Soluble MAC is primarily released from MAC-resistant bacteria that potently convert complement component C5

The Membrane Attack Complex (MAC or C5b-9) is an important effector of the immune system to kill invading microbes. MAC is formed when complement enzymes on the bacterial surface convert complement component C5 into C5b. Although the MAC is a membrane-inserted complex, soluble forms of MAC (sMAC, or terminal complement complex (TCC)) are often detected in sera of patients suffering from infections. Consequently, sMAC has been proposed as a biomarker, but it remains unclear when and how it is formed during infections. Here, we studied mechanisms of MAC formation on bacteria and found that sMAC is primarily formed in human serum by bacteria resistant to MAC-dependent killing. Surprisingly, C5 was converted into C5b more potently by MAC-resistant compared to MAC-sensitive Escherichia coli strains. Both the increase in C5 conversion and sMAC generation were linked to the expression of lipopolysaccharide (LPS) O-Antigen in the bacterial outer membrane. In addition, we found that MAC precursors are released from the surface of MAC-resistant bacteria during MAC assembly. Release of MAC precursors from bacteria induced lysis of bystander human erythrocytes in the absence of other serum components. However, serum regulators vitronectin (Vn) and clusterin (Clu) can prevent this bystander lysis. Combining size exclusion chromatography with mass spectrometry profiling, we show that sMAC released from bacteria in serum is a heterogeneous mixture of complexes composed of C5b-8, up to 3 copies of C9 and multiple copies of Vn and Clu. Altogether, our data provide molecular insight into how sMAC is generated during bacterial infections. This fundamental knowledge could form the basis for exploring the use of sMAC as biomarker.

Analysis of the Fluid Contents of Simple Bone Cyst in the Mandible: Blood or Serum?

Description of the content of simple bone cyst (SBC) has been controversial. This study aimed to evaluate and give a clearer picture of the contents of the SBC cavity. Nineteen patients SBC confirmed by histopathological examination, between 2014 and 2016, were included in this study. The clinical, radiographic, surgical and laboratory findings of SBC cavity content was analyzed. For statistical analysis, paired samples t-test was used to evaluate the difference of components among cavity fluid, blood and serum. All 19 SBC cases radiographically and surgically revealed a fluid-filled cavity. The average age of the patients was 21.3±13.2 years. Any sex predominance was not found. SBCs were more often found in the anterior mandible site (n=12, 63.2%). All lesions were filled with clear straw-colored or low concentrated blood-colored flood. Although the laboratory analysis of the fluid showed components similar to those in blood and serum, the statistical analysis showed that the fluid components were not significantly different only for eosinophils (p = 0.43) and basophils (p = 0.06) counts as blood components, and sodium (p = 0.76), potassium (p = 0.08) and chloride (p = 0.13) concentration as serum components. Within a limitation of this study, the results shows that SBC is fluid-filled cavity and the cavity fluid is more likely similar to the serum rather than the blood regarding the internal components.

Identification of novel candidate factor SNX5 specific for pregnancy failure in Holstein heifers

Pregnancy loss predominantly occurs during periods between blastocyst hatching and conceptus (embryo plus extraembryonic membranes) implantation to the endometrium in cattle. Insufficient biochemical communication between conceptus and endometrium has been suspected as the primary cause for early embryonic losses. If molecules regulating this communication were identified, molecular mechanisms associated with early pregnancy success or loss could be better understood. To identify novel factors as detection markers of non-pregnant or females undergoing embryonic loss, blood sera from embryo-transferred heifers on day 7 (day 0 = day of estrus) were collected on day 17, 20, or 22, which were subjected to metabolome and global proteome iTRAQ analyses. On each sample, the metabolome analysis partly divided serum components into pregnant or not. In the iTRAQ analysis, heatmap analysis with 25 unique proteins separated into pregnant or not on day 20 or 22. Furthermore, receiver operating characteristic curve (ROC) analysis identified five candidate proteins detecting non-pregnant heifers, of which SNX5 in day 22 sera had the highest area under the curve (AUC), 0.983. We also detected SNX5 in day 22 sera from non-pregnant heifers using western blotting. These results suggest that high SNX5 in day 22 sera could predict early pregnancy loss in heifers.

