Infected Control Group Research Articles

Polyherbal Syrup for Coronavirus Infection: Formulation and Evaluation

Background: SARS-CoV-2 emerged in Wuhan in December 2019, and after that, it spread quickly around the world. The virus could spread to millions of individuals since there were no particular treatments or preventative measures. The COVID-19 infection is often treated with current drugs such as Remdesivir, steroids, tocilizumab, favipiravir, and ivermectin. However, the immunosuppressive effects of these medicines might worsen COVID-19 symptoms and put the lives of immunocompromised individuals in peril. Thus, it is important to sustain a robust immune system when undergoing therapy for COVID-19. Herbal treatment has the potential to accomplish this objective. Objective: The current investigation involves the preparation of polyherbal syrup containing various medicinal plants such as ephedra, diascorea, ginger, echinacea, garlic, rhubarb, and glycyrrhiza for the effective control of the COVID-19 infection. Methods: All varieties of the individual plant powders (200 g) were treated to a 7 day maceration in aqueous ethanol (70:30) in a percolator at room temperature with intermittent vigorous shaking at room temperature and storage of the extract in a dark room. The mixture was run through a muslin cloth and then a Whatman qualitative grade 1 filter paper to produce the filtrate. The filtrate was evaporated to a thick paste-like consistency at 370 °C under decreased pressure in a rota evaporator connected to a vacuum pump. After that, each individual extract was collected and kept in airtight jars at 4°C. According to the Indian Pharmacopoeia, simple syrup (66.67% w/v) of polyherbal extract was prepared. The oral administration of polyherbal syrup was carried out at varied doses of 0.5 ml, 1 ml, and 1.5 ml to infected golden Syrian hamsters from the 7th day for one week after infection reached its peak. Results: When compared to the infection control group, the results revealed that the viral load was significantly reduced by 79.1% when treated with polyherbal syrup. A histological examination of the infected hamster lung on days 7, 10, and 13 demonstrated that polyherbal syrup significantly decreased viral load in a dose-dependent manner. Conclusion: It is inferred that the polyherbal syrup formulation demonstrates efficacy in the prevention of COVID-19 infection during its first stages and may serve as a potential contender for SARSCoV- 2 due to its immunomodulatory properties.

Safety and efficacy of a phage cocktail on murine wound infections caused by carbapenem-resistant Klebsiella pneumoniae

ObjectivesCarbapenem-resistant Klebsiella pneumoniae (CRKP) is a leading pathogen causing difficult-to-treat, healthcare-associated wound infections. Phages are an alternative approach against CRKP. This study established murine wound infection models with a CRKP clinical strain of sequence type 11 and capsular type KL64, which is the dominant type in China, carrying genes encoding KPC-2 and NDM-1 carbapenemases. MethodsA cocktail was made comprising three lytic phages of different viral families against the strain. The phage cocktail restricted bacterial growth for 10 hours in vitro. The efficacy and safety of the phage cocktail in treating a murine wound CRKP infection were then evaluated. Mice were randomly assigned into four groups (16 for each) comprising a phage treatment group, infected with bacteria and 30 minutes later with phages, and three control groups administered with PBS (negative control), bacteria (infection control), or phages (phage control) on the wound. Wound tissues were processed for counting bacterial loads on days 1, 3, and 7 post-infection and examined for histopathological change on days 3 and 7. Two remaining mice in each group were monitored for wound healing until day 14. ResultsCompared with the infection control group, the wound bacterial load in the phage treatment group decreased by 4.95 × 102 CFU/g (> 100-fold; P < 0.05) at day 7 post-treatment, and wounds healed on day 10, as opposed to day 14 in the infection control group. No adverse events associated with phages were observed. ConclusionThe phage cocktail significantly reduced the wound bacterial load and promoted wound healing with good safety.

Effects of dietary water-soluble extract of rosemary supplementation on growth performance and intestinal health of broilers infected with Eimeria tenella.

This study was conducted to explore the effect of dietary supplementation of water-soluble extract of rosemary (WER) on growth performance and intestinal health of broilers infected with Eimeria tenella (E. tenella), and evaluate the anticoccidial activity of WER. 360 1-d-old Chinese indigenous male yellow-feathered broiler chickens were randomly allocated to six groups: blank control (BC) group and infected control (IC) group received a basal diet; positive control (PC) group, received a basal diet supplemented with 200mg/kg diclazuril; WER100, WER200, and WER300 groups received a basal diet containing 100, 200, and 300mg/kg WER, respectively. On day 21, all birds in the infected groups (IC, PC, WER100, WER200, and WER300) were orally gavaged with 1mL phosphate-buffered saline (PBS) of 8 × 104 sporulated oocysts of E. tenella, and birds in the BC group were administrated an aliquot of PBS dilution. The results showed that dietary supplementation of 200mg/kg WER increased the average daily gain of broilers compared to the IC group from days 22 to 29 (P < 0.001). The anticoccidial index values of 100, 200, and 300mg/kg WER were 137.49, 157.41, and 144.22, respectively, which indicated that WER exhibited moderate anticoccidial activity. Compared to the IC group, the groups supplemented with WER (100, 200, and 300mg/kg) significantly lowered fecal oocyst output (P < 0.001) and cecal coccidia oocysts, alleviated intestinal damage and maintained the integrity of intestinal epithelium. Dietary supplementation with WER significantly improved antioxidant capacity, elevated the levels of secretory immunoglobulin A, and diminished inflammation within the cecum, particularly at a dosage of 200mg/kg. The results of this study indicated that dietary supplementation with 200mg/kg WER could improve broiler growth performance and alleviate intestinal damage caused by coccidiosis.

