Anaplasma capra in sheep and goats on Corsica Island,France: A European lineage within A. capra clade II?

Anaplasmosis is a tick-transmitted disease due to several species of the genus Anaplasma. In 2019, we demonstrated the presence of Anaplasma capra in two deer species at a zoological park in mainland France. As we suspected its presence in Corsica, we surveyed 11 geographically distant sheep or goat farms. Using molecular tools such as nested PCR targeting 16S ribosomal RNA (rRNA), citrate synthase (gltA) and heat-shock protein (groEL) genes, we detected the presence of A. capra on 5/11 farms, in 26/108 blood samples (24%), in sheep as well as in goats. Genotyping and phylogenetic analysis of A. capra revealed that isolates from Corsica island grouped closely with A. capra isolates reported in red deer and swamp deer from a zoological reserve in mainland France, as well as in roe deer from Spain, in a separate and well supported clade within A. capra clade II. This third report of the tick-borne bacterium A. capra in Europe suggests a potentially larger presence of this pathogen on the European continent, on domestic, native as well as wild ruminants, a broad host range already described in Asian countries for this species.     

The oxidative stress response of myclobutanil and cyproconazole on Tetrahymena thermophila

Using Tetrahymena thermophila as experimental models, the oxidative stress of triazole fungicides myclobutanil (MYC) and cyproconazole (CYP) was investigated. Results showed that 24-h EC₅₀ values for MYC and CYP were 16.67 (13.37–19.65) and 20.44 (18.85–21.96) mg/L, respectively; 48-h EC₅₀ values for MYC and CYP were 14.31 (13.13–15.42) and 18.76 (17.09–20.31) mg/L, respectively. Reactive oxygen species was significantly induced and cytotoxicity was caused by MYC and CYP by increasing propidium iodide (PI) fluorescence. Damage of regular wrinkles and appearing of small holes on the cell surface were observed by SEM. Furthermore, MYC and CYP also caused notable changes in enzyme activities and mRNA levels. Overall, the present study points out that MYC and CYP lead to oxidative stress on T. thermophila. The information presented in this study will provide insights into the mechanism of triazoles-induced oxidative stress on T. thermophila.     

A novel liposome adjuvant DPC mediates Mycobacterium tuberculosis subunit vaccine well to induce cell-mediated immunity and high protective efficacy in mice

Tuberculosis (TB) is a serious disease around the world, and protein based subunit vaccine is supposed to be a kind of promising novel vaccine against it. However, there is no effective adjuvant available in clinic to activate cell-mediated immune responses which is required for TB subunit vaccine. Therefore, it is imperative to develop new adjuvant. Here we reported an adjuvant composed of dimethyl dioctadecylammonium (DDA), Poly I:C and cholesterol (DPC for short). DDA can form a kind of cationic liposome with the ability to deliver and present antigen and can induce Th1 type cell-mediated immune response. Poly I:C, a ligand of TLR3 receptor, could attenuate the pathologic reaction induced by following Mycobacterium tuberculosis challenge. Cholesterol, which could enhance rigidity of lipid bilayer, is added to DDA and Poly I:C to improve the stability of the adjuvant. The particle size and Zeta-potential of DPC were analyzed in vitro. Furthermore, DPC was mixed with a TB fusion protein ESAT6-Ag85B-MPT64(190-198)-Mtb8.4-Rv2626c (LT70) to construct a subunit vaccine. The subunit vaccine-induced immune responses and protective efficacy against M. tuberculosis H37Rv infection in C57BL/6 mice were investigated. The results showed that the DPC adjuvant with particle size of 400 nm and zeta potential of 40 mV was in good stability. LT70 in the adjuvant of DPC generated strong antigen-specific humoral and cell-mediated immunity, and induced long-term higher protective efficacy against M. tuberculosis infection (5.41 ± 0.38 log₁₀ CFU) than traditional vaccine Bacillus Calmette–Guerin (BCG) (6.01 ± 0.33 log₁₀ CFU) and PBS control (6.53 ± 0.26 log₁₀ CFU) at 30 weeks post-vaccination. In conclusion, DPC would be a promising vaccine adjuvant with the ability to stimulate Th1 type cell-mediated immunity, and could be used in TB subunit vaccine.     

