Lipid peroxidation and biochemical parameters in maternal pre-delivery and post-delivery plasma

Lipid peroxidation (LPX) can play an important role in development of functional and pathological changes of maternal tissues in the course of pregnancy and delivery. LPX products were measured as thiobarbituric acid reacting substances (TBARS), using malondialdehyde as the standard solution. Actual TBARS determined in maternal post-delivery plasma (2.71 ± 0.602 nmol/mL) were not statistically different from those determined in pre-delivery plasma (3.45 ± 0.530 nmol/mL). TBARS production was measured in vitro in the both incubated plasma (30 min, 37°C) with and without the added LPX activator (125 μM L-ascorbate plus 5 μM FeSO₄). A difference in the TBARS formation was found only in the post-delivery plasma, as a result of approximately twice higher (marginally significant) TBARS formation in the incubated plasma without the added LPX activator comparing with the actual TBARS levels in this plasma. These results suggest that changes in maternal tissues in the process of labour could create suitable conditions for activation of LPX in maternal plasma. On the other hand, all other analysed biochemical parameters (iron, total iron-binding capacity, uric acid, proteins, magnesium, calcium, phosphate, glucose, potassium, sodium, chlorides, alanine aminotransferase, aspartate aminotransferase, γ-glutamyltransferase, creatine kinase, lactate dehydrogenase, α-hydroxybutyrate dehydrogenase, α-amylase, alkaline phosphatase, acid phosphatase in the post-delivery plasma were not different from those analysed in the pre-delivery plasma.     

Famotidine inhibits toll-like receptor 3-mediated inflammatory signaling in SARS-CoV-2 infection

Apart from prevention using vaccinations, the management options for COVID-19 remain limited. In retrospective cohort studies, use of famotidine, a specific oral H2 receptor antagonist (antihistamine), has been associated with reduced risk of intubation and death in patients hospitalized with COVID-19. In a case series, nonhospitalized patients with COVID-19 experienced rapid symptom resolution after taking famotidine, but the molecular basis of these observations remains elusive. Here we show using biochemical, cellular, and functional assays that famotidine has no effect on viral replication or viral protease activity. However, famotidine can affect histamine-induced signaling processes in infected Caco2 cells. Specifically, famotidine treatment inhibits histamine-induced expression of Toll-like receptor 3 (TLR3) in SARS-CoV-2 infected cells and can reduce TLR3-dependent signaling processes that culminate in activation of IRF3 and the NF-κB pathway, subsequently controlling antiviral and inflammatory responses. SARS-CoV-2-infected cells treated with famotidine demonstrate reduced expression levels of the inflammatory mediators CCL-2 and IL6, drivers of the cytokine release syndrome that precipitates poor outcome for patients with COVID-19. Given that pharmacokinetic studies indicate that famotidine can reach concentrations in blood that suffice to antagonize histamine H2 receptors expressed in mast cells, neutrophils, and eosinophils, these observations explain how famotidine may contribute to the reduced histamine-induced inflammation and cytokine release, thereby improving the outcome for patients with COVID-19.     

Transient receptor potential channel polymorphisms are associated with the somatosensory function in neuropathic pain patients

