Inflammatory factors mediated the effect of air pollution on ischemic stroke: a two-step, mediation Mendelian randomization study.

OBJECTIVE: Numerous investigations have indicated a correlation between air pollution (AP) and an elevated ischemic stroke (IS) likelihood. The existing literature does not provide a consensus about the possible link between AP and IS. A two-sample Mendelian randomization (MR) analysis was utilized to systematically measure the causal link between AP and ischemic stroke. Furthermore, the mediating impact of inflammatory factors was also performed by a two-step MR. MATERIALS AND METHODS: A two-sample MR analysis was utilized to examine the AP impact on the incidence of IS. Additionally, a two-step MR approach was carried out to account for possible mediating variables. The indirect impact was determined by employing the product approach, which included multiplying the AP impact on inflammatory factors by the inflammatory factors' impacts on IS. The MR effect was identified through inverse variance-weighted (IVW) meta-analysis of each Wald Ratio. Additionally, complementary studies were conducted using the weighted median and MR-egger approaches. RESULTS: The IVW method with random effects showed that the per unit increase in genetically predicted PM2.5 was linked to the 0.362-fold elevated ischemic stroke risk (OR: 1.362, 95% CI: 1.032-1.796, p=0.029). Furthermore, the IVM technique, incorporating random effects, demonstrated that the per unit increase in genetically predicted PM2.5 was related to an elevated Interleukin (IL)-1ß risk (OR: 1.529, 95% CI: 1.191-1.963, p=0.001), IL-6 (OR: 1.498, 95% CI: 1.094-2.052, p=0.012) and IL-17 (OR: 1.478, 95% CI: 1.021-2.139, p=0.038). IL-1ß, IL-6, and IL-17 modulated the PM2.5 impact on ischemic stroke, while the proportion mediated by them was 59.5%. CONCLUSIONS: A positive correlation between genetically predicted PM2.5 levels and elevated ischemic stroke risk is mediated by IL-1ß, IL-6, and IL-17.

Metagenomic next-generation sequencing vs. conventional detection methods for detecting the pulmonary infections.

OBJECTIVE: The absence of proper pathogen treatment in the early stages can result in missing out on treatment chances or the overuse of antibiotics, both of which are the primary factors behind fatalities caused by lung infections. In this study, we aimed to investigate the efficacy of metagenomic next-generation sequencing (mNGS) in comparison to conventional detection methods in detecting infectious pathogens. PATIENTS AND METHODS: In this retrospective study, the infection pathogens of 104 patients were examined, and 86 bronchoalveolar lavage fluid (BALF), eight pleural effusions, and ten sputum samples were collected. The conventional detection approaches and mNGS analysis were used to determine the infection pathogen profiles and their detection rates were analyzed. RESULTS: Our study showed that mNGS was more sensitive (89.42%) than the conventional detection methods (56.73%) (p < 0.001), with a 32.69% improvement in sensitivity. The efficacy of mNGS in detecting mixed infections was significantly higher than that of conventional detection methods, with a detection rate of 85.29% compared to 17.65% (p < 0.001). The study demonstrated that mNGS had a higher sensitivity than the conventional detection methods when it came to diagnosing pulmonary infections, making it a potentially useful tool for clinical diagnosis. CONCLUSIONS: Combining mNGS with other pathogenic detection techniques can be an effective way to increase the rate of detecting pulmonary infections, as well as to provide guidance for treatment adjustments. Furthermore, the timing of sample collection and antibiotic administration can influence the effectiveness of mNGS when used on BALF specimens.

Expression of Nrf2-Keap1-ARE signal pathway in traumatic lung injury and functional study.