Occurrence and clinical manifestations of tick-borne rickettsioses in Western Siberia: First Russian cases of Rickettsia aeschlimannii and Rickettsia slovaca infections

Rickettsia sibirica subsp. sibirica is a main agent of tick-borne rickettsioses in Western Siberia, Russia. Recently, the first cases of Rickettsia raoultii infection in patients hospitalized in Novosibirsk Province were described. The aim of this study was to establish the etiologic agents of tick-borne rickettsioses in Western Siberia during three epidemiological seasons.Clinical samples from 1008 patients hospitalized after tick bites in April–September 2017–2019 were examined by nested PCR for the presence of Rickettsia spp. All positive specimens were genetically characterized by sequencing of the gltA gene; some specimens were also genotyped based on the 16S rRNA, ompA, and ompB genes. Rickettsia spp. DNA was detected in clinical samples from 56 (5.6%) patients. Five Rickettsia species were identified: R. sibirica (n = 28), R. raoultii (n = 15), Rickettsia aeschlimannii (n = 3), “Candidatus Rickettsia tarasevichiae” (n = 2), and Rickettsia slovaca (n = 1). In addition, new unclassified Rickettsia genovariants were found in specimens from seven patients. Patients with R. raoultii infection presented rash, eschar, and high serum aminotransferase levels less frequently compared to patients with R. sibirica infections, but more frequently showed neurological symptoms. Among other patients, only persons with an R. aeschlimannii infection had rash and/or eschar, which are typical for tick-borne rickettsioses. The current study showed that R. raoultii is a common agent of tick-borne rickettsioses in Novosibirsk Province. DNA from R. aeschlimannii and R. slovaca was found in clinical samples of patients in the Russian Federation for the first time.     

Mitochondrial transfer from bone-marrow-derived mesenchymal stromal cells to chondrocytes protects against cartilage degenerative mitochondrial dysfunction in rats chondrocytes

BackgroundPrevious studies have reported that mitochondrial dysfunction participates in the pathological process of osteoarthritis (OA). However, studies that improve mitochondrial function are rare in OA. Mitochondrial transfer from mesenchymal stem cells (MSCs) to OA chondrocytes might be a cell-based therapy for the improvement of mitochondrial function to prevent cartilage degeneration. This study aimed to determine whether MSCs can donate mitochondria and protect the mitochondrial function and therefore reduce cartilage degeneration.MethodsBone-marrow-derived mesenchymal stromal cells (BM-MSCs) were harvested from the marrow cavities of femurs and tibia in young rats. OA chondrocytes were gathered from the femoral and tibial plateau in old OA model rats. BM-MSCs and OA chondrocytes were co-cultured and mitochondrial transfer from BM-MSCs to chondrocytes was identified. Chondrocytes with mitochondria transferred from BM-MSCs were selected by fluorescence-activated cell sorting. Mitochondrial function of these cells, including mitochondrial membrane potential (Δψm), the activity of mitochondrial respiratory chain (MRC) enzymes, and adenosine triphosphate (ATP) content were quantified and compared to OA chondrocytes without mitochondrial transfer. Chondrocytes proliferation, apoptosis, and secretion ability were also analyzed between the two groups.ResultsMitochondrial transfer was found from BM-MSCs to OA chondrocytes. Chondrocytes with mitochondrial from MSCs (MSCs + OA group) showed increased mitochondrial membrane potential compared with OA chondrocytes without mitochondria transfer (OA group) (1.79 ± 0.19 vs. 0.71 ± 0.12, t = 10.42, P < 0.0001). The activity of MRC enzymes, including MRC complex I, II, III, and citrate synthase was also improved (P < 0.05). The content of ATP in MSCs + OA group was significantly higher than that in OA group (161.90 ± 13.49 vs. 87.62 ± 11.07 nmol/mg, t = 8.515, P < 0.0001). Meanwhile, we observed decreased cell apoptosis (7.09% ± 0.68% vs.15.89% ± 1.30%, t = 13.39, P < 0.0001) and increased relative secretion of type II collagen (2.01 ± 0.14 vs.1.06 ± 0.11, t = 9.141, P = 0.0008) and proteoglycan protein (2.08 ± 0.20 vs. 0.97 ± 0.12, t = 8.227, P = 0.0012) in MSCs + OA group, contrasted with OA group.ConclusionsMitochondrial transfer from BM-MSCs provided protection for OA chondrocytes against mitochondrial dysfunction and degeneration through improving mitochondrial function, cell proliferation, and inhibiting apoptosis in chondrocytes. This finding may offer a new therapeutic direction for OA.     