Ovomucin and its hydrolysates differentially influenced colitis severity in Citrobacter rodentium-infected mice.

Egg white protein ovomucin and its hydrolysates were previously reported to exhibit anti-inflammatory and anti-adhesive activities. However, their potential to regulate pathogen colonization and disease severity has not been fully characterized. To investigate the effects of ovomucin (OVM) and its hydrolysates including ovomucin-Protex 26L (OP) and -pepsin/pancreatin (OPP) on host resistance to pathogen infection, a well-documented colitis model in mice for attaching and effacing E. coli pathogens, Citrobacter rodentium, was used in the current study. C57Bl/6J female mice were fed on a basal diet supplemented with OVM or its hydrolysates for 3 weeks prior to the C. rodentium challenge, with the dietary treatments continued for seven days. Body weight was not affected throughout the experimental period. OP supplementation resulted in lower (P < 0.05) pathogen loads at 7 dpi. Attenuated colitis severity was observed in mice that received OVM and OP, as indicated by reduced colonic pathological scores and pro-inflammatory responses compared with the infected control group. In contrast, OPP consumption resulted in enhanced C. rodentium colonization and disease severity. Notably, reduced microbial diversity indices of the gut microbiota were observed in the OPP-supplemented mice compared with the OVM- and OP-supplemented groups. This study showed the potential of OVM and OP to alleviate the severity of colitis induced by infection while also suggesting the opposite outcome of OPP in mitigating enteric infection.

Endocervical miRNA-142 expression in patients with pelvic inflammatory disease positive for Chlamydia trachomatis

Abstract Background: The most common cause of sexually transmitted diseases that can lead to pelvic inflammatory disease (PID) and infertility is Chlamydia trachomatis. Chlamydial infections are characterized by multifocality and polymorphism changes. Chlamydia causes inflammation in the adult urethra and cervix with the possibility of serious complications, and can cause perinatal infections in infants. Highly conserved single-stranded noncoding RNAs called micro-RNAs (miRNAs), which control gene expression, have now been identified as significant effect in disease conditions. MiRNAs play important regulatory roles in various biological processes for example, differentiation, development, proliferation, apoptosis, cell cycle control, and metabolism. Changes in miRNA expression have been identified in many diseases, such as cardiac and autoimmune disorders, schizophrenia, and cancer. In this cross-sectional pilot investigation, samples were taken from control women who were not infected and from symptomatic women who were infected with C. trachomatis. Objectives: This work investigates the changes in the host miRNA-142 expression profile during chlamydial infection and not infected control group women Materials and Methods: The study included 200 female patients who were sampled, including endocervical swab from each female patient. Patients ranged in age from 20 to 45 years. A control group of 25 healthy ladies was also used. Chlamydia trachomatis was determined by conventional polymerase chain reaction (PCR) whereas miRNA-142 was detected by RT-qPCR. Results: All 200 collected endocervical swab specimens were diagnosed by the gynecologists as having PID. From all 200 collected endocervical swab specimens, it was found that only 22/200 (11%) were positive for C. trachomatis and the current study shows that the expression of the micro-142 gene elevated in C. trachomatis patients when compared to the control group, with the expression of the gene increasing more than 40% when compared to the control group. Conclusions: This study emphasizes the relevance of C. trachomatis infection in the female from 20 to 45 years in population of Iraq in Hilla city between February and August 2022. It also demonstrates the diversity of miRNA-142 expression involved in genital infection.

Respiratory infection risk in primary Sjögren's syndrome complicated with interstitial lung disease: a retrospective study.

