Comparative analysis of lipid-peptide nanoparticles prepared via microfluidics, reverse phase evaporation, and ouzo techniques for efficient plasmid DNA delivery.

In the current "era of lipid carriers," numerous strategies have been developed to manufacture lipid nanoparticles (LNPs). Nevertheless, the potential impact of various preparation methods on the characteristics, use, and/or stability of these LNPs remains unclear. In this work, we attempted to compare the effects of three different preparation methods: microfluidics (MF), reverse phase evaporation (RV), and ouzo (OZ) on lipid-peptide NPs (LPNPs) as plasmid DNA delivery carriers. These LPNPs had the same components, namely DOTMA cationic lipid, DSPC, cholesterol, and protamine. Subsequently, we compared the LPNPs in terms of their physicochemical features, functionality as gene delivery vehicles in two distinct cell lines (NT2 and D1-MSCs), and finally, their storage stability over a six-month period. It was clear that all three LPNP formulations worked to deliver EGFP-pDNA while keeping cells alive, and their physicochemical stability was high for 6 months. However, the preparation technique had a significant impact on their physicochemical characteristics. The MF produced LPNPs with a lesser size, polydispersity index, and zeta potential than the other synthesis methods. Additionally, their DNA entrapment efficiency, cell viability, and functional stability profiles were generally superior. These findings provide new insights for comparing different manufacturing methods to create LPNPs with the desired characteristics for effective and safe gene delivery.

Expanding the horizon of transient CAR T therapeutics using virus-free technology.

The extraordinary success that chimeric antigen receptor (CAR) T cell therapies have shown over the years on fighting hematological malignancies is evidenced by the six FDA-approved products present on the market. CAR T treatments have forever changed the way we understand cellular immunotherapies, as current research in the topic is expanding even outside the field of cancer with very promising results. Until now, virus-based strategies have been used for CAR T cell manufacturing. However, this methodology presents relevant limitations that need to be addressed prior to wide spreading this technology to other pathologies and in order to optimize current cancer treatments. Several approaches are being explored to overcome these challenges such as virus-free alternatives that additionally offer the possibility of developing transient CAR expression or in vivo T cell modification. In this review, we aim to spotlight a pivotal juncture in the history of medicine where a significant change in perspective is occurring. We review the current progress made on viral-based CAR T therapies as well as their limitations and we discuss the future outlook of virus-free CAR T strategies to overcome current challenges and achieve affordable immunotherapies for a wide variety of pathologies, including cancer.

The aryl hydrocarbon receptor (AhR) activation mediates benzo(a)pyrene-induced overexpression of AQP3 and Notch1 in HaCaT cells.

The aim of this study was twofold: (1) evaluate the effect of benzo[a]pyrene (BaP) on expression levels of AQP3 and Notch1 genes in HaCaT cells exposed "in vitro" and (2) investigate the possible biological role of assessed genes by bioinformatics methods. Cells were exposed to increasing concentrations of BaP (0.0-4.0 µM) for 1-4 days. After treatments, cell viability and expression levels of AhR, CYP1A1, AQP3, and Notch1 genes were evaluated. The possible biological role of assessed genes was evaluated using bioinformatics tools. Low cytotoxicity in HaCaT cells dosed with BaP was detected. A significant overexpression (p < .05) of CYP1A1, AQP3, and Notch1 was found in exposed HaCaT cells. The gene expression upregulation was dependent on AhR activation. The bioinformatics analysis showed that these genes were enriched in related cancer signaling pathways. The findings suggest that AQP3 and Notch1 are upregulated by AhR activation in HaCaT cells exposed to BaP.

Expression levels and network analysis of inflammamiRs in peripheral blood mononuclear cells exposed to DDE "in vitro".

Recent studies have demonstrated that dichlorodiphenyldichloroethylene (DDE) induced a pro-inflammatory condition in peripheral blood mononuclear cells (PBMC). However, the molecular mechanisms implicated in this condition are poorly understood. Therefore, this study aimed to evaluate miR-155, miR-126, and miR-21 expression levels in PBMC exposed "in vitro" to DDE. PBMC were dosed with increasing concentrations of DDE (10-80 µg mL-1) at different treatment times (0-24 h). The results showed an up-regulation in the expression levels of assessed miRNAs (miR-155, miR-146, and miR-21) after PBMCs were exposed to DDE. Besides, bioinformatic analysis was performed to understand the biological roles of assessed miRNAs. The bioinformatic analysis shows that assessed miRNAs are associated with regulating signaling pathways involved in cancer, apoptosis, cell cycle, inflammation, metabolism, etc. These findings offer new insights into the molecular mechanisms related to the inflammatory processes and their regulation induced by DDE in PBMC exposed "in vitro".