Transient receptor potential channels are important mediators of thermal and mechanical stimuli and play an important role in neuropathic pain. The contribution of hereditary variants in the genes of transient receptor potential channels to neuropathic pain is unknown. We investigated the frequency of transient receptor potential ankyrin 1, transient receptor potential melastin 8 and transient receptor potential vanilloid 1 single nucleotide polymorphisms and their impact on somatosensory abnormalities in neuropathic pain patients. Within the German Research Network on Neuropathic Pain (Deutscher Forscbungsverbund Neuropathischer Schmerz) 371 neuropathic pain patients were phenotypically characterized using standardized quantitative sensory testing. Pyrosequencing was employed to determine a total of eleven single nucleotide polymorphisms in transient receptor potential channel genes of the neuropathic pain patients and a cohort of 253 German healthy volunteers. Associations of quantitative sensory testing parameters and single nucleotide polymorphisms between and within groups and subgroups, based on sensory phenotypes, were analyzed. Single nucleotide polymorphisms frequencies did not differ between both the cohorts. However, in neuropathic pain patients transient receptor potential ankyrin 1 710G>A (rs920829, E179K) was associated with the presence of paradoxical heat sensation (p = 0.03), and transient receptor potential vanilloid 1 1911A>G (rs8065080, I585V) with cold hypoalgesia (p = 0.0035). Two main subgroups characterized by preserved (1) and impaired (2) sensory function were identified. In subgroup 1 transient receptor potential vanilloid 1 1911A>G led to significantly less heat hyperalgesia, pinprick hyperalgesia and mechanical hypaesthesia (p = 0.006, p = 0.005 and p<0.001) and transient receptor potential vanilloid 1 1103C>G (rs222747, M315I) to cold hypaesthesia (p = 0.002), but there was absence of associations in subgroup 2. In this study we found no evidence that genetic variants of transient receptor potential channels are involved in the expression of neuropathic pain, but transient receptor potential channel polymorphisms contributed significantly to the somatosensory abnormalities of neuropathic pain patients.     

Epileptiform activity in alcohol dependent patients and possibilities of its indirect measurement

Background

Alcohol dependence during withdrawal and also in abstinent period in many cases is related to reduced inhibitory functions and kindling that may appear in the form of psychosensory symptoms similar to temporal lobe epilepsy frequently in conditions of normal EEG and without seizures. Because temporal lobe epileptic activity tend to spread between hemispheres, it is possible to suppose that measures reflecting interhemispheric information transfer such as electrodermal activity (EDA) might be related to the psychosensory symptoms.

Methods and Findings

We have performed measurement of bilateral EDA, psychosensory symptoms (LSCL-33) and alcohol craving (ACQ) in 34 alcohol dependent patients and 32 healthy controls. The results in alcohol dependent patients show that during rest conditions the psychosensory symptoms (LSCL-33) are related to EDA transinformation (PTI) between left and right EDA records (Spearman r = 0.44, p<0.01).

Conclusions

The result may present potentially useful clinical finding suggesting a possibility to indirectly assess epileptiform changes in alcohol dependent patients.     

Assessing Risk Factors for Migraine: Differences in Gender Transmission

Aim

Our aim was to assess which specific factors are contributing to an increased risk of migraine in a group of 131 Portuguese families.

Methods

We studied 319 first-degree relatives, using a multilevel approach to account for the dependency among members from the same family. We included in the model relative’s gender, the proband’s gender and age-at-onset, to evaluate if any of these variables were associated with relative’s affection status. We also included in the model proband’s migraine subtype. We further assessed female and male transmissions within the proband nuclear family.

Results

Relatives’ gender was found to be a risk factor for migraine (Odds Ratio = 2.86; 95% CI = 1.75–4.67), with females at a higher risk. When splitting probands according to their migraine subtype, we found that none of the variables studied contributed to relatives of MA-probands affection-status. Our results also show a significant difference between proband’s transmission and the gender of the parents and offspring.

Conclusions

With this study, we showed that gender is truly a risk factor for migraine and that a gender-biased transmission is also observed. This reinforce the importance of identifying genes associated with migraine that are modulated by genes located in the sex chromosomes and the study of mitochondrial DNA or X-chromosome and hormonal-related effects associated with migraine susceptibility.     

The role of probiotics and natural bioactive compounds in modulation of the common molecular pathways in pathogenesis of atherosclerosis and cancer

Atherosclerosis and cancer are ranked among the most serious health problems in human medicine. Various predictive and etiological factors, biomarkers and molecular pathways of disease development and progression common to atherosclerosis and cancer suggest that the two most common diseases in worldwide dimension are far more closely aligned than previously believed. It is hypothesized that atherosclerosis and cancer are variants of a similar disease process. Shared disease progression in atherosclerosis and cancer is the emergence of similar novel approaches to therapy. On previous knowledge, it may be hypothesized that not only common approaches to therapy but also preventive strategies could be efficacious in both diseases. The results of in vitro and in vivo animal experiments, clinical and epidemiological studies and also the results of our experiments using animal experimental models of atherosclerosis and carcinogenesis indicate that probiotics, prebiotics, plants and their extracts and poly-unsaturated fatty acids could be effectively used in prevention of both atherosclerosis and colorectal cancer and decrease the disease risk. Future research should answer the question whether probiotic microorganisms and natural bioactive substances could effectively influence the molecular mechanisms in pathogenesis of atherosclerosis and cancer.     

