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Arginine-ornithine metabolism plays a crucial role in bacterial homeostasis, as evidenced by numerous studies. However, the utilization of arginine and the downstream products of its metabolism remain undefined in various gut bacteria. To bridge this knowledge gap, we employed genomic screening to pinpoint relevant metabolic targets. We also devised a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics method to measure the levels of arginine, its upstream precursors, and downstream products in cell-free conditioned media from enteric pathobionts, including Escherichia coli, Klebsiella aerogenes, K. pneumoniae, Pseudomonas fluorescens, Acinetobacter baumannii, Streptococcus agalactiae, Staphylococcus epidermidis, S. aureus, and Enterococcus faecalis. Our findings revealed that all selected bacterial strains consumed glutamine, glutamate, and arginine, and produced citrulline, ornithine, and GABA in our chemically defined medium. Additionally, E. coli, K. pneumoniae, K. aerogenes, and P. fluorescens were found to convert arginine to agmatine and produce putrescine. Interestingly, arginine supplementation promoted biofilm formation in K. pneumoniae, while ornithine supplementation enhanced biofilm formation in S. epidermidis. These findings offer a comprehensive insight into arginine-ornithine metabolism in enteric pathobionts., (© 2024 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.)

Borrelial pathogens are vector-borne etiological agents known to cause Lyme disease, relapsing fever, and Borrelia miyamotoi disease. These spirochetes each encode several surface-localized lipoproteins that bind components of the human complement system to evade host immunity. One borrelial lipoprotein, BBK32, protects the Lyme disease spirochete from complement-mediated attack via an alpha helical C-terminal domain that interacts directly with the initiating protease of the classical complement pathway, C1r. In addition, the B. miyamotoi BBK32 orthologs FbpA and FbpB also inhibit C1r, albeit via distinct recognition mechanisms. The C1r-inhibitory activities of a third ortholog termed FbpC, which is found exclusively in relapsing fever-causing spirochetes, remains unknown. Here, we report the crystal structure of the C-terminal domain of Borrelia hermsii FbpC to a limiting resolution of 1.5 Å. We used surface plasmon resonance and assays of complement function to demonstrate that FbpC retains potent BBK32-like anticomplement activities. Based on the structure of FbpC, we hypothesized that conformational dynamics of the complement inhibitory domains of borrelial C1r inhibitors may differ. To test this, we utilized the crystal structures of the C-terminal domains of BBK32, FbpA, FbpB, and FbpC to carry out molecular dynamics simulations, which revealed borrelial C1r inhibitors adopt energetically favored open and closed states defined by two functionally critical regions. Taken together, these results advance our understanding of how protein dynamics contribute to the function of bacterial immune evasion proteins and reveal a surprising plasticity in the structures of borrelial C1r inhibitors., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Anti-platelet factor 4 (PF4)/heparin immune complexes that cause heparin-induced thrombocytopenia (HIT) activate complement via the classical pathway. Previous studies have shown that the alternative pathway of complement substantially amplifies the classical pathway of complement activation through the C3b feedback cycle., Objectives: These studies sought to examine the contributions of the alternative pathway to complement activation by HIT antibodies., Methods: Using IgG monoclonal (KKO) and/or patient-derived HIT antibodies, we compared the effects of classical pathway (BBK32 and C1-esterase inhibitor [C1-INH]), alternative pathway (anti-factor B [fB] or factor D [fD] inhibitor) or combined classical and alternative pathway inhibition (soluble complement receptor 1 [sCR1]) in whole blood or plasma., Results: Classical pathway inhibitors BBK32 and C1-INH and the combined classical/alternative pathway inhibitor sCR1 prevented KKO/HIT immune complex-induced complement activation, including release of C3 and C5 activation products, binding of immune complexes to B cells, and neutrophil activation. The alternative pathway inhibitors fB and fD, however, did not affect complement activation by KKO/HIT immune complexes. Similarly, alternative pathway inhibition had no effect on complement activation by unrelated immune complexes consisting of anti-dinitrophenyl (DNP) antibody and the multivalent DNP--keyhole limpet hemocyanin antigen., Conclusions: Collectively, these findings suggest the alternative pathway contributes little in support of complement activation by HIT immune complexes. Additional in vitro and in vivo studies are required to examine if this property is shared by most IgG-containing immune complexes or if predominance of the classic pathway is limited to immune complexes composed of multivalent antigens., (© 2022 International Society on Thrombosis and Haemostasis.)