To explore clinical and laboratory characteristics of primary Sjögren's syndrome (pSS) complicated with interstitial lung disease (ILD) and investigate the risk factors for respiratory infections in pSS-ILD. A cohort of 162 pSS-ILD patients in Peking University People's Hospital from 2015 to 2020 were included, and all medical records were completely collected. We screened 53 patients suffering from respiratory infections as study cases, compared with 109 age- and sex-matched controls. Differences between infection group and control group were compared. Univariate and multivariate binary logistic regression tests were conducted to identify potential risk factors for respiratory infections in pSS-ILD patients. Among 162 pSS-ILD patients, 32.72% (53/162) suffered from respiratory infections. The most frequent type of ILD was nonspecific interstitial pneumonia (32.08%, 51/159), and the most common type of pathogen was bacteria (64.25%, 34/53). Infection group showed higher levels of ESSDAI (P < 0.001), CRP (P < 0.001), ESR (P = 0.003), and C3 (P = 0.020) but lower level of DLCO-SB (P = 0.015). Univariate logistic model revealed that PAH and the use of glucocorticoid increased infection risk in pSS-ILD patients. On multivariate logistic regression analysis, PAH (OR = 3.993, 95% CI = 1.192-13.373, P = 0.025) and severe reduction of DLCO (DLCO-SB < 40%, OR = 4.625, 95% CI = 1.281-16.702, P = 0.019) were significantly associated with increased risk of respiratory infections in pSS-ILD patients. Among pSS-ILD patients, the most frequent type of ILD was nonspecific interstitial pneumonia. In patients with infection, bacteria were the most common pathogen. Higher levels of ESSDAI, CRP, ESR, and C3 may be correlated with increased infection risk. PAH and reduction of DLCO were identifiedas independent risk factors. Key Points • ILD and infectious diseases severely affect pSS patient conditions. • Higher levels of ESSDAI, CRP, ESR, and C3 may be correlated with increased infection risks in pSS-ILD. • PAH and reduction of DLCO were identified as independent risk factors for lower respiratory infection.

Resveratrol protects against Schistosoma mansoni-induced liver fibrosis by targeting the Sirt-1/NF-κB axis

Hepatic schistosomiasis is a prevalent form of chronic liver disease that drastically affects human health. Nevertheless, an antifibrotic drug that could suppress the development of hepatic fibrosis does not exist yet. The current study aimed to evaluate the effect of resveratrol, a natural polyphenol with multiple biological activities, on Schistosoma mansoni (S. mansoni)-induced hepatic fibrosis and delineate the underlying molecular mechanism. Swiss male albino mice were randomly assigned into infected and non-infected groups. Hepatic schistosomiasis infection was induced via exposure to S. mansoni cercariae. 6 weeks later, resveratrol was administrated either as 20 mg/kg/day or 100 mg/kg/day for 4 weeks to two infected groups. Another group received vehicle and served as infected control group. At the end of the study, portal hemodynamic, biochemical, and histopathological evaluation of liver tissues were conducted. Remarkably, resveratrol significantly reduced portal pressure, portal and mesenteric flow in a dose-dependent manner. It improved several key features of hepatic injury as evidenced biochemically by a significant reduction of bilirubin and liver enzymes, and histologically by amelioration of the granulomatous and inflammatory reactions. In line, resveratrol reduced the expression of pro-inflammatory markers; TNF-α, IL-1β and MCP-1 mRNA, together with fibrotic markers; collagen-1, TGF-β1 and α-SMA. Moreover, resveratrol restored SIRT1/NF-κB balance in hepatic tissues which is the main switch-off control for all the fibrotic and inflammatory mechanisms. Taken together, it can be inferred that resveratrol possesses a possible anti-fibrotic effect that can halt the progression of hepatic schistosomiasis via targeting SIRT1/ NF-κB signaling.

FNIRS-based evaluation of the impact of SARS-CoV-2 infection central auditory processing.

Coronavirus disease-2019 due to SARS-CoV-2 infection has been associated with neurological and neuropsychiatric illnesses as well as auditory system problems. In this study, we aimed to evaluate the impact of SARS-CoV-2 infection on the central auditory system by assessing the hemodynamic activation changes using functional near-infrared spectroscopy (fNIRS). Three participants who had SARS-CoV-2 infection (study group) and four participants who had no SARS-CoV-2 infection (control group) were included in the study. During the auditory oddball task in which two different frequencies of tonal stimulation were presented at 80dB HL, the participants were asked to pay attention to the rare tonal stimulation and mentally count these target stimuli throughout the task. During this task, oxygenated hemodynamic response functions were evaluated with fNIRS. Significantly increased oxygenated hemodynamic responses were observed in both groups during the task (p<.05), which was significantly higher in the study group (p<.05). Significantly more HbO activation was observed in the vmPFC, superior temporal gyrus, and medial temporal gyrus in the study group compared to controls (p<.05). Significantly higher hemodynamic activation was observed in the right hemisphere in both groups, which was significantly higher in the study group (p<.05). SARS-CoV-2 infections may impact on central auditory processing or auditory attention due to changes in oxyhemoglobin levels in the frontal and temporal brain regions. It seems that SARS-CoV-2 infection is associated with an additional load on neural activity, and difficulties in focusing in auditory attention, following speech and hearing in noise as well as increased effort to perceive auditory cues.

Probiotic Enterococcus faecalis surface-delivering key domain of EtMIC3 proteins: immunoprotective efficacies against Eimeria tenella infection in chickens.