OBJECTIVE: Traumatic lung injury (TLI) can cause inflammation and oxidative stress, or even leads to acute respiratory distress syndrome (ARDS) and death. Nuclear factor erythroid-2 related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1)-antioxidant response element (ARE) signal pathway participates in disease occurrence and progression via regulating inflammatory and oxidative stress response, but with its expression and functional roles in TLI largely unknown. MATERIALS AND METHODS: Wistar rats were randomly divided into control group, TLI group by crushing method, and Nrf2 activation group which received Nrf2 specific agonist sulforaphane 30 min before TLI treatment. Artery blood gas (ABG), wet/dry mass ratio (W/D) of lung tissues, myeloid peroxidase (MPO) and superoxide dismutase (SOD) activity of lung tissue were analyzed. Keap1 and ARE mRNA levels were tested by Real-time PCR, while Nrf2 protein was measured by Western blot. Inflammatory factors including tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) were quantified by enzyme-linked immunosorbent assay (ELISA). RESULTS: TLI model had lower ABG or SOD, higher W/D ratio, MPO value, elevated expressions of TNF-α, IL-2, and Keap1, plus decreased Nrf2 and ARE expression (p<0.05). Nrf2 activation significantly improved ABG, decreased W/D ratio and MPO value, enhanced SOD activity, decreased TNF-α and IL-2 secretion, suppressed Keap1 expression, and facilitated Nrf2 and ARE expressions (p<0.05). CONCLUSIONS: Nrf2-Keap1-ARE signal pathway can improve TLI-related pathology via modulating oxidative stress response and suppressing inflammation.

Enhanced antitumor efficacy of resveratrol-loaded nanocapsules in colon cancer cells: physicochemical and biological characterization.

OBJECTIVE: The aim of present work was to prepare resveratrol-loaded lipid-core-nanocapsule (RSV-LNC) and to characterize its ability to target the colon cancer cells. MATERIALS AND METHODS: The lipid-core nanocapsule was prepared by precipitation method. The nanoparticle was prepared and evaluated regarding physical, chemical and biological parameters. RESULTS: The average size of optimized nanocapsule was ~159 nm with a uniform size distribution index of 0.15 (PDI). The RSV-LNC showed a controlled and sustained release pattern with maximum release up to ~70% by the end of 48h study period. LNC showed an excellent cellular uptake potential. LNC showed a typical endocytosis-mediated cellular internalization process and located in the cell cytoplasm. DISCUSSION: Importantly, RSV encapsulated in a nanocapsule showed a superior anticancer effect in HT29 cancer cells than compared to that of free RSV. Consistently, RSV-LNC showed a remarkable ~36% of cell apoptosis indicating its superior anticancer effect. CONCLUSIONS: Based on the in vitro studies, RSV encapsulated in a nanocapsule showed promising potential to increase the therapeutic efficacy in colon cancer cells; however, further studies on animal models are warranted to confirm the improved effects of RSV nanoformulations.

Reduced expression of SCF in serum and follicle from patients with polycystic ovary syndrome.

OBJECTIVE: In humans, stem cell factor (SCF), produced by cumulus granulosa cells during the follicular phase, plays a crucial role in follicular development. Remarkably, polycystic ovary syndrome (PCOS), one of the main reasons affecting women fertility, is accompanied by some abnormal follicles. Is there a relationship between SCF and PCOS? This study aimed to compare the expression of SCF in follicle and serum from patients with and without PCOS undergoing in vitro fertilization (IVF) treatment and to investigate the potential relationship between aberrant SCF expression and PCOS. PATIENTS AND METHODS: Serum, follicular fluid (FF) samples and granulosa cells (GCs) from 48 patients with PCOS (PCOS group) and 62 normal ovulatory patients (control group) were collected. SCF was evaluated in FF, serum, and GCs by using enzyme-linked immunosorbent assay, immunofluorescence staining, Western blot and real-time PCR. The rates of metaphase II (MII) oocyte, fertilization, embryo cleavage and high-quality embryo between PCOS group and control group were also analyzed. RESULTS: The rates of MII oocyte and fertilization were significantly lower in PCOS group than those in control group (p < 0.05). No difference was observed for the rate of embryo cleavage and high-quality embryo in these two groups. The concentrations of SCF in serum and FF from PCOS patients were remarkably lower than those in the controls (p < 0.05). Moreover, the expressions of SCF protein and SCF mRNA in GCs from PCOS patients were also decreased compared with the controls (p < 0.05). CONCLUSIONS: PCOS patients showed a reduced SCF expression in serum and follicle, which might be associated with oocyte dysmaturity and low fertilization rate.

