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Background: Acne vulgaris is a common skin condition that plagues many individuals and reflects on their quality of life. The pathogenesis of acne is multifactorial and is subject of continuous investigation and research. Infection, inflammation, and oxidative stress are among the numerous theories explaining the pathogenesis of acne. Meanwhile, the etiological factors of acne vulgaris share common grounds with immune alterations during toxoplasmosis. Objectives: Investigation of a possible association between toxoplasmosis and the pathogenesis of acne vulgaris. Subjects and Methods: Serum anti-Toxoplasma IgG and glutathione (GSH) were measured in 43 acne patients and 40 healthy control subjects. Serological detection of anti-Toxoplasma IgG was performed using enzyme linked immunosorbent assay (ELISA) technique and GSH level in serum was detected using colorimetry. In addition, clinical evaluation of the acne lesions was assessed. Results: GSH was significantly lower in seropositive acne patients as compared to seronegative ones. No difference in seroprevalence and anti-Toxoplasma IgG titre was observed between cases and controls. Clinical evaluation of cases revealed that patients with severe acne showed a higher Cardiff acne disability index (CADI) score. Conclusion: The reduction of GSH in seropositive cases suggests a contributing role of toxoplasmosis to the development of acne by altering the oxidative balance. The involvement of toxoplasmosis in the pathogenesis of multi-factorial clinical conditions such as acne vulgaris should be focused upon since latent infection is widely present among the Egyptian population. Toxoplasmosis and acne vulgaris Hamed et al., 159 The current study aims at investigating the existence of this association by measuring serum levels of GSH in acne vulgaris patients, with serological evidence for the presence or absence of toxoplasmosis. SUBJECTS AND METHODS Study subjects and setting: The current study is a case-control analytical study, in which venous blood samples were collected from 43 acne vulgaris patients (age range 15-55 years) attending the Kasr Al Ainy Dermatology Outpatient Clinic, Cairo University, Egypt. Forty healthy individuals not suffering from acne were considered as control subjects. Inclusion criteria include both male and female cooperative patients. Exclusion criteria include patients suffering from disease conditions related to toxoplasmosis other than acne or having been previously diagnosed as suffering from toxoplasmosis, or having previously received anti-Toxoplasma therapy, or badly collected samples. Study parameters: Data collection was performed by a specialized dermatologist covering socio-demographic and clinical data. Serological anti-Toxoplasma IgG was detected using ELISA[7], while GSH level in serum was detected using colorimetry[8]. Specimen collection and preparation: Venous blood samples (5 ml) were drawn under aseptic conditions into sterile blood tubes without additives. Collected blood samples were centrifuged at 3,000 rpm for 5 min for sera preparation and separation. Prepared serum samples were coded and stored at -20oC until tested. Clinical evaluation of patients: In addition to the socio-demographic and clinical data, severity scaling using the global acne severity score (GEA Scale)[9], as well as the assessment of the quality of life using the CADI were performed[10]. The GEA scale depends on the assessment of the dominant lesions, the degree of inflammation and the area of skin involvement. The CADI is a 5-item questionnaire concerned with evaluating the emotional and social impact of acne lesions on the quality of life of patients. Its score ranges from 0-15. Also, the skin type of patients was evaluated in accordance to the Fitzpatrick skin type classification[11]. This grading system classifies the skin type from 1-6 according to the degree of melanin pigmentation in the skin where the fairest skin is designed as type 1, while the dark brown skin is designed as type 6. ELISA for the detection of anti-Toxoplasma IgG in serum[7]: The assay was performed and evaluated according to the manufacturer’s instructions provided in the Toxoplasma IgG ELISA CALBIOTECH test kit supplied by Beta Lab, USA. Diluted patients’ sera were added to wells coated with purified Toxoplasma antigen. Anti-Toxoplasma IgG specific antibody, if present, bounded to the antigen. All unbound material was washed away, and the enzyme conjugate was added to bind to the antibodyantigen complex, if present. Excess enzyme conjugate was washed off and a substrate was added. The intensity of the colour generated was proportional to the amount of IgG antibody in the sample. Reading the absorbance of each sample on an ELISA reader was done at a wavelength of 450 nm. The dual wavelength was measured with a reference filter at 620 nm. Calculation of the sample titre was done according to the following equation: Antibody titre = absorbance of each well/cut off value Cut off value = optical density of the calibrator (1.908) multiplied by the calibrator factor on calibrator bottle (0.35). Results were interpreted as follows: <0.9 = negative; 0.9-1.1 = borderline; < 1.1 = positive. Colorimetric method for estimation of serum GSH[8]: This was performed as described in the reduced GSH kit provided by BIOACTIVIA DIAGNOSTIC, Egypt. The method is based on the reduction of 5,5' dithiobis (2-nitrobenzoic acid) (DTNB) with GSH to produce a yellow compound. The reduced chromogen was directly proportional to the GSH concentration and its absorbance was measured at 405 nm. The calculation of GSH was expressed in mmol/L. Reference value stated in the kit was 4 mmol/L. Statistical analysis: Microsoft excel 2013 was used for data entry and the statistical package for social science (SPSS) version 24 (SPSS, Armonk, New York: International Business Machines Corporation) was used for data analysis. Simple descriptive statistics (arithmetic mean and standard deviation) were used for the summarization of quantitative data and frequencies were used for qualitative data. T-independent and one-way ANOVA were used to compare normally distributed quantitative data. The level of significance was set at probability (P) value <0.05[12]. Ethical considerations: The study was conducted following international guidelines and in accordance with the ethical guidelines of the Faculty of Medicine, Cairo University. Informed consent was obtained from all patients included in this study.

Investigation of the effect of latent toxoplasmosis on oxidative stress in acne vulgaris patients