Enterococcus faecalis MDXEF-1 strain was used as host bacteria to deliver surface-displayed key domains BC1 and C4D of the Eimeria tenella microneme 3 (EtMIC3) protein, BC1 fusion to dendritic cell (DC) targeting peptides (DCpep) (DC-BC1) and DC-C4D, respectively. The expression of target proteins in MDXEF-1/BC1-CWA, MDXEF-1/DC-BC1-CWA, MDXEF-1/C4D-CWA, and MDXEF-1/DC-C4D-CWA was verified. The systemic immune responses induced by oral immunization with recombinant E. faecalis were recorded. The immunized chickens were challenged with 4.0 × 104 E. tenella-sporulated oocysts. The immune protections were evaluated by calculating average weight gain and oocyst reduction rate, observing gross and histopathological changes in the cecal tissues, and quantifying levels of inflammatory cytokines in the cecal tissues. The results showed that proteins BC1, C4D, DC-BC1, and DC-C4D were expressed on the surface of E. faecalis, respectively. Oral immunizations with recombinant E. faecalis, especially MDXEF-1/DC-BC1-CWA and MDXEF-1/DC-C4D-CWA, stimulated higher levels of antigen-specific humoral and intestinal mucosal immune responses, higher levels of Th1, Th2, and Th17-type cytokines, and higher proliferation of peripheral blood lymphocytes than phosphate-buffered saline (PBS) and vector control groups (P < 0.01). The groups immunized with four recombinant E. faecalis showed better protective effects against homologous infection than the vector control and infection control groups, and the MDXEF-1/DC-BC1-CWA group displayed the best effects. These results demonstrated that probiotic E. faecalis delivering DCpep fused with the key domain of the EtMIC3 protein could be a potential approach for the prevention of Eimeria infection. IMPORTANCE Avian coccidiosis caused by Eimeria brings huge economic losses to the poultry industry. Although live vaccines and anti-coccidial drugs were used for a long time, Eimeria infection in chicken farms all over the world commonly occurred. The exploration of novel, effective vaccines has become a research hotspot. Eimeria parasites have complex life cycles, and effective antigens are particularly critical to developing anti-coccidial vaccines. Microneme proteins (MICs), secreted from microneme organelles located at the parasite apex, are considered immunodominant antigens. Eimeria tenella microneme 3 (EtMIC3) contains four conserved repeats (MARc1, MARc2, MARc3, and MARc4) and three divergent repeats (MARa, MARb, and MARd), which play a vital role during the Eimeria invasion. Enterococcus faecalis is a native probiotic in animal intestines and can regulate intestinal flora. In this study, BC1 and C4D domains of EtMIC3, BC1 or C4D fusing to dendritic cells targeting peptides, were surface-displyed by E. faecalis, respectively. Oral immunizations were performed to investigate immune protective effects against Eimeria infection.

Therapeutic efficacy of β-sitosterol treatment on Trypanosoma congolense infection, anemia development, and trans-sialidase (TconTS1) gene expression.

African animal trypanosomiasis hinders sustainable livestock productivity in sub-Saharan Africa. About 17 million infected cattle are treated with trypanocides annually but most of the drugs are associated with drawbacks, necessitating the search for a promising chemotherapeutic agent. In this study, the effects of β-sitosterol on Trypanosoma congolense infection were investigated along with its effect on the trans-sialidase gene expressions. Oral treatment with β-sitosterol at 15 and 30 mg/kg body weight (BW) for 14 days significantly (p < 0.05) reduced parasitemia and ameliorated the parasite-induced anemia. Also, the parasite-induced increase in serum urea level and renal histopathological damage scores in addition to renal hypertrophy was significantly (p < 0.05) reverted following treatment with 30 mg/kg BW β-sitosterol. The compound also significantly (p < 0.05) down-regulated the expression of TconTS1 but not TconTS2, TconTS3, and TconTS4. Correlation analysis between free serum sialic acid with the TconTS1 and TconTS2 gene variants revealed negative correlations in the β-sitosterol-treated groups although they were non-significant (p > 0.05) in the group treated with 15 mg/kg BW β-sitosterol. Similarly, a non-significant negative (p > 0.05) correlation between the biomolecule and the TconTS3 and TconTS4 gene variants was observed in the β-sitosterol-treated groups while positive correlations were observed in the infected untreated control group. The observed effect of β-sitosterol on T. congolense infection could make the compound a possible template for the design of novel trypanocides.