Evaluation of a nanotechnology-based approach to induce gene-expression in human THP-1 macrophages under inflammatory conditions

Macrophages orchestrate the initiation and resolution of inflammation by producing pro- and anti-inflammatory products. An imbalance in these mediators may originate from a deficient or excessive immune response. Therefore, macrophages are valid therapeutic targets to restore homeostasis under inflammatory conditions. We hypothesize that a specific mannosylated nanoparticle effectively induces gene expression in human macrophages under inflammatory conditions without undesirable immunogenic responses. THP-1 macrophages were challenged with lipopolysaccharide (LPS, 5 μg/mL). Polyethylenimine (PEI) nanoparticles grafted with a mannose receptor ligand (Man-PEI) were used as a gene delivery method. Nanoparticle toxicity, Man-PEI cellular uptake rate and gene induction efficiency (GFP, CD14 or CD68) were studied. Potential immunogenic responses were evaluated by measuring the production of tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-6 and IL-10. Man-PEI did not produce cytotoxicity, and it was effectively up-taken by THP-1 macrophages (69%). This approach produced a significant expression of GFP (mRNA and protein), CD14 and CD68 (mRNA), and transiently and mildly reduced IL-6 and IL-10 levels in LPS-challenged macrophages. Our results indicate that Man-PEI is suitable for inducing an efficient gene overexpression in human macrophages under inflammatory conditions with limited immunogenic responses. Our promising results set the foundation to test this technology to induce functional anti-inflammatory genes.     

Normal Electrocardiogram of Bama Miniature Pigs (Sus scrofa domestica)

This study determined the normal ECG patterns and values for Bama miniature pigs. Standard limb-lead ECG were recorded from 120 clinically healthy, unanesthetized piglets (age, 2 to 4 mo). The values for the ECG parameters (mean ± 1 SD) were: heart rate, 125.56 ± 18.80 bpm; P amplitude, 0.11 ± 0.03 mV; QRS amplitude, 0.63 ± 0.31 mV; P duration, 43.99 ± 5.98 ms; QRS complex, 55.27 ± 7.02 ms; RR interval, 487.55 ± 77.32 ms; PR interval, 90.72 ± 11.94 ms; QT interval, 244.72 ± 25.27 ms; and mean electrical axis, 22.2 ± 80.3°. The P waves were predominantly positive in leads I and II and in the augmented unipolar limb aVF lead; by comparison, the QRS patterns were less uniform. The T waves were slightly positive in leads II, III, and aVF. The determination and publication of the normal ECG patterns and values of Bama minipigs facilitates understanding of the electrocardiographic changes that arise under experimental conditions.Minipigs play an essential role in biomedical and pharmaceutical research.⁸ General toxicology studies can be performed in minipigs by using the oral, cutaneous, parenteral, and inhalant routes of administration.¹ These animals are advantageous for safety pharmacology studies, particularly for studies involving the cardiovascular system, because their morphology and functional, hemodynamic, and metabolic values are similar to those of humans.^8,9,11Although more than 20 minipig breeds exist, only a few have been bred for use as standard laboratory animals.⁷ Among laboratory minipigs, Göttingen minipigs have been widely used in regulatory toxicity studies in Europe, North America, and Japan.^1,6,7 However, in China, Bama minipigs are used more frequently than are Göttingen minipigs. The Bama minipigs are native to the Guangxi province of China, has been bred as a laboratory animal, and is widely used in Chinese medical and pharmaceutical research.¹⁰During pharmaceutical research, particularly during safety pharmacology studies, serial ECG recordings are necessary components of the cardiovascular system monitoring process.^5,16 In addition, ECG recordings are well-established tools for evaluating animals with cardiac diseases.¹⁵ The reported ECG values and patterns of pigs vary somewhat,^3,4 possibly due to differences in breed, age, and body weight of the experimental animals. To our knowledge, standard ECG values have not been published for Bama minipigs, making the evaluation of study-derived ECG changes difficult. Therefore, the current study describes the normal ECG patterns and values of Bama minipigs under conventional, controlled conditions.     

慢性间断性缺氧对帕金森病模型小鼠认知功能的影响

目的：探讨慢性间断性缺氧（CIH）对帕金森病模型小鼠认知功能的影响。方法：44只雄性6周龄C57BL/6小鼠,随机分为百草枯组、CIH组、百草枯加CIH组和对照组。用缺氧-复氧循环装置模拟CIH过程,通过腹腔注射百草枯诱导制备帕金森病模型小鼠。使用Y型-电迷宫和跳台实验评价小鼠学习记忆能力;化学比色法测定小鼠海马乙酰胆碱含量和乙酰胆碱酯酶活性。结果：3个实验组的学习记忆成绩、乙酰胆碱含量和乙酰胆碱酯酶活性均明显低于对照组（P均〈0.01）,其中百草枯加CIH组明显低于百草枯组与CIH组（P均〈0.05）,百草枯组与CIH组比较差异无统计学意义（P均〉0.05）。结论：CIH加重了百草枯所致帕金森病模型小鼠认知功能下降程度,可能与海马Ach含量下降和Ach E活性降低有关。