Co-stimulatory and adhesion molecules of dendritic cells in rheumatoid arthritis

Dendritic cells (DCs) in the rheumatoid arthritis (RA) joint mediate the immunopathological process and act as a potent antigen presenting cell. We compared the expression of co-stimulatory and adhesion molecules on DCs in RA patients versus controls with traumatic joint lesions and evalulated the correlation between the immunophenotypical presentation of DCs and the clinical status of the disease. Samples of peripheral venous blood, synovial fluid (SF) and synovial tissue (ST) were obtained from 10 patients with RA at the time of hip or knee replacement and from 9 control patients with knee arthroscopy for traumatic lesions. Clinical status was appreciated using the DAS28 score. Blood, SF and dissociated ST cell populations were separated by centrifugation and analyzed by flow cytometry. Cells phenotypes were identified using three-color flow cytometry analysis for the following receptors HLA-DR, CD80, CD83, CD86, CD11c, CD18, CD54, CD58, CD3, CD4, CD8, CD19, CD20, CD14, CD16, CD56. HLA-DR molecules, co-stimulatory receptors CD80, CD86, CD83 and adhesion molecules CD18, CD11c, CD54, CD58, were analyzed by two-color immunofluorescence microscopy on ST serial sections. In patients with active RA (DAS28>5.1) we found a highly differentiated subpopulation of DCs in the ST and SF that expressed an activated phenotype (HLA-DR, CD86+, CD80+, CD83+, CD11c+, CD54+, CD58+). No differences were found between circulating DCs from RA patients and control patients. Our data suggest an interrelationship between clinical outcome and the immunophenotypical presentation of DCs. Clinical active RA (DAS28>5.1) is associated with high incidence of activated DCs population in the ST and SF as demonstrated by expression of adhesion and co-stimulatory molecules.     

A new role in hemostasis for the adhesion receptor P-selectin

The adhesion receptor P-selectin has long been known to support leukocyte rolling and emigration at sites of inflammation. Recently, P-selectin was also revealed to be a key molecule in hemostasis and thrombosis, mediating platelet rolling, generating procoagulant microparticles containing active tissue factor and enhancing fibrin deposition. Elevated levels of plasma P-selectin are indicative of thrombotic disorders and predictive of future cardiovascular events. Because the interaction between P-selectin and its receptor P-selectin glycoprotein ligand-1 (PSGL-1) represents an important mechanism by which P-selectin induces the formation of procoagulant microparticles and recruits the microparticles to thrombi, anti-thrombotic strategies are currently aimed at inhibiting this interaction. Recent developments also suggest that the procoagulant potential of P-selectin could be used to treat coagulation disorders such as hemophilia A.     

Fibronectin-coated beads are endocytosed by cells and align with microfilament bundles

Carboxylated latex beads coated with fibronectin bind to the surfaces of NIL8 hamster cells. Similar beads coated with bovine serum albumin, gelatin or normal rabbit immunoglobulin do not bind to the cells, whereas beads coated with polylysine or rabbit anti-hamster immunoglobulin bind, as do fibronectin-coated beads. Surface-bound beads are endocytosed by the cells. This endocytosis is inhibited by N-ethyl maleimide or low temperatures. The endocytosed beads form arrays aligned with the microfilament bundles inside the cells and after extended incubations (18–24 h) become concentrated in the perinuclear region. These results suggest that, after phagocytosis of large particles, the endocytic vesicles align with microfilament bundles, as previously reported for coated pits involved in receptor-mediated endocytosis. The system described here offers possibilities for the analysis of membrane and transmembrane interactions involved in cell-substratum binding and phagocytosis.