B-007 Measurement of Cytokines in Bronchoalveolar Lavage (BAL) Fluid and CSF Supports Model Development for Discovery Programs

Abstract In our laboratory, increased demand for cytokine measurements to support both safety and efficacy endpoints across discovery programs has also resulted in cytokine analyses in biological fluids other than routine serum or plasma samples. Recently we have evaluated cytokines in bronchoalveolar lavage (BAL) fluid and CSF to support a mouse respiratory infection and a rat neuroinflammatory model, respectively. Fit-for-purpose validation of these sample types is necessary to confirm cytokine methods are suitable for use, including selection of optimal sample dilutions to reduce or eliminate any matrix effects or interferences. Meso Scale Discovery® (MSD Gaithersburg, MD) methodology was used for measuring cytokines and validation parameters included precision and accuracy of calibration curves (CV% and %Relative Error &amp;lt;20%) to confirm LLOQ and ULOQ for each cytokine method, precision of results for quality control and study samples (CV ≤ 20%) and linearity of response in samples. In development of the mouse respiratory infection model, cytokines IL-1β, IL-6, KC/GRO and MCP-1 were increased in BAL fluid (up to 2892-fold) 20 h post-infection compared to sham and vehicle control groups. Once the model was established, treatment of mice with Compound A demonstrated statistical decreases in these cytokines in BAL fluid compared to an infected control group. Dilutional linearity demonstrated that a minimal required dilution of 1:2 was required for mouse BAL fluids. In support of development of the rat neuroinflammatory model, cytokines IL-1β, IL-6, KC/GRO, MCP-1 and TNF-α were increased (325X, 13X, 28X, 23X and 7.7X, respectively) in CSF compared to plasma at 24 h post treatment. Dilutional linearity evaluations demonstrated that CSF samples can be run neat but a 1:4 dilution was required for plasma samples. In conclusion, fit-for-purpose validation confirmed MSD methods were acceptable for use in supporting these drug discovery models.

Standardized approach to CPAP therapy in the delivery room in full-term infants with congenital infection: observational research

Background. At the present day, there are no protocols and guidelines on the therapeutic approaches of respiratory disorders in full-term infants in the delivery room. This concerns indications for the choice of respiratory therapy, parameters and its duration for deciding on effectiveness.The aim of the study is — to evaluate the effectiveness and safety of the standardized CPAP (continuous positive airway pressure) protocol therapy of transient tachypnea of newborns (TTN) in the delivery room in full-term infants with congenital infection.Methods. An observational clinical study of the efficacy and safety of the standardized CPAP therapy protocol developed by us for children with TTN in the delivery room in children with congenital infection, used during 6 months of 2021 in comparison with the historical control group recruited according to the same criteria for 12 months of 2020. The assessment of cerebral damage, the nature and duration of respiratory therapy and the duration of hospitalization of patients in comparison with those in the historical control group was carried out.Results. In 2020, out of 3901 full-term infants, CPAP was performed in the delivery room for 223 children, 31 of them with infection (control group). In 2021, out of 2057 full-term CPAP was performed on 140 children, 15 of them with infection (study group). A comparative analysis of the groups revealed no statistical differences in anthropometric data, gestational age, gender, APGAR score, as well as differences in the history of pregnancy and childbirth. The analysis of clinical characteristics also showed no differences between the groups (control group versus study group) in the frequency of cerebral ischemia (25 (80.6%) of the control group versus 9 (60.0%) of the study group; p = 0.135) and subependymal hemorrhage (6 (19.3%) of the control group versus 3 (20.0%) of the study group; p = 0.959), the frequency of ventilation (10 (32.2%) of the control group versus 5 (33.3%) of the study group; p = 0.942) and its duration (1 (1; 1.75) of the control group versus 2 (1.25; 2) days of the study group; p = 0.211), the frequency of BinCPAP (9 (29.3%) of the control group versus 5 (33.3%) of the study group; p = 0.434) and its duration (1 (1; 2) of the control group versus 1 (1; 1) day of the study group; p = 0.434) and the total duration of hospitalization (14 (10; 16) control groups versus 12 (12; 14) days of the study group; p = 0.429).Conclusion. The conducted study does not exclude that the standardized protocol of CPAP therapy in the delivery room for congenital infection is ineffective and does not affect the severity and duration of the disease, however, in combination with the data of the same protocol for TTN described in our previous studies, the high frequency of TTN and the low frequency of infection, the impossibility of their early differentiation allows assume, that the presented method can be considered as a universal respiratory strategy for full-term newborns with respiratory insufficiency in the delivery room.

