Cytokines in \u3ci\u3eMycoplasma hyorhinis\u3c/i\u3e-Induced Arthritis in Pigs Bred Selectively for High and Low Immune Responses

Yorkshire pigs were bred selectively for high and low immune responses (H and L pigs, respectively) based on multiple antibody (Ab) and cell-mediated immune response traits. In a previous experiment, generation 4 (G4) pigs of each line were infected with Mycoplasma hyorhinis. High responders had a more rapid and higher Ab response and less polyserositis, but arthritis was more severe in H pigs than in L pigs. To test the hypothesis that line differences were attributable to differential expression of cytokines, M. hyorhinis infection was induced in pigs of G8. Arthritis was more severe clinically (P, ≤0.05) and postmortem (P, ≤0.001) when M. hyorhinis CFU were more numerous in synovial fluid (SF) of H pigs than of L pigs (P, ≤0.03). In H pigs but not L pigs, CFU and lesion scores were correlated positively. In H pigs, infection increased the frequency of expression of mRNAs for interleukin-8 (IL-8), IL-10, and tumor necrosis factor alpha (TNF-α) in mononuclear cells from synovial membranes (SM). In L pigs, IL-1a, IL-6, IL-10, and TNF-a mRNAs were increased in frequency of expression. The quantity of the cytokine message for IL-6 was increased in infected H pigs. For L pigs, infection increased the cytokine message for IL-1 α, IL-6, IL-10, and TNF-a. IL-6 in SM and gamma interferon (IFN-ϒ) in SF were produced at a higher copy number in H pigs than in L pigs after infection. For H pigs, there were no positive rank correlations between lesion or CFU scores and cytokines. For L pigs, IL-1 α, IL-8, IL-10, and TNF- α in SM correlated with CFU, while IL-6, TNF- β, and IFN-ϒ in SF correlated with CFU. Lesion score in L pigs correlated with IL-1 α in SF. While these results indicate that H and L pigs differ in the cytokine response to M. hyorhinis infection, they do not confirm a characteristic cytokine response in association with the relative susceptibility to infection and arthritis observed in H pigs

Cytokines in Pigs Bred Selectively for High and Low Immune Response [abstract only]

Yorkshire pigs have been bred for high (H) and low (L) immune response based on selection for multiple antibody (Ab) and cell mediated immune response traits. High responders have better production and larger litter size when compared with controls and low responders. The ability of high and low line pigs to resist M. hyorhinis infection has been tested. The high responders had more rapid and higher Ab response and the severity of the disease was less, as judged by clinical and postmortem signs. However, arthritis was found to be relatively more severe in high responders. We hypothesized that the immune response differences between genetically different lines could be attributed to either dominant or differential cytokine expression. To test the above hypothesis, quantitative RNA PCR (Q.RNA PCR), to quantitate the porcine cytokines at the mRNA level, was developed by constructing an internal control. Two synthetic oligos, namely 5\u27 construct (FPC) and 3\u27 construct (TPC), were designed based on the nt sequences of porcine cytokine genes. FPC represented the upstream primer sequences of nine cytokines sequentially in the order IL-1, IL-4, IL-6, IL-8, IL-2, IL-1O, TNF-α, TNF-β and IFN-γ, and TPC, the downstream primer sequences in the same order. The primers were designed such that when cRNA and target RNA were amplified, they give two non-overlapping products. FPCs and TPCs were constructed by overlapping and the extension method of PCR amplification utilizing six oligos for each, and were cloned into pSP 64 poly A vector. The application of Q.RNA PCR has been tested for determining quantitatively the cytokines in peripheral blood mononuclear cells in H-L line pigs. Preliminary study indicated differential expression of cytokines, namely IL-1, IL-6, IL-1O, TNF-α and IFN-γ, in naive animals. Expression of other cytokines, namely IL-2, IL-4, IL-8 and TNF-β, was absent in the pigs tested. Future studies involve the determination of cytokines in the context of immunization to the antigens (HEWL, BCG, etc.) as well as during infection (ex: M. hyorhinis) in conjunction with cytokine expression regulation strategies, namely MAbs and/or antisense ODNs or gene therapy

Voltage Regulator Module Noise Analysis for High-Volume Server Applications

This paper presents a methodology to analyze voltage regulator module (VRM) noise coupling problems in high-volume server applications. The technique is applied on a real engineering design. The comprehensive model includes irregular power shapes, decoupling capacitors, and dielectric and conductive loss. Irregular shaped power plane modeling is cross-checked with four separate methods to demonstrate accuracy

The Ups and Downs of Mutation Frequencies during Aging Can Account for the Apert Syndrome Paternal Age Effect

Apert syndrome is almost always caused by a spontaneous mutation of paternal origin in one of two nucleotides in the fibroblast growth factor receptor 2 gene (FGFR2). The incidence of this disease increases with the age of the father (paternal age effect), and this increase is greater than what would be expected based on the greater number of germ-line divisions in older men. We use a highly sensitive PCR assay to measure the frequencies of the two causal mutations in the sperm of over 300 normal donors with a wide range of ages. The mutation frequencies increase with the age of the sperm donors, and this increase is consistent with the increase in the incidence rate. In both the sperm data and the birth data, the increase is non-monotonic. Further, after normalizing for age, the two Apert syndrome mutation frequencies are correlated within individual sperm donors. We consider a mathematical model for germ-line mutation which reproduces many of the attributes of the data. This model, with other evidence, suggests that part of the increase in both the sperm data and the birth data is due to selection for mutated premeiotic cells. It is likely that a number of other genetic diseases have similar features