Highlights of the 'I Congress ecancer Choosing Wisely' March 30 and 31, 2022 Santa Cruz, Bolivia.

The ecancer 'Choosing Wisely' conference was held for the first time in Latin America in Santa Cruz, Bolivia. The event had more than 150 registered attendees in addition to 22 speakers from different countries and different specialities in the field of oncology, who presented topics on prevention, oncological surgery, clinical oncology and palliative care, in order to demonstrate the current evidence of how to approach a patient in daily clinical practice based on the human resources, materials and drugs available, trying to offer the maximum benefit to the patient based on current scientific evidence. In addition to addressing issues of vital importance in breast cancer, during the 2 days of the event, updated information generated in recent years was presented, the results of which will change clinical practice. All the experts were in favour of developing strategies and methods that help us to properly select treatments to optimise resources and reduce the economic toxicity of the most modern and current treatments. This conference was an event of vital importance because it was the first face-to-face event for ecancer and the physicians after difficult years due to COVID-19.

Differential Expression of AhR in Peripheral Mononuclear Cells in Response to Exposure to Polycyclic Aromatic Hydrocarbons in Mexican Women.

The exposure to air pollutants causes significant damage to health, and inefficient cooking and heating practices produce high levels of household air pollution, including a wide range of health-damaging pollutants such as fine particles, carbon monoxide and PAHs. The exposure to PAHs has been associated with the development of neoplastic processes, asthma, genotoxicity, altered neurodevelopment and inflammation. The effects on the induction of proinflammatory cytokines are attributed to the activation of AhR. However, the molecular mechanisms by which the PAHs produce proinflammatory effects are unknown. This study was performed on a group of 41 Mexican women from two rural communities who had stoves inside their houses, used wood as biomass fuel, and, thus, were vulnerable. According to the urinary 1-OHP concentration, the samples were stratified into two groups for determination of the levels of TNF-α, AhR, CYP1B1, miR-125b and miR-155 expression. Our results showed that the CYP1B1, TNF-α, miR-125b and miR-155 expression levels were not statistically different between women with the lowest and highest levels of 1-OHP. Interestingly, high levels of PAHs promoted augmented expression of AhR, which is a protein involved in the modulation of inflammatory pathways in vivo, suggesting that cell signaling of AhR may be implicated in several pathogenesis processes.

Probabilistic human health risk assessment associated with fluoride and arsenic co-occurrence in drinking water from the metropolitan area of San Luis Potosí, Mexico.

A major public health concern in Mexico is the natural contamination of groundwater with fluoride and arsenic. Therefore, this work aimed to evaluate the magnitude of human health risk after determining fluoride and arsenic concentrations in groundwater samples (n = 50) from the Metropolitan area of the city of San Luis Potosi, Mexico. Fluoride levels in water were determined via a potentiometric method using an ion-selective electrode. Arsenic concentrations in water samples were determined with an Atomic Absorption technique. Subsequently, a probabilistic health risk assessment was developed (Monte Carlo Analysis). Fluoride levels in water ranged from 0.20 to 3.50 mg/L. For arsenic, the mean level found in the assessed water samples was 15.5 ± 5.50 µg/L (range: 2.50-30.0 µg/L). In addition, when the probabilistic health risk assessment was completed, a mean HI (cumulative hazardous index) of higher than 1 was detected, indicating a high NCR (non-carcinogenic risk) for children and adults. According to the results found in this study, exposure protection campaigns are imperative in the Metropolitan area of the city of San Luis Potosí, Mexico, to successfully diminish exposure to arsenic and fluoride and, as a consequence, decrease the NCR in the population living in that region of Mexico.

Novel clinical device tracking and tissue event characterization using proximally placed audio signal acquisition and processing.