Attenuation of pathogenesis of Eimeria stiedae sporulated oocysts using Egyptian alginate propolis nanoparticles

BackgroundCoccidiosis is a costly and widespread infectious disease that affects mammals and causes huge losses for the global rabbit meat industry. This study evaluated the potency of Egyptian alginate propolis nanoparticles (NPs) in attenuating the infectivity of Eimeria stiedae sporulated oocysts. The gelification method was used to prepare alginate propolis NPs, which were then characterized using a transmission electron microscope and zeta potential analysis.ResultsThe results revealed that the zeta potential of the prepared alginate propolis NPs increased from − 60.60 ± 9.10 mV to –72.26 ± 6.04 mV. The sporulated oocysts were treated with 50 mg/mL of the alginate propolis NPs. Thereafter, the treated oocysts were tested for their ability to infect rabbits. The rabbits were divided into three groups: the healthy control (G1) group, the infected control (G2) group, and the treated oocyst-infected (G3) group. The rabbits were sacrificed 43 days post-infection (dpi). The infectivity of the oocysts was assessed. The treated oocyst-infected rabbits exhibited slight abdominal distension and dullness symptoms. The G3 group had no oocyst output, with a 100% reduction from 41 dpi until the end of the experiment. Immunologically, the IgG level of the G2 group gradually increased (p ≤ 0.05) much more than that of the G3 group. The IL-12 level in the G3 group significantly increased from 16 dpi until the end of the experiment, nearly reaching the level in healthy animals. Decreased CD4+ and CD8+ immunolabelling was observed in the liver sections of the group infected with the alginate propolis NP-treated oocysts, and there was a remarkable improvement in the histopathological parameters.ConclusionsThese data indicate that Alg propolis NPs are sufficient to reduce the infectivity of E. stiedae oocysts.

Temporal changes in fecal microbiota of patients infected with COVID-19: a longitudinal cohort

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a multifaceted disease potentially responsible for various clinical manifestations including gastro-intestinal symptoms. Several evidences suggest that the intestine is a critical site of immune cell development, gut microbiota could therefore play a key role in lung immune response. We designed a monocentric longitudinal observational study to describe the gut microbiota profile in COVID-19 patients and compare it to a pre-existing cohort of ventilated non-COVID-19 patients.MethodsFrom March to December 2020, we included patients admitted for COVID-19 in medicine (43 not ventilated) or intensive care unit (ICU) (14 ventilated) with a positive SARS-CoV-2 RT-PCR assay in a respiratory tract sample. 16S metagenomics was performed on rectal swabs from these 57 COVID-19 patients, 35 with one and 22 with multiple stool collections. Nineteen non-COVID-19 ICU controls were also enrolled, among which 14 developed ventilator-associated pneumonia (pneumonia group) and five remained without infection (control group). SARS-CoV-2 viral loads in fecal samples were measured by qPCR.ResultsAlthough similar at inclusion, Shannon alpha diversity appeared significantly lower in COVID-19 and pneumonia groups than in the control group at day 7. Furthermore, the microbiota composition became distinct between COVID-19 and non-COVID-19 groups. The fecal microbiota of COVID-19 patients was characterized by increased Bacteroides and the pneumonia group by Prevotella. In a distance-based redundancy analysis, only COVID-19 presented significant effects on the microbiota composition. Moreover, patients in ICU harbored increased Campylobacter and decreased butyrate-producing bacteria, such as Lachnospiraceae, Roseburia and Faecalibacterium as compared to patients in medicine. Both the stay in ICU and patient were significant factors affecting the microbiota composition. SARS-CoV-2 viral loads were higher in ICU than in non-ICU patients.ConclusionsOverall, we identified distinct characteristics of the gut microbiota in COVID-19 patients compared to control groups. COVID-19 patients were primarily characterized by increased Bacteroides and decreased Prevotella. Moreover, disease severity showed a negative correlation with butyrate-producing bacteria. These features could offer valuable insights into potential targets for modulating the host response through the microbiota and contribute to a better understanding of the disease's pathophysiology.Trial registrationCER-VD 2020–00755 (05.05.2020) & 2017–01820 (08.06.2018).

Characteristics of peripheral lymphocyte subsets in patients with different stages of schistosomiasis japonica.

Immune cells are important for the development of schistosomiasis japonica and are also critical for the treatment of schistosomiasis. The immune cells in the peripheral blood help assess the immune state. The peripheral lymphocytes in schistosomiasis mansoni were well studied; however, immune cells in patients with different stages of schistosomiasis japonica are not well analysed. Here, we performed a preliminary study to explore characteristics of peripheral lymphocyte subsets in patients with different stages of schistosomiasis japonica. 135 patients with Schistosoma japonicum infection and 25 healthy volunteers were included in this study, including 84 patients with chronic S. japonicum infection and 51 patients with advanced S. japonicum infection. Flow cytometry analysis was performed to evaluate peripheral lymphocytes including T cells, B cells, and natural killer (NK) cells. Blood routine and liver function test data were analysed. Ultrasound examination was used to access liver fibrosis according to the World Health Organization standard about ultrasound in schistosomiasis. Demographic data analysis suggested there was no difference in age and gender in patients with S. japonicum infection and health control group. Liver function tests showed that patients with advanced schistosomiasis had a higher incidence of liver function abnormality and blood lipid than those with chronic schistosomiasis. Blood routine results reflected that haemoglobin, red blood cells, platelets, as well as lymphocytes in the advanced group were significantly less than that in the chronic group. Furthermore, flow cytometry analysis indicated that the percentage of CD4+ T cells was lower in the advanced group, but the percentage of CD19+ B cells was higher in the advanced group. In addition, the number of CD3+ T cells, CD3+ CD4+ T cells, CD3+ CD8+ T cells, and NK cells was less in the advanced group when compared with those in the chronic group. In addition, there was a correlation between the decrease in CD4+ T cells and more severe fibrosis on ultrasound images. Our results indicated that the immune state in the peripheral is different in different stages of S. japonicum infection. Lymphocyte subset analysis has potential to facilitate differential diagnosis of different stages of schistosomiasis japonica and even to be a prognostic factor.

