Gasoline fume inhalation induces apoptosis, inflammation, and favors Th2 polarization in C57BL/6 mice.

Gasoline exposure has been widely reported in the literature as being toxic to human health. However, the exact underlying molecular mechanisms triggered by its inhalation have not been thoroughly investigated. We herein present a model of sub-chronic, static gasoline vapor inhalation in adult female C57BL/6 mice. Animals were exposed daily to either gasoline vapors (0.86 g/animal/90 min) or ambient air for 5 days/week over 7 consecutive weeks. At the end of the study period, toxic and molecular mechanisms underlying the inflammatory, oxidative, and apoptotic effects triggered by gasoline vapors, were examined in the lungs and liver of gasoline-exposed (GE) mice. Static gasoline exposure induced a significant increase (+21%) in lungs/body weight (BW) ratio in GE versus control (CON) mice along with a pulmonary inflammation attested by histological staining. The latter was consistent with increases in the transcript levels of proinflammatory cytokines [Interleukins (ILs) 4 and 6], respectively by ~ 6- and 4-fold in the lungs of GE mice compared to CON. Interestingly, IL-10 expression was also increased by ~ 10-fold in the lungs of GE mice suggesting an attempt to counterbalance the established inflammation. Moreover, the pulmonary expression of IL-12 and TNF-α was downregulated by 2- and 4-fold, respectively, suggesting the skewing toward Th2 phenotype. Additionally, GE mice showed a significant upregulation in Bax/Bcl-2 ratio, caspases 3, 8, and 9 with no change in JNK expression in the lungs, suggesting the activation of both intrinsic and extrinsic apoptotic pathways. Static gasoline exposure over seven consecutive weeks had a minor hepatic portal inflammation attested by H&E staining along with an increase in the hepatic expression of the mitochondrial complexes in GE mice. Therefore, tissue damage biomarkers highlight the health risks associated with vapor exposure and may present potential therapeutic targets for recovery from gasoline intoxication.

Evaluation of waterpipe smoke toxicity in C57BL/6 mice model.

Waterpipe smoking is a popular pastime worldwide with statistics pointing to an alarming increase in consumption. In the current paper, the evaluation of sub-chronic waterpipe smoke exposure was undertaken using C57BL/6 female mice using a dynamic exposure setting to emulate smoke exposure. Mice were daily subjected to either one (single exposure, SE) or two sessions (double exposure, DE) of waterpipe-generated smoke (two-apple flavor) for a period of two months. Although lungs histopathological examination pointed to a minor inflammation in smoke-exposed mice compared to control air-exposed (CON) group, the lung weights of the waterpipe-exposed mice were significantly higher (+72% in SE and +39% in DE) (p < 0.01) when compared to CON group. Moreover, changes in the protein expression of several proteins such as iNOS and JNK were noted in the lungs of smoke-exposed mice. However, no changes in p38 and EGFR protein levels were noted between the three groups of mice. Our results mainly showed a significant increase in urea serum levels (+28%) in SE mice along with renal pathological damage in both SE and DE mice compared to CON. Additionally, severe significant DNA damages (p < 0.05) were reported in the lungs, kidneys, bone marrow and liver of waterpipe-exposed animals, using MTS and COMET assays. These findings highlighted the significant risks posed by sub-chronic waterpipe smoke exposure in the selected animal model and the pressing need for future better management of waterpipe indoor consumption.

Curcumin encapsulated colloidal amphiphilic block co-polymeric nanocapsules: colloidal nanocapsules enhance photodynamic and anticancer activities of curcumin.

Curcumin-based novel colloidal nanocapsules were prepared from amphiphilic poly(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethylene oxide) (F108). These colloidal nanocapsules appeared as spherical particles with size ranging between 270 and 310 nm. Curcumin fluorescence spectra exhibited an aggregation-induced 23 nm red-shift of the emission maximum in addition to the enhancement of the fluorescence quantum yield in these nanocapsules. The cytotoxicity of curcumin and colloidal nanocapsules was assessed using human derived immortalized cell lines (A549 and A375 cells) in the presence and absence of light irradiation. The nanocapsules exhibited a >30-fold decrease in IC50, suggesting enhanced anticancer activity associated with curcumin encapsulation. Higher toxicity was also reported in the presence of light irradiation (as shown by the IC50 data), indicating their potential for future application in photodynamic therapy. Finally, A375 cells treated with curcumin and the nanocapsules showed a significant increase in single- and/or double-strand DNA breaks upon exposure to light, indicating promising biological effects.