We propose a new and complementary approach to image guidance for monitoring medical interventional devices (MID) with human tissue interaction and surgery augmentation by acquiring acoustic emission data from the proximal end of the MID outside the patient to extract dynamical characteristics of the interaction between the distal tip and the tissue touched or penetrated by the MID. We conducted phantom based experiments (n = 955) to show dynamic tool/tissue interaction during tissue needle passage (a) and vessel perforation caused by guide wire artery perforation (b). We use time-varying auto-regressive (TV-AR) modelling to characterize the dynamic changes and time-varying maximal energy pole (TV-MEP) to compute subsequent analysis of MID/tissue interaction characterization patterns. Qualitative and quantitative analysis showed that the TV-AR spectrum and the TV-MEP indicated the time instants of the needle path through different phantom objects (a) and clearly showed a perforation versus other generated artefacts (b). We demonstrated that audio signals acquired from the proximal part of an MID could provide valuable additional information to surgeons during minimally invasive procedures.

Human Health Risks Assessment Associated with Polychlorinated Biphenyls (PCBs) in Soil from Different Contaminated Areas of Mexico.

Recent studies have documented environmental contamination by PCBs in soil from different areas in Mexico (industrial, mining, and urban sites). However, the real significance of that soil contamination has not been established. Therefore, the aim of this study was to perform a human health risk assessment (Monte Carlos simulation) to evaluate the probable toxic effects of soils contaminated with PCBs on children in four sites in Mexico. A high non-carcinogenic risk (total nHQ = 1.1E+01; if nHQ ≥1, hazardous health effects cannot be ruled out) was found in Alpuyeca, Morelos, Mexico. Moreover, the total CR (cancer risk) found in Alpuyeca, Morelos is of concern (total CR = 5.1E-03), being that a cut-point of 1.0E-06 has been suggested as a safe level for cancer risk. Taking into consideration the data shown in this research, we conclude that a strategy to protect human health is necessary for the assessed sites.

Urinary trans-trans muconic acid (exposure biomarker to benzene) and hippuric acid (exposure biomarker to toluene) concentrations in Mexican women living in high-risk scenarios of air pollution.

This study aimed to determine t,t-muconic acid (t,t-MA; exposure biomarker for benzene) and hippuric acid (HA; exposure biomarker for toluene) concentrations in the urine of women living in Mexico. In a cross-sectional study, apparently healthy women (n = 104) were voluntarily recruited from localities with a high risk of air pollution; t,t-MA and HA in urine were quantified using a high-performance liquid chromatography (HPLC) technique. Mean urinary levels of t,t-MA ranged from 680 to 1,310 µg/g creatinine. Mean values of HA ranged from 0.38 to 0.87 g/g creatinine. In conclusion, compared to data recently reported in literature, we found high urinary levels of t,t-MA and HA in assessed women participating in this study. We therefore deem the implementation of a strategy aimed at the reduction of exposure as a necessary measure for the evaluated communities.

Adalimumab como alternativa terapéutica en el s¡ndrome de vogt koyanagi harada refractario al infliximab: reporte de caso / Adalimumab, therapeutic alternative in the vogt koyanagi harada syndrome refractory to infliximab: case report

El síndrome de Vogt Koyanagi Harada (VKH) consiste en una panuveitis bilateral que forma parte de los síndromes uveomeníngeos.El tratamiento en estadio crónico es difícil por presentar pobre respuesta a la inmunomodulación, por lo que se recurre a opcionesterapéuticas como agentes biológicos tipo anti-TNF alfa. Se describe el caso de una paciente con VKH severo y resistencia al infliximab,quien mostró respuesta al adalimumab. El adalimumab es un anticuerpo monoclonal humanizado efectivo en casos de resistenciaal infliximab en pacientes con síndrome de VKH crónico persistente. El caso es de interés por ser infrecuente la resistencia a estemedicamento en la práctica clínica, y el uso del activador de plasminógeno tisular contribuyó significativamente en la mejoría visual.

Vogt Koyanagi Harada syndrome (VKH) is a bilateral panuveitis included in the uveomeningeal syndromes. Treatment of its chronicstage is difficult because of poor response to immunomodulation. Other therapeutic options include biological agents such as antiTNFalpha. We present the case of a patient with severe VKH resistant to infliximab that responded to adalimumab. Adalimumab isa humanized monoclonal antibody effective when there is resistance to infliximab in patients with chronic persistent VKH syndrome.The case presented is interesting because of uncommon resistance to this drug in the clinical practice; the use of tissue plasminogenactivator contributed significantly to visual improvement.