Pathogens that traffic in the blood of their hosts must employ mechanisms to evade the host innate immune system, including the complement cascade. The Lyme disease spirochete, Borreliella burgdorferi , has evolved numerous outer membrane lipoproteins that interact directly with host proteins. Compared to Lyme disease-associated spirochetes, relatively little is known about how an emerging tick-borne spirochetal pathogen, Borrelia miyamotoi , utilizes surface lipoproteins to interact with a human host. B. burgdorferi expresses the multifunctional lipoprotein, BBK32, that inhibits the classical pathway of complement through interaction with the initiating protease C1r, and also interacts with fibronectin using a separate intrinsically disordered domain. B. miyamotoi encodes two separate bbk32 orthologs denoted fbpA and fbpB ; however, the activities of these proteins are unknown. Here, we show that B. miyamotoi FbpA binds human fibronectin in a manner similar to B. burgdorferi BBK32, whereas FbpB does not. FbpA and FbpB both bind human complement C1r and protect a serum-sensitive B. burgdorferi strain from complement-mediated killing, but surprisingly, differ in their ability to recognize activated C1r versus zymogen states of C1r. To better understand the observed differences in C1r recognition and inhibition properties, high-resolution X-ray crystallography structures were solved of the C1r-binding regions of B. miyamotoi FbpA and FbpB at 1.9Å and 2.1Å, respectively. Collectively, these data suggest that FbpA and FbpB have partially overlapping functions but are functionally and structurally distinct. The data presented herein enhances our overall understanding of how bloodborne pathogens interact with fibronectin and modulate the complement system., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Booth, Powell-Pierce, Skare and Garcia.)

Ultraviolet radiation (UVR) plays a key role in several biological functions, including human health. Skin exposure to UVR is the main factor in vitamin D photoconversion. There is also evidence relating low levels of vitamin D with certain internal cancers, mainly colon, breast and prostate, as well as other diseases. Several epidemiological studies have shown an inverse relationship between the above-mentioned diseases and latitude, in accordance with the ultraviolet radiation latitudinal gradient. The aim of this study is to determine whether UV irradiance levels in the southern South America are sufficient to produce suitable levels of vitamin D year around. For this purpose, vitamin D photoconversion weighted-irradiance was analyzed between S.S. de Jujuy (24.17°S, 65.02°W) and Ushuaia (54° 50'S, 68° 18'W). In addition to irradiance, skin type and area of body exposed to sunlight are critical factors in vitamin D epidemiology. Due to a broad ethnic variability, it was assumed that the skin type in this region varies between II and V (from the most to the less sensitive). All sites except South Patagonia indicate that skin II under any condition of body area exposure and skin V when exposing head, hands, arms and legs, would produce suitable levels of vitamin D year round (except for some days in winter at North Patagonian sites). At South Patagonian sites, minimum healthy levels of vitamin D year round can be reached only by the more sensitive skin II type, if exposing head, hands, arms and legs, which is not a realistic scenario during winter. At these southern latitudes, healthy vitamin D levels would not be obtained between mid May and beginning of August if exposing only the head. Skin V with head exposure is the most critical situation; with the exception of the tropics, sun exposure would not produce suitable levels of vitamin D around winter, during a time period that varies with latitude. Analyzing the best exposure time during the day in order to obtain a suitable level of vitamin D without risk of sunburn, it was concluded that noon is best during winter, as determined previously. For skin type II when exposing head, exposure period in winter varies between 30 and 130 min, according to latitude, except for South Patagonian sites. During summer, noon seems to be a good time of day for short periods of exposure, while during leisure times, longer periods of exposure without risk of sunburn are possible at mid-morning and mid-afternoon. At 3 h from noon, solar zenith angles are almost the same for sites between the tropics and North Patagonia, and at 4 h from noon, for all sites. Then, in these cases, the necessary exposure periods varied slightly between sites, only due to meteorological differences.