Characterization and cytotoxicity assessment of nargile smoke using dynamic exposure.

Objectives: Nargile (waterpipe) smoking has gained popularity in the Middle East and throughout the world. In this research, a new dynamic methodology was conceived. This methodology was deployed for direct in vitro assessment of cytotoxicity, genotoxicity, and apoptotic potential of smoke generated from a single nargile session. Materials and methods: A549 cells were deployed in a designed system to assess the cytotoxicity of generated smoke. The smoke was characterized using Gas chromatography-mass spectrometry (GC-MS) profiling for major organic compounds, whereas the remaining chemical and physical parameters were tabulated from published data. The cytoxicity of smoke generated from five commercial flavored tobacco products was assessed using the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2-H-tetrazolium) (MTS) assay. The genotoxicity was also measured using the comet assay, while apopoptosis was evaluated using Annexin V/propidium iodide staining.Results: The data indicated acute cytotoxicity emanating from smoke products in all tested tobacco flavors. Significant loss of viability and mitochondrial activity was observed 40 min post smoke exposure (Double-Apple flavored), while DNA damage onset was reported as early as 20 min of exposure. Microscopical analysis showed a systematic increase in cell rounding post exposure indicating cellular loss of adhesion and potential membrane damages. Finally, the Annexin V/propidium iodide cellular staining showed signs of late apoptosis or necrosis in exposed cells. Conclusions: The presented data clearly indicated significant in vitro cytotoxicity, genotoxicity and apoptosis/necrosis associated with a 60-min single session of nargile smoking.

[Urinary incontinence after radical prostatectomy for prostate cancer-data from 17,149 patients from 125 certified centers]. / Harninkontinenz nach radikaler Prostatektomie beim Prostatakarzinom ­ aktuelle Daten von 17.149 Patienten aus 125 zertifizierten Zentren.

BACKGROUND: In addition to erectile dysfunction, urinary incontinence is the most common functional limitation after radical prostatectomy (RPE) for prostate cancer (PCa). The German S3 guideline recommends informing patients about possible effects of the therapy options, including incontinence. However, only little data on continence from routine care in German-speaking countries after RPE are currently available, which makes it difficult to inform patients. OBJECTIVE: The aim of this work is to present data on the frequency and severity of urinary incontinence after RPE from routine care. MATERIALS AND METHODS: Information from the PCO (Prostate Cancer Outcomes) study is used, which was collected between 2016 and 2022 in 125 German Cancer Society (DKG)-certified prostate cancer centers in 17,149 patients using the Expanded Prostate Cancer Index Composite Short Form (EPIC-26). Changes in the "incontinence" score before (T0) and 12 months after RPE (T1) and the proportion of patients who used pads, stratified by age and risk group, are reported. RESULTS: The average score for urinary incontinence (value range: 0-worst possible to 100-best possible) was 93 points at T0 and 73 points 12 months later. At T0, 97% of the patients did not use a pad, compared to 56% at T1. 43% of the patients who did not use a pad before surgery used at least one pad a day 12 months later, while 13% use two or more. The proportion of patients using pads differs by age and risk classification. CONCLUSION: The results provide a comprehensive insight into functional outcome 12 months after RPE and can be taken into account when informing patients.

Characterization of cytotoxicity of airborne particulates from urban areas of Lahore.

A number of species (organic and inorganic) in airborne particulates cause the toxicity to living being. The potential of in vitro test methods were explored for toxicity assessment of trace toxic elements (inorganic species) present in ambient air on human being (lungs). A year long sampling of airborne particles (PM2.5) was carried (April 2008 to March 2009) in Lahore, Pakistan. A total of thirty nine samples were collected on 47 mm Zefluor Teflon filter membranes and each was analysed to characterize for the elements: Sb, As, Be, Cd, Cr, Co, Pb, Mn, Hg using ICP-MS in water extract and total acid digestate. The samples cytotoxicity was also established using lung derived cells and MTS colorimetric assays. This generated dose response curves and IC50 values for the elemental mixtures identified on the Teflon filter membrane. The results indicated that even at low concentrations airborne elemental mixtures displayed an additive toxic effect.

Acute cytotoxicity, genotoxicity, and apoptosis induced by petroleum VOC emissions in A549 cell line.