DDE and PCB 153 independently induce aryl hydrocarbon receptor (AhR) expression in peripheral blood mononuclear cells.

Recent studies have demonstrated that compounds inducing pro-inflammatory cytokines enhance AhR expression. The aim of this study was 2-fold: (1) to determine if two pro-inflammatory compounds, dichlorodiphenyldichloroethylene (DDE) and 2,2',4,4',5,5'-hexa-chlorobiphenyl (PCB 153), independently affect AhR gene expression in peripheral blood mononuclear cells (PBMC); and (2) if affected, to determine whether the mechanism involved was due to AhR activation or to a pro-inflammatory effect of the chemicals. PBMC isolated from healthy individuals were incubated in the presence of DDE (10 µg/ml) and PCB 153 (20 ng/ml) over time and AhR and CYP1A1 expression was assessed with a real-time PCR technique. The results indicated there was over-expression of the AhR mRNA in PBMC when the cells were treated with DDE and PCB 153. No changes in expression levels of CYP1A1 mRNA were found. Importantly, when the cells were exposed to DDE and PCB 153 in the presence of an antagonist of tumor necrosis factor (TNF)-α, the over-expression of AhR was abolished; as expected, the expression of CYP1A1 was unaffected. In conclusion, these studies demonstrated for the first time an increment of AhR expression "in vitro" in PBMC treated with two pro-inflammatory environmental pollutants, DDE and PCB153. Moreover, the over-expression of AhR was dependent of TNFα induced by DDE and PCB 153 and was independent of AhR activation.

Epigenetic markers of exposure to polycyclic aromatic hydrocarbons in Mexican brickmakers: a pilot study.

A pilot cross-sectional study was carried out in a group of 39 male brick manufacturers in San Luis Potosi, Mexico to identify epigenetic biomarkers of exposure to polycyclic aromatic hydrocarbons (PAHs). A questionnaire was used to compile the smoking and drinking habits, clinical history, working time, and socioeconomic characteristics of the participants. 1-Hydroxypyrene (1-OHP) levels were measured from urine samples using high-performance liquid chromatography, and genomic DNA was isolated from blood samples for methylation analysis using pyrosequencing. The mean 1-OHP level was 0.18 µg g(-1) creatinine (range 0.023-1.11), which was below the expected occupational exposure level. After adjusting for potential confounders, the 1-OHP urine concentration was negatively associated with DNA methylation of the interleukin 12 (ß=-1.57; 95% CI: -2.9 to -0.23; p=0.02) and p53 gene promoters (ß=-2.7; 95% CI: -5.46-0.06; p=0.055). Suggestive negative associations were also found for the TNF-α gene (ß=-3.9; 95% CI:-8.28-0.48; p=0.08) and Alu sequences (ß=-0.55; 95% CI:-1.25-0.16; p=0.12). Although the individual exposures to PAHs as estimated by urinary 1-OHP concentrations were low, changes in specific and global DNA methylation were observed.

Urinary arsenic levels and risk of renal injury in a cross-sectional study in open population.

INTRODUCTION: Arsenic (As) is one of the most ubiquitous elements in nature, and a prolonged exposure has been associated with an increase in the risk of cancer, diabetes mellitus, hypertension and cardiovascular disease. There are few studies addressing the effects of As on albuminuria, tubular injury and biochemical variables as uric acid. AIM. To analyze the association between urinary As levels, albuminuria, and al-microglobulin as marker of tubular injury. MATERIAL AND METHODS: This is a cross-sectional, and comparative study done in 5 communities localized close to Queretaro City. Subjects with no antecedents of renal disease, diabetes, hypertension, or industrial exposure to As were included. A questionnaire about risk factors for arsenic exposure was done, blood was taken for biochemical analysis and a spot urine sample was collected for albumin, alpha1-microglobulin, and As measurements. RESULTS: A total of 90 adult persons were included with no antecedents of renal disease, diabetes or hypertension; the mean age was 40.9 +/- 12.9 years and the median for urinary As levels was 15 microg/gr Cr (range 0.56-89.2 microg/gr Cr), 10 (11.1%) persons had critical levels > 50 microg/gr Cr. Age more than 50 years old [OR 2.48 IC95 (0.9-6.6)] and place of residence were the most important risk factors associated with higher levels of As. There was association between urinary As levels and al-microglobulin urinary excretion (r2 = 0.07, p = 0.01) but not with albuminuria or other biochemical variables. CONCLUSIONS: This is the first study in Mexico to show an association between As and urinary excretion of al-microglobulin as marker of early renal injury. We did not found association with albuminuria or other serum biochemical variables. Arsenic may be considered as a risk factor for tubular injury.

