Giuliana Abbadessa

Sickle Cell Disease is among the top 20 causes of mortality worldwide. Hydroxyurea is one of the most readily available and economical solutions approved for the treatment of Sickle Cell Disease patients: it induces the synthesis of foetal haemoglobin and decreases the inflammatory phenotype. Hydroxyurea shows high interindividual variability, re-quiring individualized dosing to balance therapeutic efficacy against escalating toxicity. In addition to non-genetic factors, hereditary variables can also be investigated to adopt a more tailored dosing regimen according to each patient's demographic, pharmacokinetic and pharmacogenetic profile. This study evaluated the possible influence of different genetic variants on hydroxyurea pharmacokinetics, by conducting a retrospective analysis on 79 treated patients, stratified by ethnicity, age and sex. SLC22A3*698 C>G variant was found to be the most significantly associated with an en-hanced function of the encoded transporter, improving the drug elimination process. ABCB1 1236 C>T, ABCB1 2677 G>A, ABCB1 3435 C>T, CYP2B6 516 G>T, CYP2C19*2 G>A, CYP2C9*3A A>C, CYP2D6*10 A>G, MAP3K5 13257 C>T, MAP3K5 6292 C>A and OCT1 480 C>G gene variants were instead found linked to a general loss-of-function phenotype, reflecting slower and less efficient drug metabolism. These findings demonstrate the importance of assessing hydroxyurea pharmacogenetics while taking into account different ethnic and demographic variables, in order to support a more personalized and effective therapeutic approach for sickle cell disease patients.

Background: Endogenous estradiol/progesterone (E2/P4) regulates neuronal mitochondrial bioenergetics and redox balance. However, the effects of combined oral contraceptive (COC) steroids under physiologically relevant hormonal conditions remain poorly understood. Therefore, this study aims to investigate how COC steroids modulate mitochondrial function within the defined E2/P4 hormonal milieus. Methods: Human SH-SY5Y cells (Cytion) were starved for 48 h in Cytion medium without FBS and with 1% ITS, and then exposed for 48 h to six conditions: vehicle (F0); follicular-like E2/P4 (F1); luteal-like E2/P4 (F2); F1 + dienogest/ethinylestradiol (DNG/EE; F3); F2 + DNG/EE (F4); and DNG/EE alone (F5). The primary endpoints were mitochondrial membrane potential (JC-1 red/green ratio) and ROS (H₂DCFDA/DCF); nitric oxide (DAF-FM) was also recorded. Hoechst 33342 nuclear fluorescence served both as a per-well proxy of cell number and as a proportionality factor for normalization. Results: Follicular- and luteal-like backgrounds were associated with distinct ΔΨm/ROS set-points. The addition of DNG/EE was accompanied by background-dependent shifts, generally characterized by higher DCF signals and lower JC-1 ratios relative to the vehicle, whereas DAF-FM did not reveal statistically robust changes in NO. Hoechst-based normalization preserved these patterns, suggesting that the observed effects likely reflect per-cell functional modulation rather than differences in cell number, although modest, background-dependent variations in proliferation were observed, with DNG/EE associated with a greater increase in ROS under follicular-like conditions and a more pronounced ΔΨm membrane potential in a luteal-like milieu. Conclusions: These findings suggest that neuroendocrine background may influence mitochondrial ΔΨm/ROS states in SH-SY5Y cells, with physiologically inspired E2/P4 milieus potentially shaping baseline conditions onto which COC components exert context-dependent effects at 48 h. These preliminary findings provide a standardized framework for subsequent image-based analyses of mitochondrial function.

Background: Endogenous estradiol/progesterone (E2/P4) shape neuronal mitochondrial bioenergetics and redox. The mechanisms by which combined oral contraceptive (COC) steroids operate within physiologically relevant hormonal milieus remain poorly defined. Methods: Human SH-SY5Y cells (Cytion) were starved 48 h in Cytion medium without FBS and with 1% ITS, then exposed for 48 h to six conditions: vehicle (F0); follicular-like E2/P4 (F1); luteal-like E2/P4 (F2); F1 + dienogest/ethinylestradiol (DNG/EE; F3); F2 + DNG/EE (F4); and DNG/EE alone (F5). Primary endpoints were mitochondrial membrane potential (JC-1 red/green ratio) and ROS (H₂DCFDA/DCF); nitric oxide (DAF-FM) was also recorded. Hoechst 33342 nuclear fluorescence served both as a per-well proxy of cell number and as a proportionality factor for normalization. Results: Follicular- and luteal-like backgrounds produced distinct ΔΨm/ROS set-points. Superimposing DNG/EE generated background-dependent shifts marked by higher DCF and lower JC-1 ratios versus vehicle; DAF-FM indicated concordant NO changes. Hoechst-based normalization preserved these patterns, indicating per-cell functional modulation rather than cell-number artifacts; proliferation differences were modest and background-dependent. Conclusions: Neuroendocrine background matters: physiologic E2/P4 milieus prime mitochondrial ΔΨm/ROS/NO states in SH-SY5Y, onto which COC components impose con-text-dependent modulation at 48 h. These preliminary findings provide a standardized framework for subsequent image-based mitochondrial phenotyping.

