Current advances in microbiome sciences within the US Department of Defense: part 2 - enabling technologies and environmental microbiomes.

Microbiomes involve complex microbial communities wherein the micro-organisms interact with one another as well as their associated hosts or environmental niches. Much of the characterisation of these communities and the associations have been achieved through 'omics' technologies, such as metagenomics, metaproteomics and metametabolomics, and model systems. Recent research in host-associated microbiomes has been aimed at understanding the role microbes may play in host fitness or conversely how host activities/conditions may perturb the microbial community, which can further affect host health. These studies have led to the investigation of detection, intervention or modulation methods, which may serve to provide benefits to the host and advance our understanding of microbiome associations. With the clear implications on human health and disease, the US Department of Defense (DoD) has made microbiome research a priority, with the founding of the Tri-Service Microbiome Consortium (TSMC) to enhance collaboration, coordination,and communication of microbiome research among DoD organisations and partners in academia and industry. DoD microbiome research focuses mainly on the following themes: (1) human health and performance, (2) environmental microbiomes and (3) enabling technologies. This review provides an update of current DoD microbiome research efforts centred on enabling technologies and environmental microbiomes and highlights innovative research being done in academia and industry that can be leveraged by the DoD. These topics were also communicated and further discussed in the Fifth Annual TSMC Symposium. This paper forms part of the special issue of BMJ Military Health dedicated to personalised digital technology for mental health in the Armed Forces.

Current advances in microbiome sciences within the US Department of Defense-part 1: microbiomes for human health and performance.

Microbiomes involve complex microbial communities where the microorganisms interact with one another as well as their associated hosts or environmental niches. The characterisation of these communities and associations have largely been achieved through 'omics' technologies, such as metagenomics, metaproteomics and metametabolomics, and model systems. Recent research in host-associated microbiomes have been aimed at understanding the roles microbes may play in host fitness or conversely how host activities/conditions may perturb the microbial community, which can further affect host health. These studies have led to the investigation of detection, intervention or modulation methods, which may serve to provide benefits to the host and advance our understanding of microbiome associations. With the clear implications on human health and disease, the US Department of Defense (DoD) has made microbiome research a priority, with the founding of the Tri-Service Microbiome Consortium (TSMC) to enhance collaboration, coordination and communication of microbiome research among DoD organisations and partners in academia and industry. DoD microbiome research focuses mainly on the following themes: (1) Human health and performance; (2) Environmental microbiomes; and (3) Enabling technologies. This review provides an update of current DoD microbiome research efforts centred on human health and performance and highlights innovative research being done in academia and industry that can be leveraged by the DoD. These topics were also communicated and further discussed during the fifth Annual TSMC Symposium. This paper forms part of the special issue of BMJ Military Health dedicated to Personalised Digital Technology for Mental Health in the Armed Forces.

Evaluation of the in vitro anti-hyperglycemic effect of Cinnamomum cassia derived phenolic phytochemicals, via carbohydrate hydrolyzing enzyme inhibition.

Cinnamomum cassia (cinnamon) proanthocyanidins (PACs) are believed to have anti-hyperglycemic potential via stimulation of insulin sensitivity. The present study investigates the carbohydrate hydrolyzing enzyme inhibition of cinnamon PACs. Five grams of cinnamon bark powder were extracted in 100 mL acetone solution (CAE) (acetone: water: hydrochloric acid, 70:29.9:0.01) for 2 h at room temperature and in 100 mL deionized water for 30 min at 90 °C (CWE). PACs were purified from CAE using LH-20 (CAE-PAC) to be further evaluated. PAC contents were evaluated by 4-Dimethylaminocinnamaldehyde (DMAC) assay and yielded 795, 177 and 123 mg/g, for CAE-PAC, CAE and CWE respectively. The total phenolic contents of CAE and CWE were determined to be 152 and 134 mg/g respectively. All extracts were adjusted to the same PAC content (180, 90, 45 and 20 µg) and the inhibitory activity against rat α-glucosidase was determined. The CAE-PAC fraction had very low rat α-glucosidase inhibitory activity, CAE had the highest (IC50 0.474 mg/mL total phenolic (TP) basis) followed by CWE (IC50 0.697 mg/mL TP basis). The specific maltase and sucrase inhibitory activities were determined and CAE (IC50 0.38 and 0.10 mg/mL TP basis) had higher inhibition than CWE (IC50 0.74 and 0.37 mg/mL TP basis). Results suggest that the observed bioactivity is not PAC dependent and that CAE has a higher anti-hyperglycemic potential than CWE via inhibition of carbohydrate hydrolyzing enzymes.