Variability in DDT-induced apoptosis in Mexican indigenous populations.

CONTEXT: In previous studies, we showed that DDT and its metabolites are able to induce apoptosis of peripheral blood mononuclear cells (PBMC) "in vitro" and "in vivo", by a mechanism involving oxidative stress. OBJECTIVE: The objective of this study was to evaluate the mechanism by which DDT induces apoptosis in PBMC in children exposed to DDT and its metabolites. MATERIALS AND METHODS: Eligibility criteria included children who: (1) have lived in the selected community since birth, (2) were between 6 and 14 years of age at the time of the study, (3) had not been exposed to medicaments or tobacco smoke, and (4) had had no infectious diseases in the month prior to the study. DDT and its metabolites were quantified using gas chromatography with an electron capture detector, PBMC apoptosis was measured using the TUNEL assay, DNA damage and oxidative damage were studied using the comet assay. RESULTS: Apoptosis correlated to DDE exposure (p=0.040), as previously found. DNA damage also correlated to DDT (p=0.005) and DDE (p=0.004) levels. However, neither exposure to DDT or DDE and oxidative damage, nor oxidative damage and apoptosis, were significantly correlated. Children living in Lacanja, Chiapas, one of the communities studied in this work, had the highest levels of exposure to DDT and its metabolites, yet had the lowest percentage of apoptosis. CONCLUSION: Resistance to DDE-induced apoptosis was found in children from one community. Further studies are needed in order to understand the mechanism involved in this apoptosis resistance.

Toxicología clínica comunitaria / Community Clinical Toxicology

En algunos países de América Latina las intoxicaciones agudas se manejan de manera profesional por médicos especialistas en la mate-ria. Algo similar ocurre con las intoxicaciones crónicas de origen laboral en el sector formal. No obstante, una realidad diferente ocurre en cuanto a la evaluación de las intoxicaciones crónicas de origen ambiental, dado que éstas por su naturaleza, son más difíciles de diagnosticar. Para el tratamiento de las intoxicaciones agudas se han organizado Centros de Información y Atención Toxicológica, pero para las intoxicaciones crónicas ambientales no se ha generado organismos semejantes. Por consiguiente, en este trabajo sugerimos un modelo de atención de la intoxicaciones crónicas a través de grupos multidisciplinarios bajo el esquema de una nueva disciplina: la Toxicología Clínica Comunitaria, cuyo objetivo sería la atención simultánea de las intoxicaciones agudas que generalmente se atienden en un ámbito hospitalario y de las intoxicaciones ambientales que por lo normal se presentan a nivel comunitario. El objetivo final es aprovechar la experiencia que existe en la Región en cuanto a Toxicología Clínica para organizar el trabajo comunitario.

In some Latin American countries acute intoxication is professionally managed by specialized physicians qualified in the area. Something similar occurs with work-related chronic intoxication in the formal sector. However, a different reality prevails for the assessment of chronic intoxication of environmental origin, since it is by definition more difficult to diagnose. For treatment of acute intoxication, Toxicological Information and Care Centers have been set up, though similar bodies have not been created for chronic environmental intoxication. Therefore, in this study a model of chronic intoxication care is proposed, using multidisciplinary teams adopting a new approach, namely Community Clinical Toxicology, the goal of which would be the simultaneous care of acute intoxication which is generally treated in hospital, and environmental intoxication that is normal dealt with at community level. The ultimate goal is to take advantage of the expertise that exists in the Region in terms of Clinical Toxicology to organize community work.

