moderator discussion guide was used to lead the focus groups and a pilot survey asked about demographic information and end-of-life preferences. RESULTS: Veterans were more likely to be married (p < 0.05) and less connected to their cultural group (p < 0.05) than nonveterans. The focus group results indicated that veterans in this study were more likely to oppose the use of heroic measures compared to nonveterans. More so than nonveterans, veterans felt that their doctors should be frank and open (p < 0.05) were strongly in favor of do-not-resuscitate (DNR) orders (p < 0.10), yet were less likely to have a proxy (p < 0.10) or durable power of attorney p < 0.01). Comparing end-of-life preferences, veterans felt less strongly than nonveterans about remembering personal accomplishments (p < 0.05), being listened to (p < 0.05), being with friends (p < 0.01), or being comfortable with their nurse (p < 0.05), but did want to be around their pets at the end of life p < 0.10). IMPLICATIONS: The Department of Veterans Affairs is in a unique position to improve endof- life care for veterans. Providing end-of-life care that is congruent with the veteran's wishes can improve satisfaction and increase cost effectiveness by eliminating unacceptable services.

Racial/ethnic preferences, sex preferences, and perceived discrimination related to end-of-life care.

This study investigated racial/ethnic preferences, sex preferences, and perceived discrimination related to end-of-life care. Ten focus groups and a follow-up survey were conducted to obtain in-depth information on end-of-life preferences across five racial/ethnic groups in Michigan stratified by sex. There were 73 focus group participants, including Arab Muslims, Arab Christians, Hispanics, blacks, and whites. The mean age+/-standard deviation was 67+/-8.5 (range 50-83). A focus group screener was used to recruit participants. A moderator discussion guide was used to guide the focus groups. A take-home questionnaire asked about demographic information and end-of-life issues. Arab Americans were in favor of making peace on earth and were against assisted suicide, extending life artificially, nursing homes, and telling the patient "bad news." Hispanic and black women were against assisted suicide and in favor of extending life, whereas the men in these groups felt the opposite. Hispanic women spoke of not wanting a feeding tube and would consider alternative medicine. Blacks were least opposed to nursing homes. For whites, it was important to have choices. When asked about discrimination related to end-of-life care, Muslim women spoke of cultural barriers, blacks spoke of inequities in the past, and whites spoke of age discrimination and abandonment when dying. As the population becomes more diverse and continues to age, it will be important to provide culturally and sex-sensitive end-of-life interventions to increase patient/family satisfaction and allocate resources appropriately.

Older African Americans' attitudes and caregiving: is there a connection?

The purpose of this non-experimental descriptive study was to explore the attitudes of older (> or =50 years old) African Americans toward and their willingness to care for people with Acquired Immune Deficiency Syndrome (AIDS). Results from this study suggest that this population has generally tolerant (empathetic) attitudes towards people with AIDS (PWA). Knowing someone with AIDS has a positive correlation with a willingness to care for someone with AIDS. Those individuals who reported a willingness to care for someone with AIDS were more likely to have more tolerant attitudes towards PWA. Recognizing the influence of older African Americans' attitudes towards PWA and their subsequent willingness to care for this population will give direction for further advanced nursing actions and research. Specifically, it will help improve family involvement as a component of the patient's support network.

Comparison of quality-of-hypertension-care indicators for groups treated by physician versus groups treated by physician-nurse team.

PURPOSE: To determine whether the type of health care provider (i.e., physician versus physician-nurse team) affected the quality of hypertension care given to two groups of randomly selected adult women. DATA SOURCES: Three indicators measured the quality of hypertension care: blood pressure control level, knowledge of hypertension, and discussion about blood pressure medications with the health care provider(s). Blood pressure readings were taken with a 24-hr ambulatory blood pressure monitor, and demographic data from survey results taken at orientation and researcher-collected data on posttreatment knowledge of hypertension and cognitive representations of hypertension were gathered. Chi-square and t tests were used to analyze the data. CONCLUSIONS: The group whose care was managed by a physician-nurse team demonstrated lower means for 24-hr systolic blood pressure and diastolic blood pressure (systolic: M = 132, SD = 14.9; diastolic: M = 75, SD = 11.3) than the group whose care was managed only by one or more physicians (systolic: M = 136, SD = 13.4; diastolic: M = 79, SD = 11.24). Also, the group whose care was managed by a physician-nurse team revealed significantly higher scores for discussion of blood pressure medication than the group whose care was managed only by one or more physicians. There were no group differences for knowledge of hypertension. IMPLICATIONS FOR PRACTICE: Nurses qualified to assist with meeting the needs of hypertension clients in primary care settings can positively affect clients' knowledge about blood pressure medication and--perhaps as a result of this knowledge--how well the clients control their blood pressure.

Biosynthesis of triacylglycerol in the filamentous fungus Mucor circinelloides.

Lipid metabolism was studied in 2-d-old liquid cultures of Mucor circinelloides grown at 25 degrees C. Under these conditions, oil accumulated to 0.5 g l-1 with a gamma-linolenic acid content (gamma 18:3) of 60 mg l-1. The major labelled lipids in cultures incubated with [14C]acetate were triacylglycerol (TAG), phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The proportion of label declined in the phospholipids and increased in TAG with time. [14C]18:1 and [14C]18:2 rapidly appeared in PC and PE and later accumulated in [14C]gamma 18:3. TAG-synthesizing capacity was greatest in the microsomal membrane fraction, which accumulated high levels of phosphatidic acid in the presence of glycerol 3-phosphate and acyl-CoA substrates at pH 7.0. Further metabolism of phosphatidic acid to diacylglycerol and TAG was achieved by increasing the pH to 8.0. Lysophosphatidic acid: acyl-CoA acyltransferase (LPAAT) activity was particularly high and may have accounted for the rapid accumulation of phosphatidic acid in the membranes. The glycerol-3-phosphate: acyl-CoA acyltransferase (GPAAT) and LPAAT were non-specific for a range of saturated and unsaturated species of acyl-CoA although the GPAAT showed a marked selectivity for palmitoyl-CoA and the LPAAT for oleoyl- and linoleoyl-CoA. gamma-Linolenic acid was detected at all three positions of sn-TAG and was particularly enriched at the sn-3 position. The preparation of active in vitro systems (microsomal membranes) capable of the complete biosynthetic pathway for TAG assembly may be valuable in understanding the assembly of oils in future transgenic applications.