A total number of 192 male one-day-old broilers chickens were randomly divided into four treatment groups of 48 chickens. Chickens of group one fed a plain diet without any supplement (control), while the diets ib groups two, three, and four were supplemented with Whole Yeast (WY, Saccharomyces cerevisiae, 0.1%), Yeast Cell Wall (YCW, 0.3 %), and Yeast Extract (YE, 0.07 %), respectively. At the end of the experimental period (35 days), the bodyweight of chickens and the feed intake of each cage were measured, and then the feed conversation ratio was calculated. Blood samples were also collected to measure the serum components and relative spleen, bursa of Fabricius, and thymus gland. The results obtained indicated that all productive performance parameters improved in response to the feeding supplementation. Blood parameters indicated that the treated groups had a significantly higher level of serum total protein and albumin as well as significantly lower serum total lipids and cholesterol. The enzyme activities of ALT, AST, and ALP were significantly reduced by WY, YCW, and YE supplementation. The relative organ weights of the spleen, bursa of Fabricius, and thymus increased significantly in broilers fed with WY, YCW, and YE, and the highest values were observed in the chickens fed with WY. It can be demonstrated that the supplementation of WY or its derivatives in the diet of broiler chickens improves the production performance as well as the physiological and immunological parameters, and consequently produce a healthier chicken

A total number of 192 male one-day-old broilers chickens were randomly divided into four treatment groups of 48 chickens. Chickens of group one fed a plain diet without any supplement (control), while the diets ib groups two, three, and four were supplemented with Whole Yeast (WY, Saccharomyces cerevisiae, 0.1%), Yeast Cell Wall (YCW, 0.3 %), and Yeast Extract (YE, 0.07 %), respectively. At the end of the experimental period (35 days), the bodyweight of chickens and the feed intake of each cage were measured, and then the feed conversation ratio was calculated. Blood samples were also collected to measure the serum components and relative spleen, bursa of Fabricius, and thymus gland. The results obtained indicated that all productive performance parameters improved in response to the feeding supplementation. Blood parameters indicated that the treated groups had a significantly higher level of serum total protein and albumin as well as significantly lower serum total lipids and cholesterol. The enzyme activities of ALT, AST, and ALP were significantly reduced by WY, YCW, and YE supplementation. The relative organ weights of the spleen, bursa of Fabricius, and thymus increased significantly in broilers fed with WY, YCW, and YE, and the highest values were observed in the chickens fed with WY. It can be demonstrated that the supplementation of WY or its derivatives in the diet of broiler chickens improves the production performance as well as the physiological and immunological parameters, and consequently produce a healthier chicken.

Metabolic plasticity enables lifestyle transitions of Porphyromonas gingivalis

Our understanding of how the oral anaerobe Porphyromonas gingivalis can persist below the gum line, induce ecological changes, and promote polymicrobial infections remains limited. P. gingivalis has long been described as a highly proteolytic and asaccharolytic pathogen that utilizes protein substrates as the main source for energy production and proliferation. Here, we report that P. gingivalis displays a metabolic plasticity that enables the exploitation of non-proteinaceous substrates, specifically the monocarboxylates pyruvate and lactate, as well as human serum components, for colonization and biofilm formation. We show that anabolism of carbohydrates from pyruvate is powered by catabolism of amino acids. Concomitantly, the expression of fimbrial adhesion is upregulated, leading to the enhancement of biofilm formation, stimulation of multispecies biofilm development, and increase of colonization and invasion of the primary gingival epithelial cells by P. gingivalis. These studies provide the first glimpse into the metabolic plasticity of P. gingivalis and its adaptation to the nutritional condition of the host niche. Our findings support the model that in response to specific nutritional parameters, P. gingivalis has the potential to promote host colonization and development of a pathogenic community.