Recurrent aphthous stomatitis (RAS) is the most common ulcerative disorder of the oral cavity, although its etiology is still unknown. The present study aimed to identify the proteomic profile associated with the RAS inflammatory process, thereby enhancing our understanding of its etiopathogenesis. We compared salivary protein profiles of RAS patients during an active episode of oral ulceration (30 patients, mean age 36.9) to those from healthy donors without a history of RAS (30 healthy subjects, mean age 37.9). Using 2D-electrophoresis and mass spectrometry (MALDI-TOF) analysis, we identified 17 proteins that were differentially expressed in the two groups. Notably, Cystatin SN (CST1) appeared to be significantly downregulated in RAS patients. These findings were validated by Western blot analysis: CST1 was detected in only 3 of the 30 RAS cases, while it was strongly expressed in all the healthy subjects. Although preliminary, our results suggest a potential role for CST1 in the etiopathogenesis of RAS. Interestingly, the relative absence of CST1 in RAS patients seems to align with some clinical and molecular features of this disease.

Background: Hydroxyurea is approved for the treatment of paediatric and adult sickle cell disease patients. It causes the synthesis of foetal haemoglobin and decreases platelets and granulocytes, but with a high interindividual variability, requiring higher dosages and escalating toxicity. Hereditary variables should be investigated to personalise treatment. We evaluated the possible influences of OCT1 and MAP3K5 gene polymorphisms on hydroxyurea pharmacokinetics. Methods: We conducted a retrospective analysis on 79 treated patients. The polymorphisms of OCT1 (rs683369 G > C) and MAP3K5 (rs9376230 C > A and rs9483947 C > T) were genotyped. Results: Sub-Saharan patients with the OCT1 rs683369 GG genotype showed a lower drug half-life, compared to those with the GC genotype. In sub-Saharan paediatric female patients, the OCT1 rs683369 GG genotype was associated with a lower t1/2 than the GC genotype. Conclusions: The findings demonstrate for the first time how crucial it is to assess the pharmacogenetics of hydroxyurea by taking into account the two sexes in different groups. Additionally, the data were evaluated with consideration for ethnic groups and individually for adults and children. Pharmacogenetic studies could improve the clinical management of hydroxyurea.

Background: Hydroxyurea (HU), also known as hydroxycarbamide, is an oral ribonucleotide reductase inhibitor. In 1999, the United States Food and Drug Administration (FDA) approved HU for the treatment of sickle cell disease (SCD). Since then, it has become the cornerstone in the management of SCD patients, helping to reduce vaso-occlusive crises, acute chest syndrome, the need for blood transfusions, hospitalizations and mortality. There is considerable variability among individuals in HU pharmacokinetic (Pk) parameters that can influence treatment efficacy and toxicity. The objective of this work is part of a clinical study aimed at investigating HU Pk and determining the optimal sampling time to estimate the Area Under the Curve (AUC) in SCD patients. Methods: HU plasma concentration in 80 patients at various time points (2, 4, 6, 24 h) following a 48-h drug washout was quantified using High-Pressure Liquid Chromatography (HPLC) coupled with an ultraviolet (UV) detection method previously described in the literature and adapted to new conditions with partial modifications. Results: The mean HU administered dose was 19.5 ± 5.1 mg/kg (range: 7.7–37.5 mg/kg). The median AUC quantified in plasma patients was 101.3 mg/L/h (Interquartile Range (IQR): 72.5–355.9) and it was not influenced by the weight-based dose. However, there was a strong positive correlation between AUC and Body Mass Index (BMI) as well as dose per Body Surface Area (BSA). Along with a three-point approach for AUC determination present in the literature, we show results obtained from a four-point sampling strategy, which is more useful and effective for better optimizing dose escalation to the maximum tolerated dose (MTD). Moreover, we observed that most patients achieved the maximum HU plasma concentration two hours after drug administration, regardless of age differences. Conclusions: HU treatment, which represents a milestone in the treatment of SCD due to its ability to reduce disease complications and improve patients’ quality of life, requires careful monitoring to optimize the individual dose for saving potential side effects and/or adverse events.

