Milk Fat Depression and Trans-11 to Trans-10 C18:1 Shift in Milk of Two Cattle Farming Systems

Milk fat depression (MFD) syndrome, a consistent decrease in milk fat content, is related to important changes in fatty acid composition due to feed imbalances and the consequent ruminal metabolism alteration. Milk produced in two different farming systems was compared: Holstein Friesian fed with unified in intensive production and Podolica raised on a pasture in an extensive system. Milk chemical characteristics and fatty acid composition were determined comparing milk with a normal fat level (>3.8%) to milk with a low fat level (<3.2%) in each breeding system. Holstein Friesian milk showed the decrease in trans-11 and increase in trans-10 C18:1 (shift from trans-11 to trans-10 C18:1) in low fat with respect to normal fat milk with a consequent decrease in the trans-11/trans-10 C18:1 ratio. Even conjugated linoleic acid (CLA), C18:2 cis-9, trans-11, was lower while CLA trans-10, cis-12 was higher in low fat milk than in normal fat milk from Holstein Friesian. These changes, that are indicators of MFD syndrome, were not found in Podolica milk between fat levels. Holstein Friesian milk showed less short-chain fatty acids (9.48 % vs. 11.05%, p < 0.001), trans vaccenic acid (C18:1 trans-11, 0.51% vs. 3.39%, p < 0.001), rumenic acid (CLA C18:2 cis-9, trans-11, 0.32% vs. 1.45%, p < 0.001) and total CLA (0.53% vs. 1.91%, p < 0.001) contents than Podolica milk. Further losses of these human healthy nutrients in low fat Friesian milk reduced the nutritional quality of the milk, while the milk from animals raised on the pasture was of better quality even when the level of fat was low.

Whole Exome Sequencing detects PYGM variants in two adults with McArdle disease

McArdle disease is a progressive and debilitating glycogen storage disease with typical onset in late childhood. Here we describe a former competitive athlete with early adult onset McArdle disease and a septuagenarian with a history of exercise-intolerance since adolescence who was evaluated for proximal muscle weakness. Exome sequencing identified bi-allelic variants in PYGM gene for both cases. The former athlete has the common, well-known pathogenic variant p.(Arg50Ter) in trans with a novel missense variant, p.(Asp694Glu). The second individual has a previously described homozygous missense variant, p.(Arg771Gln). Here, we describe the clinical course, enzyme-testing results using muscle tissue and molecular findings for the individuals, and add to the knowledge of the genotypic spectrum of this disorder.

Structural basis of BAK activation in mitochondrial apoptosis initiation

BCL-2 proteins regulate mitochondrial poration in apoptosis initiation. How the pore-forming BCL-2 Effector BAK is activated remains incompletely understood mechanistically. Here we investigate autoactivation and direct activation by BH3-only proteins, which cooperate to lower BAK threshold in membrane poration and apoptosis initiation. We define in trans BAK autoactivation as the asymmetric “BH3-in-groove” triggering of dormant BAK by active BAK. BAK autoactivation is mechanistically similar to direct activation. The structure of autoactivated BAK BH3-BAK complex reveals the conformational changes leading to helix α1 destabilization, which is a hallmark of BAK activation. Helix α1 is destabilized and restabilized in structures of BAK engaged by rationally designed, high-affinity activating and inactivating BID-like BH3 ligands, respectively. Altogether our data support the long-standing hit-and-run mechanism of BAK activation by transient binding of BH3-only proteins, demonstrating that BH3-induced structural changes are more important in BAK activation than BH3 ligand affinity.

LRRC15 suppresses SARS-CoV-2 infection and controls collagen production

Although ACE2 is the primary receptor for SARS-CoV-2 infection, a systematic assessment of factors controlling SARS-CoV-2 host interactions has not been described. Here we used whole genome CRISPR activation to identify host factors controlling SARS-CoV-2 Spike binding. The top hit was a Toll-like receptor-related cell surface receptor called leucine-rich repeat-containing protein 15 (LRRC15). LRRC15 expression was sufficient to promote SARS-CoV-2 Spike binding where it forms a cell surface complex with LRRC15 but does not support infection. Instead, LRRC15 functioned as a negative receptor suppressing both pseudotyped and live SARS-CoV-2 infection. LRRC15 is expressed in collagen-producing lung myofibroblasts where it can sequester virus and reduce infection in trans. Mechanistically LRRC15 is regulated by TGF-β, where moderate LRRC15 expression drives collagen production but high levels suppress it, revealing a novel lung fibrosis feedback circuit. Overall, LRRC15 is a master regulator of SARS-CoV-2, suppressing infection and controlling collagen production associated with “long-haul” COVID-19.