Gasoline is an essential petroleum-derived product powering the automotive economy worldwide. This research focused on the Volatile Organic Component (VOC) cocktail resulting from gasoline evaporation. Petroleum fugitive VOC inhalation by petrol station attendants have been widely associated with toxicological and health risks concerns. Another unusual practice in poor nations is gasoline sniffing to get high which can lead to intoxication and organ damages. In this study, a static air/liquid interface methodology was designed to emulate acute human lung-derived cell exposure to all the gasoline-derived generated VOCs. The research investigated the cytotoxic and genotoxic end points resulting from whole gasoline fumes in vitro exposure using A549 cells. Petroleum-derived VOCs were identified and characterized by GC-MS. VOCs exposure was emulated in a controlled environment by evaporating spiked crude gasoline (1 to 100 µl) in a closed exposure chamber. In the chamber, A549 cultured cells on snapwell inserts were exposed on their apical side to various concentrations of generated vapors for one hour at 37 °C to mimic lung exposure. The results indicated that acute gasoline whole VOCs exposure reduced cell viability (IC50 = 485 ppm immediately and IC50 = 516 ppm 24 h post-exposure), disrupted cell membrane integrity though LDH leakage and induced DNA damages. Furthermore, VOC exposure triggered caspase-independent apoptosis in exposed cells through upregulation of apoptotic pathways. Overall, the presented findings generated by the static exposure technique showed a practical and reproducible model that can be used to assess acute crude VOCs mixture toxicity endpoints and cell death pathways.

Novel Platinum(II) and Platinum(IV) Antitumor Agents that Exhibit Potent Cytotoxicity and Selectivity.

A novel platinum(II) complex 47OMESS(II) and its platinum(IV) derivative 47OMESS(IV) were synthesized and characterized. Cytotoxicity studies against mesenchymal cells (MCs) and lung (A549), breast (MDA-MB-231), and melanoma (A375) cancer cells demonstrated 7-20-fold superior activity for both complexes relative to cisplatin. Remarkably, 47OMESS(IV) demonstrated 17-22-fold greater selectivity toward the cancerous cells compared to the non-cancerous MCs. Western blot analysis on A549 cells showed the involvement of the intrinsic apoptotic pathway. Cellular fractionation and uptake experiments in A549 cells using ICP-mass spectrometry (MS) indicated that 47OMESS(II) and 47OMESS(IV) cross the cellular membrane predominantly via active transport mechanisms. The significant improvement in selectivity that is exhibited by 47OMESS(IV) is reported for the first time for this class of complexes.

E-cigarette aerosol induced cytotoxicity, DNA damages and late apoptosis in dynamically exposed A549 cells.

In this study, the acute toxicological impacts associated with electronic cigarettes consumption were determined using a novel dynamic exposure methodology. The methodology was deployed to test various e-cigarette generated aerosols in A549 cell cultures. The e-liquid chemical profiling was achieved using GC-MS analysis while toxicity of diluted e-liquids aerosols was reported using numerous cytotoxicity assays. The presented findings pointed to acute aerosol exposure (thirty puffs at 40 W of power and higher) inducing significant cytotoxic, genotoxic, and apoptotic induction in exposed cells. These findings highlighted the significant risks posed by e-cigarette usage. The proposed methodology proved to be a useful tool for future screening of e-liquids generated aerosols toxicity. Future research is needed to establish the chronic toxicity resulting from long-term e-cigarette consumption.

Purified c-phycoerythrin: safety studies in rats and protective role against permanganate-mediated fibroblast-DNA damage.

We have evaluated in vitro cytotoxicity of cyanobacterial phycoerythrin (C-PE) on three human cell lines by cell proliferation and neutral red uptake assays. No toxic effects of C-PE were observed to any of the cell lines tested. The protective role of purified C-PE to potassium permanganate-mediated human fibroblast-DNA damage was assessed by comet assay at 0 (control), 10 and 20 microg C-PE ml(-1) doses in pre-, simultaneous and post-mutagen exposure conditions. Significant DNA damage was detected only in post-mutagen exposure conditions. Our findings confirmed that the C-PE is non-toxic and provides protection against permanganate-mediated DNA damage. The preliminary acute (2000 mg C-PE kg(-1) body weight, b.w.) and 90 day sub-chronic (0, 5, 15 and 25 mg C-PE kg(-1) b.w./day) oral toxicity studies of purified C-PE in male albino rats showed no mortality or treatment-related major clinical signs, and all the doses of C-PE were well tolerated. The no observed adverse effect level and no observed effect level were found to be 15 and 5 mg C-PE kg(-1) b.w./day respectively.

