The Toxicity of Wiped Dust and Airborne Microbes in Individual Classrooms Increase the Risk of Teachers’ Work-Related Symptoms: A Cross-Sectional Study

Background: The causes and pathophysiological mechanisms of building-related symptoms (BRS) remain open. Objective: We aimed to investigate the association between teachers’ individual work-related symptoms and intrinsic in vitro toxicity in classrooms. This is a further analysis of a previously published dataset. Methods: Teachers from 15 Finnish schools in Helsinki responded to the symptom survey. The boar sperm motility inhibition assay, a sensitive indicator of mitochondrial dysfunction, was used to measure the toxicity of wiped dust and cultured microbial fallout samples collected from the teachers’ classrooms. Results: 231 teachers whose classroom toxicity data had been collected responded to the questionnaire. Logistic regression analysis adjusted for age, gender, smoking, and atopy showed that classroom dust intrinsic toxicity was statistically significantly associated with the following 12 symptoms reported by teachers (adjusted ORs in parentheses): nose stuffiness (4.1), runny nose (6.9), hoarseness (6.4), globus sensation (9.0), throat mucus (7.6), throat itching (4.4), shortness of breath (12.2), dry cough (4.7), wet eyes (12.7), hypersensitivity to sound (7.9), difficulty falling asleep (7.6), and increased need for sleep (7.7). Toxicity of cultured microbes was found to be associated with nine symptoms (adjusted ORs in parentheses): headache (2.3), nose stuffiness (2.2), nose dryness (2.2), mouth dryness (2.8), hoarseness (2.2), sore throat (2.8), throat mucus (2.3), eye discharge (10.2), and increased need for sleep (3.5). Conclusions: The toxicity of classroom dust and airborne microbes in boar sperm motility inhibition assay significantly increased teachers’ risk of work-related respiratory and ocular symptoms. Potential pathophysiological mechanisms of BRS are discussed.

Impact of Immunoglobulin Isotype and Epitope on the Functional Properties of Vibrio cholerae O-Specific Polysaccharide-Specific Monoclonal Antibodies

ABSTRACT Vibrio cholerae causes the severe diarrheal disease cholera. Clinical disease and current oral cholera vaccines generate antibody responses associated with protection. Immunity is thought to be largely mediated by lipopolysaccharide (LPS)-specific antibodies, primarily targeting the O-antigen. However, the properties and protective mechanism of functionally relevant antibodies have not been well defined. We previously reported on the early B cell response to cholera in a cohort of Bangladeshi patients, from which we characterized a panel of human monoclonal antibodies (MAbs) isolated from acutely induced plasmablasts. All antibodies in that previous study were expressed in an IgG1 backbone irrespective of their original isotype. To clearly determine the impact of affinity, immunoglobulin isotype and subclass on the functional properties of these MAbs, we re-engineered a subset of low- and high-affinity antibodies in different isotype and subclass immunoglobulin backbones and characterized the impact of these changes on binding, vibriocidal, agglutination, and motility inhibition activity. While the high-affinity antibodies bound similarly to O-antigen, irrespective of isotype, the low-affinity antibodies displayed significant avidity differences. Interestingly, despite exhibiting lower binding properties, variants derived from the low-affinity MAbs had comparable agglutination and motility inhibition properties to the potently binding antibodies, suggesting that how the MAb binds to the O-antigen may be critical to function. In addition, not only pentameric IgM and dimeric IgA, but also monomeric IgA, was remarkably more potent than their IgG counterparts at inhibiting motility. Finally, analyzing highly purified F(ab) versions of these antibodies, we show that LPS cross-linking is essential for motility inhibition. IMPORTANCE Immunity to the severe diarrheal disease cholera is largely mediated by lipopolysaccharide (LPS)-specific antibodies. However, the properties and protective mechanisms of functionally relevant antibodies have not been well defined. Here, we have engineered low and high-affinity LPS-specific antibodies in different immunoglobulin backbones in order to assess the impact of affinity, immunoglobulin isotype, and subclass on binding, vibriocidal, agglutination, and motility inhibition functional properties. Importantly, we found that affinity did not directly dictate functional potency since variants derived from the low-affinity MAbs had comparable agglutination and motility inhibition properties to the potently binding antibodies. This suggests that how the antibody binds sterically may be critical to function. In addition, not only pentameric IgM and dimeric IgA, but also monomeric IgA, was remarkably more potent than their IgG counterparts at inhibiting motility. Finally, analyzing highly purified F(ab) versions of these antibodies, we show that LPS cross-linking is essential for motility inhibition.

Dysfunction of Shh signaling activates autophagy to inhibit trophoblast motility in recurrent miscarriage

In early pregnancy, the placenta anchors the conceptus and supports embryonic development and survival. This study aimed to investigate the underlying functions of Shh signaling in recurrent miscarriage (RM), a serious disorder of pregnancy. In the present study, Shh and Gli2 were mainly observed in cytotrophoblasts (CTBs), Ptch was mainly observed in syncytiotrophoblasts (STBs), and Smo and Gli3 were expressed in both CTBs and STBs. Shh signaling was significantly impaired in human placenta tissue from recurrent miscarriage patients compared to that of gestational age-matched normal controls. VEGF-A and CD31 protein levels were also significantly decreased in recurrent miscarriage patients. Furthermore, inhibition of Shh signaling impaired the motility of JAR cells by regulating the expression of Gli2 and Gli3. Intriguingly, inhibition of Shh signaling also triggered autophagy and autolysosome accumulation. Additionally, knockdown of BECN1 reversed Gant61-induced motility inhibition. In conclusion, our results showed that dysfunction of Shh signaling activated autophagy to inhibit trophoblast motility, which suggests the Shh pathway and autophagy as potential targets for RM therapy.