Role of AmpC-Inducing Genes in Modulating Other Serine Beta-Lactamases in Escherichia coli

The consistently mutating bacterial genotypes appear to have accelerated the global challenge with antimicrobial resistance (AMR); it is therefore timely to investigate certain less-explored fields of targeting AMR mechanisms in bacterial pathogens. One of such areas is beta-lactamase (BLA) induction that can provide us with a collection of prospective therapeutic targets. The key genes (ampD, ampE and ampG) to which the AmpC induction mechanism is linked are also involved in regulating the production of fragmented muropeptides generated during cell-wall peptidoglycan recycling. Although the involvement of these genes in inducing class C BLAs is apparent, their effect on serine beta-lactamase (serine-BLA) induction is little known. Here, by using ∆ampD and ∆ampE mutants of E. coli, we attempted to elucidate the effects of ampD and ampE on the expression of serine-BLAs originating from Enterobacteriaceae, viz., CTX-M-15, TEM-1 and OXA-2. Results show that cefotaxime is the preferred inducer for CTX-M-15 and amoxicillin for TEM-1, whereas oxacillin for OXA-2. Surprisingly, exogenous BLA expressions are elevated in ∆ampD and ∆ampE mutants but do not always alter their beta-lactam susceptibility. Moreover, the beta-lactam resistance is increased upon in trans expression of ampD, whereas the same is decreased upon ampE expression, indicating a differential effect of ampD and ampE overexpression. In a nutshell, depending on the BLA, AmpD amidase moderately facilitates a varying level of serine-BLA expression whereas AmpE transporter acts likely as a negative regulator of serine-BLA.

Bradyrhizobium japonicum FN1 produces an inhibitory substance that affects competition for nodule occupancy

Bacteriocins are narrow spectrum antibiotics of bacterial origin that can affect competition in resource-limited environments such as the rhizosphere. Therefore, bacteriocins may be good candidates for manipulation in order to generate more competitive inocula for soybean. In this study,<i> B. japonicum</i> FN1 along with other Bradyrhizobia in our culture collection were screened for bacteriocin-like activity. A total of five distinct inhibitory activities could be observed. FN1 genes putatively involved in bacteriocin production were computationally identified. These genes were mutagenized and the subsequent strains were screened for loss of inhibitory activity. Mutant strain BRJ-48, with an insert in<i> bjfn1_01204</i>, displayed a loss of the ability to inhibit an indicator strain. This loss could be complemented by the introduction of a plasmid expressing <i>bjfn1_01204 </i>in trans. The strain carrying the mutation did not affect competition in broth cultures, but was shown to be less competitive for nodule occupancy. Annotation suggests that <i>bjfn1_01204</i> encodes a carboxymuconolactone decarboxylase, however the direct contribution of how this enzyme contributes to inhibiting the tester strain remains unknown.

Organizational, Technical and Behavioral Factors Associated with Malaria Routine Data Reporting among Health Workers in Selected Health Facilities in Trans-Nzoia County, Kenya

Purpose: Malaria remains to be among the primary causes of sickness, infirmity and cases of deaths and has continued to negatively affect health and socio-economic progress in the country. Rapid reporting of malaria cases could avert prospective epidemics which would lead to a high proportion of sickness and deaths. The study, therefore, sought to assess the determinants of malaria routine data reporting among health workers in selected health facilities in Trans-Nzoia County. Methodology: A descriptive cross-sectional study was conducted to evaluate the organizational, technical, and behavioral aspects that influence the reporting of malaria routine data among health workers. The sample size was 123 health facilities that were selected randomly based on their strata. Research tools that were utilized were structured questionnaires, focused group discussion, and key informant interview guide. Chi-square (χ2) was used to test the hypothesis with a p ≤ 0.05 being considered significant. Findings: The findings on socio-demographic characteristics indicated that majority of the research participants were females 76(62.6%), had college education 85(69.1%) and 81 (65.9%) had worked in the health facility for 5 to 10 years. Most 76(61.8%) of the health facilities were Level 3 (Health Centres). There was significant relationship between level of health facility and malaria routine data reporting at (χ2 =9.999, df=3, p-value = 0.019). Other organizational factors that had significant association with malaria routine data reporting (p< 0.001) include inadequate budget, low staffing, poor ICT infrastructure and complex data management procedures. In terms of technical aspects, limited training on technologies had significant relationship with malaria routine data reporting (p< 0.001). Regarding behavioral aspects, identified factors include lack of incentives and inadequate resources. Unique contribution to theory, practice and policy: The outcomes of the study provide proof for support, tactical organization, and collaboration in the health sector in Trans-Nzoia County as well as to the other developmental agencies working in the field of malaria control. The study recommends that the county government of Trans-Nzoia should provide adequate funds and ICT infrastructure to boost malaria routine data reporting. The county department of health with support from the national government through Division of National Malaria Program (DNMP )should consistently conduct in-service training, support supervision and data quality audits.