Different outcomes after intracytoplasmic sperm injection without oocyte activation in two patients with different types of globozoospermia.

Different outcomes after intracytoplasmic sperm injection (ICSI) without oocyte activation in two patients with different types of round-headed spermatozoa (globozoospermia) are reported. After controlled ovarian hyperstimulation and oocyte pick-up, retrieved oocytes were underwent ICSI without oocyte activation and a 33.33% (4/12) fertilisation rate was obtained in the first case, whereas an abnormal fertilisation was achieved in the second case. The transfer of two grade II embryos in the first couple resulted in clinical pregnancy with a healthy livebirth. It was concluded that the main problem of cases with globozoospermia was a low fertilisation rate or failure fertilisation, and even though ICSI and artificial oocyte activation have been employed to increase this rate, it is not necessarily needed to achieve a pregnancy.

The immune response of turbot, Scophthalmus maximus (L.), skin to high water temperature.

The immune responses of mucus from the skin of turbot, Scophthalmus maximus, were studied in relation to changing water temperature. Groups of fish were exposed to a programmed increase of 3 °C per 48 h, until the experimental water temperatures of 16, 20, 23, 25, 27 or 28 °C were reached. After 48 h at the relevant temperature, the expressions of immune-related factors were determined, including immunoglobulin M (IgM), IL-1ß, hepcidin, transferrin, lysozyme, acid/alkaline phosphatase and superoxide dismutase using RT-PCR and spectrophotometric methods. Significant changes in mucus immunity were observed, which paralleled with those previously reported for serum in other fish species. Hence, it is suggested that the serum and mucus immune system of turbot have a similar regulatory system. This information could be useful in better understanding the role of the mucus as a component of the innate immune system.

New modeling of reflection interference contrast microscopy including polarization and numerical aperture effects: application to nanometric distance measurements and object profile reconstruction.

We have developed a new and improved optical model of reflection interference contrast microscopy (RICM) to determine with a precision of a few nanometers the absolute thickness h of thin films on a flat surface in immersed conditions. The model takes into account multiple reflections between a planar surface and a multistratified object, finite aperture illumination (INA), and, for the first time, the polarization of light. RICM intensity I is typically oscillating with h. We introduce a new normalization procedure that uses the intensity extrema of the same oscillation order for both experimental and theoretical intensity values and permits us to avoid significant error in the absolute height determination, especially at high INA. We also show how the problem of solution degeneracy can be solved by taking pictures at two different INA values. The model is applied to filled polystyrene beads and giant unilamellar vesicles of radius 10-40 microm sitting on a glass substrate. The RICM profiles I(h) can be fitted for up to two to three oscillation orders, and extrema positions are correct for up to five to seven oscillation orders. The precision of the absolute distance and of the shape of objects near a substrate is about 5 nm in a range from 0 to 500 nm, even under large numerical aperture conditions. The method is especially valuable for dynamic RICM experiments and with living cells where large illumination apertures are required.

Molecular cloning and mRNA expression profiling of the first specific jasmonate biosynthetic pathway gene allene oxide synthase from Hyoscyamus niger.

In the endeavor to enhance the production of pharmaceutically valuable tropane alkaloids including hyoscyamine and scopolamine in Hyoscyamus niger, methyl jasmonate (MeJA) showed significant stimulation both in tropane biosynthetic pathway enzymes activities and tropane alkaloids yields. Therefore it was speculated that genetic engineering of jasmonate biosynthetic pathway might enhance the endogenous jasmonate concentration, followed by stimulating the production of tropane alkaloids. Herein a full-length cDNA encoding allene oxide synthase (AOS, EC 4.2.1.92), the first committed step enzyme in jasmonate biosynthetic pathway was reported (named HnAOS, GenBank accession: EF532599). HnAOS was a novel member of the cytochrome P450 (CYP74A) subfamily. Real-time quantitative PCR analysis showed that HnAOS mRNA accumulated mainly in stems, and responded significantly to wounding or methyl jasmonate.