Searching for New Tools to Counteract the Helicobacter pylori Resistance: The Positive Action of Resveratrol Derivatives

The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to improve the eradication rate including alternative treatments combining antibiotic and non-antibiotic compounds with synergistic action. In this study, the antibacterial (MIC/MBC) and anti-virulence effects (biofilm reduction and swarming motility inhibition) of resveratrol-RSV and new synthetized RSV-phenol derivatives, with a higher bioavailability, alone and combined with levofloxacin-LVX were evaluated against resistant H. pylori clinical strains. The experiments were confirmed in vivo using the Galleria mellonella model. Among the studied RSV derivatives, RSV-3 and RSV-4 possessed higher antibacterial activity with respect to RSV (MICs from 6.25 to 200 µg/mL and from 3.12 to 200 µg/mL, respectively). RSV, RSV-3, and RSV-4 were able to synergize with LVX restoring its effect in two out of seven clinical resistant strains tested for the study. RSV, RSV-3, and RSV-4, alone and with LVX at sub-MIC and sub-synergistic concentrations, significantly reduced the biofilm formation. Moreover, RSV-3 and RSV-4 reduced the H. pylori swarming motility on soft agar. RSV, RSV-3, and RSV-4 were non-toxic for G. mellonella larvae and displayed a protective effect against H. pylori infection. Overall, RSV–phenol derivatives should be considered interesting candidates for innovative therapeutic schemes to tackle the H. pylori antibiotic resistance.

Dysfunction of Shh signaling impaired trophoblast motility and autophagy is involved in poor placentation of recurrent miscarriage

Background: In early pregnancy, the placenta anchors the conceptus and supports embryonic development and survival. This study aimed to investigate the underlying functions of Shh signaling on recurrent miscarriage, an serious disorder of pregnancy. Methods: Immunofluorescence and immunohistochemistry were used to detect protein expression and its location in placental tissues. Quantitative real-time RT-PCR and Western blot analysis were performed to examine mRNA and protein levels, respectively. Lentiviruses expressing short hairpin RNA were used to knock down the target genes. Cell invasion and migration were performed by with or without Matrigel-coated transwell, respectively. Primary trophoblast migration was performed by villous explant assay. RNA-sequence was used to investigate the genes transcription profile. CCK-8 assay was used to evaluate cell viability. Flow cytometry was used to evaluate cell apoptosis. Results: Our results showed that Shh and Gli2 were mainly located in cytotrophoblasts (CTBs), Ptch was mainly located in syncytiotrophoblasts (STBs), while Smo and Gli3 were expressed in both CTBs and STBs. Compared to the gestational age-matched normal human placenta, the expression of Shh was significantly decreased in recurrent miscarriage. Furthermore, inhibition of Shh signaling impaired motility of JAR cells via regulating the expression of Gli2 and Gli3 to decrease AKT Ser473 phosphorylation, elevate E-cadherin and VEGFA. Intriguingly, inhibition of Shh signaling also enhanced autophagy and autolysosome. Additionally, knockdown BECN1 reversed the effect of Gant61 on motility inhibition. Conclusion: Our results indicated that dysfunction of Shh signaling impaired trophoblast motility, angiogenesis and activated autophagy in villous trophoblast, which would contribute to the pathophysiology of recurrent miscarriage.

Composition Analysis and Anti-Cancer Properties of Two Endemic Phlomis Species (Phlomis cypria Post and Phlomis brevibracteata Turrill) from Cyprus

Recently, different herbal compounds have been studied for their curative properties against Hepatocellular Carcinoma (HCC). There are over 100 Phlomis species native to the Mediterranean region, which have pharmacological activities against gastrointestinal system and liver diseases. Phlomis brevibracteata Turrill (PBT) and Phlomis cypria Post (PCP) are endemic plants of North-Cyprus belonging to the Lamiaceae family. In this study, chemical composition of the 70% methanol extracts of Phlomis species were analyzed by LC/MS/MS and antiradical activities were evaluated by DPPH● and ABTS+● radical scavenging activity tests. Anti-carcinogenic activities on HCC cell lines were investigated using MTT and wound healing assays, intercellular ROS scavenging activities were tested by DCFH-DA assay. Forsytoside B and caffeoylquinic acids were found as the main phenolic compounds which are possibly responsible for antiradical activities of the plant extracts. Significant decrease in cell viability was noticed for both extracts at concentrations over 1000 µg/mL. PBT treatment also resulted in cell motility inhibition. Additionally, both basal and induced oxidative states of all cell lines significantly were decreased by PBT. Since extracts showed cytotoxic, antioxidant and motility inhibitory activities, Phlomis species can be a good candidate for further studies with the goal of new anticancer chemotherapeutic discoveries.