Nicotine induces resilience to chronic social defeat stress in a mouse model of water pipe tobacco exposure by activating BDNF signaling.

The purpose of this study was to investigate how nicotine in the context of water pipe tobacco smoking (WTS) affects depression and anxiety-like behaviors associated with chronic social defeat stress (CSDS). Male C57BL/6 J mice were exposed to WTS or received intraperitoneal injections of nicotine for thirty days then subjected to CSDS for ten days. During CSDS, mice were exposed to WTS or received nicotine injections. The social interaction and open-field tests were used to classify animals as resilient or susceptible to stress and to evaluate their anxiety-like behavior. After behavioral testing, mice continued to be exposed to WTS/nicotine for ten days and their behavior was reexamined. The involvement of brain derived neurotrophic factor signaling in the nicotine-mediated effects was assessed with the tropomyosin receptor kinase B (TRKB) inhibitor, ANA-12. We found that WTS promotes resilience to stress and rescues social avoidance. Even though WTS initially decreased anxiety-like behaviors, prolonged exposure after the completion of CSDS significantly induced anxiety-like behaviors. Finally, we showed that nicotine mediates the effects of WTS only on resilience to stress by increasing BDNF and TRKB levels and signaling. Our results suggest that the pathways mediating resilience to stress and anxiety are distinct and that nicotine mediates the effects of WTS on social behavior, but not anxiety, by activating BDNF signaling. Significance statement: This study reports the positive effect of WTS and nicotine on social behavior. Furthermore, it shows the negative effects of prolonged WTS on anxiety-like behaviors and suggests that these effects are not necessarily mediated by nicotine. Finally, it identifies BDNF/TRKB signaling pathway as a major mediator of the positive effects of nicotine on social interaction. As a result, this work emphasizes the importance of considering the activation status of this signaling pathway when developing smoking cessation strategies.

Human skin explants an in vitro approach for assessing UVB induced damage.

Lifestyle changes involving frequent outdoor activities are contributing to higher exposure to harmful ultraviolet light (UVB). The acute effects of UVB irradiation on human skin was evaluated in this study using freshly excised human skin from elective surgery subjected to UVB doses (0-3.76 J/cm2). The assessment of UVB induced cellular and skin damages was undertaken at two time points immediately and 24 h post exposure using in vitro, and immunohistochemical staining techniques. The results indicated no significant loss of skin integrity or significant acute mitochondrial cellular damages in UVB exposed skin sections as measured by the MTS cytotoxicity assay. The other key markers of damage showed significant extracellular LDH membrane leakages and upregulation of inflammatory cytokines such as IL-1ß. Skin integrity analysis was also undertaken using H&E, HLADR, and anti-cytokeratin antibodies. The results showed significant epidermal changes, basal cell activation and Langerhans cells depletion. The research proved the usefulness of freshly excised human skin explant model in measuring UVB damage. Furthermore, freshly excised human skin maintains the natural layering and therefore does not pose the same challenges faced by commercially available reconstructed skin in terms of higher costs and accurate mimicking of all the complex interactions observed in human skin.

Municipal leachates health risks: Chemical and cytotoxicity assessment from regulated and unregulated municipal dumpsites in Lebanon.

The proper management of municipal waste is critical for resource recovery, sustainability and health. Lebanon main approach for managing its municipal waste consisted of landfill disposal with minimal recycling capacity. This approach contributed to exceeding the holding capacity of existing landfills leading eventually to their closures. The closure of a major landfill (Naameh landfill) servicing Beirut and Mount Lebanon areas led to municipal wastes piling in the streets and forests for more than a year in 2016. The main problem identified in the municipal wastes consisted of untreated leachates (from regulated and unregulated dumpsites) going straight into the Mediterranean Sea. Therefore leachate samples were collected and subjected to chemical characterization followed by biological assessment. The chemical characterization and profiling of the Lebanese leachates were compared to results reported in Lebanon, Europe and United States as well as to the toxicity reference values (TRV). The biological assessment was conducted in vitro using human derived immortalized cell cultures. This strategy revealed significant alarming cellular organelles and DNA damages using in vitro cytotoxicity assays (MTS and comet assay). The significant damages observed at the cellular level prompted further animal model investigations using BALB/c mice. The animal data pointed to significant upregulation of liver activity enzymes coupled with significant damage expression in liver spleen and bone marrow DNA. The presented research clearly indicated that there is an urgent need for development of national waste strategies for proper treatment and disposal of municipal waste leachates in Lebanon.