Ameliorative synergistic therapeutic effect of gallic acid and albendazole against Trichinella spiralis muscular phase infection and assessment of their effects on hepatic and cardiac tissues in male mice

Trichinellosis is a serious food-borne parasitic disease with serious community health effects, mainly causing muscle damage with no recent approved treatment. This study aimed to assess the therapeutic effect of gallic acid (GA) as a potent antioxidant against the encysted phase of Trichinella spiralis in male (BALB/c) mice alone or combined with albendazole (ALB) and to detect their synergistic effects on the histology and ultrastructure of skeletal and cardiac muscles and some biochemical blood analyses. Forty male mice were randomly divided into five groups (8 mice/group). 1st group: the negative control received only distilled water, 2nd group: the positive control (infected control group without treatment), 3rd group: infected group plus treatment with ALB (50 mg Kg−1 orally), and 4th group: infected group and then treated with GA (30 mg Kg−1 orally) and finally 5th infected group treated with a combination of both ALB and GA. Aspartate and Alanine aminotransferase, Lactate dehydrogenase, alkaline phosphatase, C-reactive protein, Interleukin-4 and Creatine kinase were used as biochemical markers of hepatic and cardiac toxicity and inflammation. Malondialdehyde level, catalase, superoxide dismutase, and glutathione peroxidase were evaluated in heart tissue homogenates beside histological and ultra-structural examination of heart and skeletal muscles beside parasitological analyses. Results showed that the reduction % of Trichinella sp. larvae g−1 in muscles of the group treated with the combination of GA and ALB showed overall reduction percentages. Oral administration of 30 mg kg1 of GA led to infection reduction of T. spiralis than ALB treated group. Both administration of ALB beside GA showed the best treatment group that resulted in high infection reduction besides amelioration of both biochemical markers and restoration of histological and ultrastructures to normal state. In conclusion, GA is highly effective against T. spiralis which could be a promising alternative antioxidant drug and the GA effect was higher in the case of combination with ALB. This experiment provides a basis for further exploration of potent activities of other antioxidants against different phases of T. spiralis and the reduction of any health hazards prospectively.

Transcriptome and the gut microbiome analysis of the impacts of Brucella abortus oral infection in BALB/c mice

Brucellosis is a zoonotic infectious disease caused by Brucella spp, which could cause serious economic losses to animal husbandry and threaten human public health. Ingestion of contaminated animal products is a common way to acquire Brucella infection in humans, while research on effect of oral Brucella infection on host gut microbiota and the gene expression in intestinal tissues is limited. In the present study, 16S rRNA sequencing and RNA sequencing were conducted to explore gut microbiota and expression profiles of mRNAs in the colon of BALB/c mice, which were infected by Brucella abortus 2308. The fecal samples were collected at 7 and 28 days post infection to observe changes in the gut microbiota during Brucella infection. In the alpha diversity analysis, significantly increased Chao 1 index was observed at 28 days after Brucella infection. The Bray–Curtis distancebased principal coordinate analysis indicated that the WT group showed a separation from the Brucella infection groups. In addition, analysis of composition of microbes revealed that Prevotellaceae_NK3B31_group were more abundant in 1 week and 4 week infection groups, while Turicibacter was only more abundant in 4 week infection group. Based on the RNA-seq assay, a total of 45 differentially expressed genes were detected between Brucella abortus infection group and control group. Furthermore, KEGG pathway enrichment analysis showed that protein processing in endoplasmic reticulum, Legionellosis, Spliceosome, Hippo signaling pathway and Influenza A were significantly enriched in response to Brucella abortus infection. Our finding will help to improve the knowledge of the mechanisms underlying Brucella infection and may provide novel targets for future treatment of this pathogen infection.

Dietary Olive Leaf Extract Differentially Modulates Antioxidant Defense of Normal and Aeromonas hydrophila-Infected Common Carp (Cyprinus carpio) via Keap1/Nrf2 Pathway Signaling: A Phytochemical and Biological Link.