Convalescent plasma in patients admitted to hospital with COVID-19 (RECOVERY): a randomised controlled, open-label, platform trial

Background: Many patients with COVID-19 have been treated with plasma containing anti-SARS-CoV-2 antibodies. We aimed to evaluate the safety and efficacy of convalescent plasma therapy in patients admitted to hospital with COVID-19. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]) is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. The trial is underway at 177 NHS hospitals from across the UK. Eligible and consenting patients were randomly assigned (1:1) to receive either usual care alone (usual care group) or usual care plus high-titre convalescent plasma (convalescent plasma group). The primary outcome was 28-day mortality, analysed on an intention-to-treat basis. The trial is registered with ISRCTN, 50189673, and ClinicalTrials.gov, NCT04381936. Findings: Between May 28, 2020, and Jan 15, 2021, 11558 (71%) of 16287 patients enrolled in RECOVERY were eligible to receive convalescent plasma and were assigned to either the convalescent plasma group or the usual care group. There was no significant difference in 28-day mortality between the two groups: 1399 (24%) of 5795 patients in the convalescent plasma group and 1408 (24%) of 5763 patients in the usual care group died within 28 days (rate ratio 1·00, 95% CI 0·93–1·07; p=0·95). The 28-day mortality rate ratio was similar in all prespecified subgroups of patients, including in those patients without detectable SARS-CoV-2 antibodies at randomisation. Allocation to convalescent plasma had no significant effect on the proportion of patients discharged from hospital within 28 days (3832 [66%] patients in the convalescent plasma group vs 3822 [66%] patients in the usual care group; rate ratio 0·99, 95% CI 0·94–1·03; p=0·57). Among those not on invasive mechanical ventilation at randomisation, there was no significant difference in the proportion of patients meeting the composite endpoint of progression to invasive mechanical ventilation or death (1568 [29%] of 5493 patients in the convalescent plasma group vs 1568 [29%] of 5448 patients in the usual care group; rate ratio 0·99, 95% CI 0·93–1·05; p=0·79). Interpretation: In patients hospitalised with COVID-19, high-titre convalescent plasma did not improve survival or other prespecified clinical outcomes. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research

Tocilizumab in patients admitted to hospital with COVID-19 (RECOVERY): a randomised, controlled, open-label, platform trial

Background: In this study, we aimed to evaluate the effects of tocilizumab in adult patients admitted to hospital with COVID-19 with both hypoxia and systemic inflammation. Methods: This randomised, controlled, open-label, platform trial (Randomised Evaluation of COVID-19 Therapy [RECOVERY]), is assessing several possible treatments in patients hospitalised with COVID-19 in the UK. Those trial participants with hypoxia (oxygen saturation <92% on air or requiring oxygen therapy) and evidence of systemic inflammation (C-reactive protein ≥75 mg/L) were eligible for random assignment in a 1:1 ratio to usual standard of care alone versus usual standard of care plus tocilizumab at a dose of 400 mg–800 mg (depending on weight) given intravenously. A second dose could be given 12–24 h later if the patient's condition had not improved. The primary outcome was 28-day mortality, assessed in the intention-to-treat population. The trial is registered with ISRCTN (50189673) and ClinicalTrials.gov (NCT04381936). Findings: Between April 23, 2020, and Jan 24, 2021, 4116 adults of 21 550 patients enrolled into the RECOVERY trial were included in the assessment of tocilizumab, including 3385 (82%) patients receiving systemic corticosteroids. Overall, 621 (31%) of the 2022 patients allocated tocilizumab and 729 (35%) of the 2094 patients allocated to usual care died within 28 days (rate ratio 0·85; 95% CI 0·76–0·94; p=0·0028). Consistent results were seen in all prespecified subgroups of patients, including those receiving systemic corticosteroids. Patients allocated to tocilizumab were more likely to be discharged from hospital within 28 days (57% vs 50%; rate ratio 1·22; 1·12–1·33; p<0·0001). Among those not receiving invasive mechanical ventilation at baseline, patients allocated tocilizumab were less likely to reach the composite endpoint of invasive mechanical ventilation or death (35% vs 42%; risk ratio 0·84; 95% CI 0·77–0·92; p<0·0001). Interpretation: In hospitalised COVID-19 patients with hypoxia and systemic inflammation, tocilizumab improved survival and other clinical outcomes. These benefits were seen regardless of the amount of respiratory support and were additional to the benefits of systemic corticosteroids. Funding: UK Research and Innovation (Medical Research Council) and National Institute of Health Research

Hepatic Protoporphyria in Weanling Pigs

Hepato-cutaneous porphyria can be induced in weanling pigs. The collidine derivative, DDC, produces protoporphyria in these animals, associated with photosensitivity. This porphyria is characterized by accumulation of protoporphyrin in the bile and liver. Fecal protoporphyrin excretion is markedly increased. The levels of protoprophyrin in the bile and liver are dependent on the dosage level of DDC. Erythrocyte protoporphyrin levels are not elevated. Protoporphyrin in the plasma and skin of animals on the maximal DDC dose was demonstrated. There was no significant elevation of urinary porphyrins in the experimental pigs