A long-lived cuprous bis-phenanthroline complex for the photodynamic therapy of cancer.

Copper is an earth-abundant and a biologically essential metal that offers a promising alternative to noble metals in photochemistry and photobiology. In this work, a series of sterically encumbered Cu(i) bis-phenanthroline complexes were investigated for their use in photochemotherapy (PCT). It was found that Cu(dsbtmp)2+ [dsbtmp = 2,9-disec-butyl-3,4,7,8-tetramethyl-1,10-phenanthroline] (compound 3), which possessed the longest excited state lifetime, exhibited significant in vitro photocytotoxicity on A375 (human malignant melanoma) and A549 (human lung carcinoma) cell lines. Fluorescence imaging demonstrated the significant uptake and localization of compound 3 in a perinuclear fashion. A comet assay indicated the induction of DNA damage in the dark. The DNA breaks were significantly amplified upon photoactivation. The light-induced enhancement of cytotoxicity was associated with the formation of reactive oxygen species (ROS), a known intermediate in photodynamic therapy (PDT). This successful demonstration of photocytotoxicity using long-lived cuprous phenanthroline paves the way to exploit this class of photosensitizers for PDT applications.

UVB damage onset and progression 24 h post exposure in human-derived skin cells.

The focus of this research was on UVB radiation (280-320 nm) responsible for cellular changes in skin of acute and chronically exposed individuals. This study investigated the acute cellular damages triggered by UVB exposure of cultured human fibroblasts and keratinocyte cells immediately and 24 h post exposure in order to understand damage onset and progression. The study evaluated a number of cellular parameters including mitochondria, lysosomes, cell membrane, DNA damages as well as pro and anti-apoptotic protein expression levels. Cellular organelle damages were assessed by a battery of in vitro toxicological assays using MTS and Neutral red cytotoxicity assays. Cell membrane damages were also assessed by measuring lactate dehydrogenase (LDH) enzyme leakage from UVB exposed cells. Lastly DNA damages was assessed using the comet assay while protein expression was evaluated using Western Blot. In this study we reported in all our assay systems (MTS, NR and LDH) that cellular damages were UVB dose dependent with damages amplified 24 h post exposure. Our results also indicated that incubation of exposed cells for a period of 24 h increased the sensitivity of the assay systems used. The increased sensitivity in detecting early cytotoxic damages was manifested though organelle damage measurement at very low doses which were not manifested immediately post exposure. The data also indicated that HaCaT cells were most sensitive in detecting UVB triggered damages immediately and 24 h post exposure using the MTS assay. We also established upregulation and downregulation of various apoptotic proteins at various time points post exposure. The presented data clearly indicated the need for a comprehensive assessment of UVB damages 4 and 24 h post exposure due to the different assay sensitivities in addition to various signaling mechanisms activated at different time points post exposure.

Is subjective hyperhidrosis assessment sufficient enough? prevalence of hyperhidrosis among young Polish adults.

The aim of this work was to assess the prevalence of palmar and axillary hyperhidrosis among young Polish adults. Additionally, this work aimed at comparing the subjective and objective (gravimetric) method of hyperhidrosis assessment. Healthy medical students, volunteering to take part in this study, were included. The participants filled out a questionnaire assessing the occurrence and subjective intensity of hyperhidrosis in different areas of the body. Additionally, the students were subjected to gravimetric assessment in four localizations: the face, palms, axillae and abdomino-lumbar area. Two hundred and fifty-three students (102 males and 151 females, mean age 24.3 ± 3.21 years) were included in the study. Forty-two (16.7%) participants declared that they suffer from hyperhidrosis. Out of the 42 students declaring any type of hyperhidrosis, only 20 (47.6%) exceeded the gravimetric reference values. From among the students that exceeded the normative values for palmar hyperhidrosis, only 10 (55.6%) were aware of their hyperhidrosis. In the group of students that exceeded the normative values for axillary hyperhidrosis, 16 (39%) were aware of their hyperhidrosis. Subjectively declared hyperhidrosis incidence may significantly exceed the real-life occurrence of this disease. Basing studies solely on data gathered from questionnaires, may lead to false results. It is imperative, when assessing patients suffering from hyperhidrosis, to use both objective and subjective methods of evaluation.