The exposure of organisms to ultraviolet radiation (UVR) is characterized by the climatology (annual cycle) and the variance (anomalies) of biologically-weighted irradiances at eight geographical locations in austral South America, from 1995-2002. The net effect of UVR on biological systems is a result of the balance of damage and repair which depends on intensity and duration of irradiance and is modulated by its variability. The emphasis in this study is on day-to-day variability, a time scale of importance to adaptive strategies that counteract UVR damage. The irradiances were weighted with DNA- and phytoplankton photosynthesis-action spectra. Low latitude sites show high average UVR. For all sites, the frequency of days with above average irradiances is higher than below average irradiances. Persistence in anomalies is generally low (

The error in irradiance measured with Sun-calibrated multichannel radiometers may be large when the solar zenith angle (SZA) increases. This could be particularly detrimental in radiometers installed at mid and high latitudes, where SZAs at noon are larger than 50 degrees during part of the year. When a multiregressive methodology, including the total ozone column and SZA, was applied in the calculation of the calibration constant, an important improvement was observed. By combining two different equations, an improvement was obtained at almost all the SZAs in the calibration. An independent test that compared the irradiance of a multichannel instrument and a spectroradiometer installed in Ushuaia, Argentina, was used to confirm the results.

Hematocrit (Hct), hemoglobin concentration, RBC count, and blood parasite loads (Plasmodium, Leucocytozoon, Haemoproteus, Trypanosoma, microfilariae) were measured in 688 passerine birds captured at four sites in the US and Belize, C.A., to assess physiological consequences of parasitemia. Parasite loads were low overall: 80% of birds had <5 parasites/10000 RBCs; median infection intensity was 1 per 10000 RBCs. Infection prevalence and intensity were higher in Belize than in the US. Analyses across and within taxa revealed no significant linear correlations between hematology values and parasite load, though infected birds were more likely to be anemic. Neotropical migrants reaching the southern US had lower parasite loads than migrants wintering in Belize. Mean Hct was significantly elevated among infected migrants (but not uninfected migrants) reaching the US relative to migrants in Belize. Long-distance migration thus appears to act as a filter for heavily parasitized and anemic birds. Geographic and seasonal variations in hematology indices were similar within and between taxa, suggesting that community-wide factors, rather than phylogenetic attributes or species-specific pathogens, account for the majority of hematological variation. The low parasite loads and weak correlation between parasite load and hematology indices have implications for attempts to test the version of the 'good genes' hypothesis.

Variations in the host response during pneumonia caused by Streptococcus pneumoniae in susceptible (CBA/Ca) and resistant (BALB/c) inbred mouse strains were investigated. Significant differences were detected in survival time, core body temperature, lung-associated and systemic bacterial loads, mast cell numbers, magnitude and location of cytokine production, lung disruption, and ability of isolated lung cells to release the cytokine tumor necrosis factor (TNF) alpha in vitro. Overall, the results indicate that the reduced capacity of CBA/Ca mice to induce rapid TNF activity within the airways following infection with S. pneumoniae may be a factor in their elevated susceptibility to pneumococcal pneumonia.

Hemolymph samples were taken from small (< 100 g) individuals of Cancer productus following ca. 3 h air exposure (emersion) on the beach, `in situ', at Friday Harbor, Washington. Compared with crabs of similar size in sea water in the laboratory, these crabs emersed `in situ' had lower Pa o 2 , and Pv o 2 , but no significant change in pH and a small, not significant, internal hypercapnia. Total CO 2 (C c o 2 ) content of the hemolymph was elevated by 70% (15.2 versus 9.0 mM), possibly as compensation for input of acid into the hemolymph. These responses are qualitatively similar to those resulting from similar treatment in the laboratory, but differ in the reduced magnitude of the internal hypercapnia and acidosis of the hemolymph. It is suggested that the particular conditions of emersion `in situ' permit some gas exchange with interstitial sea water. Interstitial sea water was found to be hypoxic (P o 2 = 20-40 torr), which would limit oxygen supply yet permit CO 2 excretion to continue, in agreement with the data.