Reference Intervals in Combined Veterinary Clinical Examinations of Male Black-Spotted Pond Frogs (Pelophylax nigromaculatus)

In conservation physiology, analyzing the physiological response of an organism to understand its ability to adapt to environmental changes is a key technique in establishing a successful conservation strategy. Veterinary clinical examinations determine the physiological condition of animals accurately and safely, and this examination is synergistic when combined. The accuracy and safety of a clinical examination makes it advantageous for use in amphibians with high species diversity and numerous endangered species. However, it is necessary to establish a reference interval (RI) for precise interpretations and identification of animals with abnormalities through individual unit testing. We have established RIs for the immunity, serum components, bone mineral density (BMD), and body composition of black-spotted pond frogs (Pelophylax nigromaculatus). Black-spotted pond frogs are a common species and are widely distributed in East Asia, with suitable characteristics for environmental monitoring. Serum was extracted from 151 male frogs to establish the RI for bacterial killing ability in order to represent immunity. We also used the serum to establish an RI of ten additional serum components to determine the nutritional status, organ function status, body osmotic pressure, and homeostasis conditions. The BMD and three body composition measures for diagnosing food intake status and nutritional condition were measured using dual energy X-ray absorptiometry. The RI was recorded as the mean ± standard deviation, median, first (25%) to third (75%) quantile range, 95% confidence interval of the mean and median, and the 95% percentile (2.5%–97.5% range) of all components. The use of combined clinical veterinary examinations aids our understanding of the physiological conditions of an individual according to biotic and abiotic factors on a complex spatiotemporal scale in an ecosystem.

LPA signaling acts as a cell-extrinsic mechanism to initiate cilia disassembly and promote neurogenesis

Dynamic assembly and disassembly of primary cilia controls embryonic development and tissue homeostasis. Dysregulation of ciliogenesis causes human developmental diseases termed ciliopathies. Cell-intrinsic regulatory mechanisms of cilia disassembly have been well-studied. The extracellular cues controlling cilia disassembly remain elusive, however. Here, we show that lysophosphatidic acid (LPA), a multifunctional bioactive phospholipid, acts as a physiological extracellular factor to initiate cilia disassembly and promote neurogenesis. Through systematic analysis of serum components, we identify a small molecular—LPA as the major driver of cilia disassembly. Genetic inactivation and pharmacological inhibition of LPA receptor 1 (LPAR1) abrogate cilia disassembly triggered by serum. The LPA-LPAR-G-protein pathway promotes the transcription and phosphorylation of cilia disassembly factors-Aurora A, through activating the transcription coactivators YAP/TAZ and calcium/CaM pathway, respectively. Deletion of Lpar1 in mice causes abnormally elongated cilia and decreased proliferation in neural progenitor cells, thereby resulting in defective neurogenesis. Collectively, our findings establish LPA as a physiological initiator of cilia disassembly and suggest targeting the metabolism of LPA and the LPA pathway as potential therapies for diseases with dysfunctional ciliogenesis.

Effects of Improved (Addition of Antimicrobials and Antioxidants) Locally Produced Natural Vitamin Premix on Performance, Hematology and Some Serum Constituents of Broiler Chickens

An experiment was conducted to compare the haematology and some serum constituents and performance of broilers fed diets containing improved (addition of anti-microbial and anti-oxidant) locally produced natural vitamin premix (LPNVP), LPNVP without improvement and commercial vitamin premix (CVMP). Haematology of the birds was not affected by the dietary treatments. However, red blood cell (RBC) count was significantly high (P<0.05) for the birds fed diet containing improved LPNVP The haemoglobin, PCK, RBC and WBC values observed for birds fed improved LPNVP were 13 gm%, 32%, 246 x 1012/L and 3.1 x 1010/L respectively. Serological parameters observed in the study were not significantly different (P>0,.05) for all the dietary conditions studied. Birds fed diet containing improved LPNVP also recorded comparable serum components to control diet containing the commercial vitamin/’mineral premix. Serum calcium, phosphorus, glucose, ureate, creatinine and total protein observed for birds fed diet containing improved LPNVP were 2.32%, 0.70%, 3.60Mmo/L, 2.9Mmo/L, 0.25Mmol/L, 58Mmol/L and 66g/100ml respectively. Feed intake, weight gain and feed utilization were similar (P>0.05) for the various dietary treatments. Birds fed improved LPNVP based diets tended to record better feed intake than did the control birds on CVMP based diets and unimproved LPNVP. Generally, improvement of LPNVP tended to give better performance of broilers than did other dietary treatments.