Revisión de las metodologías sobre evaluación de riesgos en salud para el estudio de comunidades vulnerables en América Latina / Health risk assessment methodologies for the study of vulnerable communities in Latin America / Revisão das metodologias sobre avaliação de riscos na saúde para o estudo de comunidades vulneráveis na América Latina

Los métodos para evaluar el riesgo en salud se basan, en general, en el monitoreo ambiental y en la estimación de la exposición a través de modelos matemáticos. La incertidumbre de tal estrategia es grande. En consecuencia, para incrementar la certidumbre sobre la evaluación de la exposición a los contaminantes, se ha propuesto el empleo de biomarcadores. No obstante, la complejidad de los nuevos escenarios de riesgo obliga a evaluar no solamente a las poblaciones humanas sino también al resto de la biota. Asimismo, factores ambientales, sociales y de salud, al afectar la vulnerabilidad, también deben ser considerados para la caracterización del riesgo. Estos factores de vulnerabilidad pueden evaluarse a través de indicadores. Al final, con los análisis ambientales, el uso de biomarcadores y el manejo de indicadores ambientales, sociales y de salud, puede evaluarse el riesgo de manera integrada (humanos y biota). En esta revisión se presentan las diversas estrategias empleadas por este grupo de trabajo para evaluar el riesgo en sitios contaminados, comunidades marginadas y en áreas afectadas por el cambio global climático.

The most commonly used methods for risk assessment are based on environmental analysis and the use of mathematical models for the estimation of exposure. However, the uncertainty of this approach is high, as the models are based on scenarios that may be not the correct ones. In order to decrease the uncertainty, the use of biomarkers has been proposed. Furthermore, considering the complexity of pollution in some sites, these biomarkers can be used both in humans and biota in order to obtain better information for the definition of risks at those sites. In addition to biomarkers, social, health and environmental indicators have to be applied for risk characterization, as different factors of vulnerability can modify the extent of health risks in some communities. At the end, with environmental monitoring and the use of biomarkers and indicators of vulnerability, health risks in humans and biota (integrated risk assessment) can be assessed in different scenarios. In this paper we present the strategies that our group developed for the study of hazardous waste sites, vulnerable communities and areas impacted by climate change.

Os métodos para avaliar o risco na saúde se baseiam, em geral, no monitoramento ambiental e na estimação da exposição através de modelos matemáticos. A incerteza de tal estratégia é grande. Em consequência, para incrementar a certeza sobre a avaliação da exposição aos contaminantes, tem sido proposta a utilização de biomarcadores. No entanto, a complexidade dos novos cenários de risco obriga a avaliar não somente as populações humanas mas também ao resto da biota. Da mesma forma, fatôres ambientais, sociais e de saúde, ao afetar a vulnerabilidade, também devem ser considerados para a caracterização do risco. Estes fatôres de vulnerabilidade podem avaliar-se através de indicadores. Finalmente, com as análises ambientais, o uso de biomarcadores e o manejo de indicadores ambientais, sociais e de saúde, pode-se avaliar o risco de maneira integrada (humanos e biota). Nesta revisão se apresentam as diversas estratégias empregadas por este grupo de trabalho para avaliar o risco em lugares contaminados, comunidades marginalizadas e em áreas afetadas pela mudança global climática.

DDT-induced oxidative damage in human blood mononuclear cells.

Recent work indicates that DDT and its metabolites induce apoptosis in different cellular types. However, the mechanism by which DDT generates apoptosis has not been elucidated. In this study, our data demonstrate that the apoptosis induction by DDT and its metabolites in human peripheral blood mononuclear cells (PBMC) is preceded by an increase in the levels of reactive oxygen species (ROS). Cells isolated from healthy individuals were incubated for different intervals of time (0-24 h) and in the presence of increasing concentrations of p'p-DDT, p'p-DDE, or p'p-DDD (0-80 microg/ml). The induction of oxidative stress was then determined by flow cytometry, using the compound 2',7'-dichlorofluorescein diacetate. The control level of ROS was 4.46+/-0.96 IFM, for DDT- and DDD-treated cells we obtained a 19.0-fold increment, whereas for DDE, the increment was 25-fold. ROS induction by DDT and DDE was observed after 1 h of incubation, while for DDD such levels began to be detected at 3 h of incubation; a maximum effect on the ROS production for the three compounds was found at 6 h of treatment. A significant level of ROS was induced by DDT, DDE, and DDD only at 60 and 80 microg/ml. Finally, to find an association between generation of ROS and apoptosis induction, cells incubated with DDT, DDE, and DDD were evaluated for apoptosis induction and generation of oxidative stress. Our results show that an increase in ROS was accompanied by apoptosis of PBMC in vitro. Moreover, N-acetyl-L-cysteine significantly inhibits the apoptosis induction.