Sleep is a fundamental human need; sleep disruption, in fact, causes an increase in the activity of the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis, metabolic effects, changes in circadian rhythms, and pro-inflammatory responses. The scientific literature is finally starting to pay attention to the central role of sleep alterations in patients health. Oxaliplatin is extensively used for the treatment of gastrointestinal cancer and other malignancies, with an increased frequency of use in recent years. This study aims to understand the effects of sleep complaints on health and quality of life in cancer patients treated with oxaliplatin. A study has been conducted through the creation and distribution of questionnaires to patients to investigate their complaints about sleep quality. We observed significant differences between males and females in evaluating sleep hygiene scores, the Pittsburgh Sleep Quality Index, and previous difficulty sleeping. Moreover, in females, stress, worries, and anxiety seem to play a negative role in the sleep hygiene score. The obtained results could improve the interest of healthcare personnel and caregivers in sleep quality in patients undergoing chemotherapy.

Sleep is a fundamental human need: sleep disruption, in fact, causes an increase in the activity of the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis, metabolic effects, changes in circadian rhythms and pro-inflammatory responses. Sci-entific literature is finally starting to pay attention to the central role of sleep-wake rhythm alterations for patients health. Oxaliplatin is used extensively for the treatment of gastrointestinal cancer and other malignancies, with increased frequency of use in recent years. This study aims to understand the effects of sleep-wake rhythm alterations on health and quality of life on cancer patients treated with oxaliplatin. Study has been conducted through the creation and distribution of questionnaires to patients to inves-tigate the potential alteration of the sleep-wake quality. Evaluating the influence of the collected variables on sleep quality, only liver and biliary tract and pancreas tumor lo-calization, in males, and previous difficulty sleeping, and anxiety, worries and stress, in females, resulted statistically significant. The results obtained may serve as foundation for designing awareness campaigns and updating of patients, healthcare personnel and caregivers, to improve health and quality of patients life.

Background and Objectives: Sexual violence (SV) is a major global public health concern. While socioeconomic factors and familial relationships have been widely reported to contribute to SV, the role of alcohol consumption should not be ignored. Indeed, alcohol can impair cognition, distort reality, increase aggression, and ease drug-facilitated sexual assault. This retrospective study aims to explore the relationship between alcohol consumption and SV by examining the prevalence, characteristics, and consequences of violence episodes. Materials and Methods: A total of 1481 women accessed the Rape Centre “Centro Soccorso Violenza Sessuale” in Turin, Italy between 2008 and 2019, with 223 reporting alcohol consumption before the assault. Results: The alcohol group had a younger age profile, predominantly within the 18–25-year-old category. SV incidents involving alcohol consumers were more likely to occur in public places or in someone else’s home, while the non-alcohol-consuming group experienced more violence in their own homes. Acquaintances and unknown individuals were primarily responsible, whereas partners were the most common perpetrators of violence against non-alcohol-consuming women. Alcohol consumers sought medical attention sooner after the assault and exhibited more symptoms and injuries, particularly of neurological origin. Concurrent use of recreational drugs was higher among alcohol consumers. The logistic regression analysis revealed higher odds of injury for Italian women and those in the 18–35 age groups after consuming alcohol. Conclusions: This study contributes to the understanding of the relationship between alcohol consumption and SV. The prevalence of alcohol-related sexual aggression is lower compared to that shown in previous studies. Nationality, age, and assailant identity influence SV dynamics. These findings can guide well-targeted interventions and prevention strategies to address SV and inform communities facing similar challenges.

Orotic acid (OA) is an intermediate metabolite of pyrimidine nucleotide biosynthesis and represents a minor diet constituent. The measurement of urinary orotic acid is useful in confirming the diagnosis of hereditary metabolic diseases. Moreover, it could be of interest to know how the physiological concentration of this metabolite changes in relation to different conditions of clinical normality. The purpose of this study was to determine the orotic acid concentration in the urine of healthy patients, to observe normal oroticuria and to evaluate if the expression of pyrimidine intermediate biosynthesis differs between healthy males and females. The orotic acid concentration in urine was performed via the ICH M10-validated analytical method. Unexpectedly, females showed a greater oroticuria than males in pediatric age (0–10); conversely, we did not find significant differences until 70 years of age. The LC-MS/MS method was suitable for use in the differential diagnosis of hereditary metabolic disease and metabolic monitoring of anticancer drug-induced toxicity. The analytical protocol was found to be rapid and ideal, and was used in the routine analysis of a clinical chemistry laboratory. The biochemical aspects related to the expression of pyrimidine biosynthesis should be further investigated in light of the obtained results.