Correlation between reproductive status and steady-state messenger ribonucleic acid levels of the Myxovirus resistance gene, MX2, in peripheral blood leukocytes of dairy heifers.

The objectives of the current study were to evaluate the correlation between reproductive status and steady-state levels of Myxovirus resistance gene (MX2) mRNA in peripheral blood leukocytes (PBL) of dairy heifers and the reliability of using change in MX2 messenger RNA (mRNA) for identification of nonpregnant heifers 18 to 19 d after AI. Holstein heifers (n = 266), 13 +/- 1 mo of age, were assigned randomly to be inseminated (BRED; n = 214) or not (NONBRED; n = 52). Estrous cycles of all heifers were synchronized with an intravaginal insert containing progesterone for 7 d. At insert removal, heifers received an injection of PGF2alpha. Heifers in the BRED group were inseminated on detection of estrus or at a fixed time, 72 h after insert removal concomitant with a GnRH treatment. Heifers in the NONBRED group received an injection of GnRH 48 h after insert removal. Blood samples collected on d 0 (d of AI or estrus) and 18 were used to determine steady-state levels of MX2 mRNA. Samples collected on d 0, 7, 14, and 21 were analyzed for progesterone concentration. Pregnancy was determined retrospectively by progesterone concentration on d 21 and was diagnosed at 30 +/- 1 and 60 +/- 3 d after AI. The fold change in levels of MX2 mRNA from d 0 to 18 was greater for heifers classified and diagnosed as pregnant on d 21 (P < 0.05) and 30 +/- 1 (P < 0.05) and 60 +/- 3 (P < 0.05) d after AI compared with nonpregnant (bred but not pregnant) and NONBRED heifers. Heifers that experienced pregnancy loss from 21 to 30 +/- 1 (P = 0.11) or 21 to 60 +/- 3 (P = 0.08) d of gestation tended to have smaller fold increases in MX2 mRNA expression than those that maintained pregnancy. The sensitivity (range 57.1 to 65.6%) and negative predictive values (range 47.9 to 57.1%) of determining reproductive status on d 18 according to the change in the level of MX2 mRNA expression in PBL were low, and the correlation between diagnosis of pregnancy by fold change in MX2 mRNA expression and other methods was small (r = 0.20 to 0.36). The current study indicates that increased expression of MX2 mRNA in PBL is related to pregnancy approximately 21, 30, and 60 d after AI in dairy heifers and that losses that occurred later in pregnancy were associated with lower fold increases in MX2 mRNA. However, using the change in MX2 mRNA expression was not a reliable method for diagnosis of pregnancy at 18 d after AI because of the low sensitivity and negative predictive value.

Regulation of interferon-stimulated genes in peripheral blood leukocytes in pregnant and bred, nonpregnant dairy cows.

In ruminants, pregnancy results in up-regulation of a large number of IFN-stimulated genes (ISG) in the uterus. Recently, one of these genes was also shown to increase in peripheral blood leukocytes (PBL) during early pregnancy in sheep. Our working hypothesis is that conceptus signaling activates maternal gene expression in PBL in dairy cattle. The objectives of this study were to characterize ISG expression in PBL from pregnant (n = 20) and bred, nonpregnant (n = 30) dairy cows. Steady-state levels of mRNA for Mx1, Mx2, beta2-microglobulin, ISG-15, IFN regulatory factor-1, and IFN regulatory factor-2 were quantified. Holstein cows were synchronized to estrus and artificially inseminated (d 0). Blood samples were collected (coccygeal venipuncture) on d 0 and 16, 18, and 20 d after insemination for progesterone analysis and PBL isolation. Pregnancy was confirmed by transrectal ultrasonography at approximately 40 d after breeding. A status x day interaction was detected for Mx1, Mx2, and ISG-15 gene expression. When analyzed within day, levels of mRNA for ISG-15 and Mx1 were greater in pregnant compared with bred, nonpregnant cows on d 18 and 20, respectively. Expression of the Mx2 gene increased in the pregnant group compared with bred, nonpregnant cows on d 16, 18, and 20 after insemination. beta2-Microglobulin, IFN regulatory factor-1, and IFN regulatory factor-2 were not different between groups. The results clearly indicated that components of the innate immune response are activated in PBL during the period of pregnancy recognition and early embryo signaling. The physiological implications of these changes on maternal immune function are as yet unknown; however, they do provide a unique opportunity to identify bred, nonpregnant, cows 18 d after insemination in dairy cattle.