Olive leaves are an immense source of antioxidant and antimicrobial bioactive constituents. This study investigated the effects of dietary incorporation of olive leaf extract (OLE) on the growth performance, hematobiochemical parameters, immune response, antioxidant defense, histopathological changes, and some growth- and immune-related genes in the common carp (Cyprinus carpio). A total of 180 fish were allocated into four groups with triplicate each. The control group received the basal diet without OLE, while the other three groups were fed a basal diet with the OLE at 0.1, 0.2, and 0.3%, respectively. The feeding study lasted for 8 weeks, then fish were challenged with Aeromonas hydrophila. The results revealed that the group supplied with the 0.1% OLE significantly exhibited a higher final body weight (FBW), weight gain (WG%), and specific growth rate (SGR) with a decreased feed conversion ratio (FCR) compared to the other groups (p < 0.05). An increase in immune response was also observed in the fish from this group, with higher lysosome activity, immunoglobulin (IgM), and respiratory burst than nonsupplemented fish, both before and after the A. hydrophila challenge (p < 0.05). Similarly, the supplementation of the 0.1% OLE also promoted the C. carpio's digestive capacity pre- and post-challenge, presenting the highest activity of protease and alkaline phosphatase (p < 0.05). In addition, this dose of the OLE enhanced fish antioxidant capacity through an increase in the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) and decreased hepatic lipid peroxidation end products (malondialdehyde-MDA), when compared to the control group, both pre- and post-infection (p < 0.05). Concomitantly with the superior immune response and antioxidant capacity, the fish fed the 0.1% OLE revealed the highest survival rate after the challenge with A. hydrophila (p < 0.05). A significant remarkable upregulation of the hepatic sod, nrf2, and protein kinase C transcription levels was detected as a vital approach for the prevention of both oxidative stress and inflammation compared to the infected unsupplied control group (p < 0.05). Interestingly, HPLC and UPLC-ESI-MS/MS analyses recognized that oleuropein is the main constituent (20.4%) with other 45 compounds in addition to tentative identification of two new compounds, namely oleuroside-10-carboxylic acid (I) and demethyl oleuroside-10-carboxylic acid (II). These constituents may be responsible for the OLE exerted potential effects. To conclude, the OLE at a dose range of 0.66-0.83 g/kg w/w can be included in the C. carpio diet to improve the growth, antioxidant capacity, and immune response under normal health conditions along with regulating the infection-associated pro-inflammatory gene expressions, thus enhancing resistance against A. hydrophila.

Role of Programmed Cell Death-1 and Programmed Cell Death Ligand-1 immune checkpoint biomarkers among chronic Hepatitis C virus patients under Hemodialysis

Background: Hepatitis C virus (HCV) infection is one of the most common infections associated with chronic kidney disease (CKD) patients undergoing hemodialysis (HD) in Iraq. Aim of the study: To determine the prognostic factor value of Programmed Cell Death-1 (PD-1) and Programmed Cell Death Ligand-1 (PD-L1) immune checkpoint biomarkers among CKD patients with HCV infection under HD. Methodology: ELISA technique was used for the measurement of the above-mentioned biomarkers in the serum of 90 Iraqi patients. The participants were divided into three groups; Group I included 30 patients infected with HCV without antiviral treatment, group II included 30 patients infected with HCV with recent/previous antiviral treatment, and Group III included 30 patients without viral infection (control group). Results: Serum levels of the measured biomarkers were elevated among all the participants, and highly statistically significant differences were found between patients with no treatment. The area under the curve (AUC) of PD-1 was 99% and for PD-L1 was 96%. Conclusions: The PD-1 and PD-L1 immune checkpoint biomarkers have excellent prognostic factor value as predictors for patients with CKD on HD infected with HCV.

The effects of intracellular iron availability on the outcome of Toxoplasma gondii infection in mice.

Toxoplasma gondii (T. gondii) is a parasite that obtains the iron it needs for its own metabolism from the host-cell iron pool. In this work, we aimed to investigate if iron supplementation or deficiency affected the course of T. gondii infection. Eighty mice were divided into four groups, each with 20 animals: Group (I): Uninfected control group. Group (II): Infected control group: injected with Phosphate buffered saline. Group (III): Infected group: received iron sucrose treatment. Group (IV): Infected group: treated with deferoxamine. Quantitative PCR studies were performed on days 3 and 8 post-infection to detect the expression of iron metabolism genes (hamp and ferroprotin) and immune-histochemical analysis to study the percentage of TNF-α and TGF-β tissue expression. Iron supplementation induced progressions of infection evident by increased tissue expression of pro-inflammatory cytokine TNF-α and downregulation of TGF-β which is mostly linked to suppression of the inflammatory process caused by T. gondii. Increased expression of TGF-β and decreased expression of TNF-α was noticed when iron deprivation occurred. On day 3, we noticed increased expression in the hamp gene with iron supplementation while it decreases when the iron supply is low. On the contrary, iron deficiency increased ferroprotin gene expression whereas supplementing decreased it. On day 8, the level of expression of these genes returned to normal levels. These observations document the potential role of iron in controlling toxoplasmosis infection and indicate that the transcription of hamp and ferroprotin in T. gondii-infected cells appears to be regulated by a sophisticated indirect mechanism.