The Biological and Molecular Action of Ozone and Its Derivatives: State-of-the-Art, Enhanced Scenarios, and Quality Insights.

The ultimate objective of this review is to encourage a multi-disciplinary and integrated methodological approach that, starting from the recognition of some current uncertainties, helps to deepen the molecular bases of ozone treatment effects on human and animal well-being and to optimize their performance in terms of reproducibility of results, quality, and safety. In fact, the common therapeutic treatments are normally documented by healthcare professionals' prescriptions. The same applies to medicinal gases (whose uses are based on their pharmacological effects) that are intended for patients for treatment, diagnostic, or preventive purposes and that have been produced and inspected in accordance with good manufacturing practices and pharmacopoeia monographs. On the contrary, it is the responsibility of healthcare professionals, who thoughtfully choose to use ozone as a medicinal product, to achieve the following objectives: (i) to understand the molecular basis of the mechanism of action; (ii) to adjust the treatment according to the clinical responses obtained in accordance with the principles of precision medicine and personalized therapy; (iii) to ensure all quality standards.

A New Treatment for Local Adiposity with Ascorbic Acid and Ascorbyl-Palmitate Solution: Clinical and Histological Study.

BACKGROUND: Localized adiposity (AL) is the accumulation of subcutaneous adipose tissue, placed in definite anatomic areas, building up an alteration of the body silhouette. The aim of the present clinical and histological study is to assess the effectiveness of an injectable solution containing sodium salt of ascorbic acid 0.24% and surfactant agent at 0.020% ascorbyl-palmitate (SAP) for treating local adiposity. METHODS: Eighty healthy female adult patients were selected, suffering from local adiposity in the abdominal region. The patients underwent a cycle of 6 sessions, with biweekly treatments, without the addition of any active ingredient. Direct infiltration of pharmacologically active SAP solutions into the adipose tissue with a long needle, very similar to the needles used for spinal anesthesia, was performed. This procedure is quick and painless (does not require any anesthesia) with moderate infiltration speed. RESULTS: All the patients treated showed good results with good satisfaction of the circumferential reductions. Before treatment: Waist (cm) 78.8 ± 10.6 and hip 93.6 ± 9.0 with WHR 0.84 ± 0.07. After treatment: Waist (cm) 70.8 ± 9.6 and hip 92.6 ± 8.0 with WHR 0.76 ± 0.06. Indeed, signs of adipocyte apoptosis were observed in subcutaneous skin after injection of SAP. CONCLUSION: The results showed in the present study suggest that the SAP utilized induces apoptosis of adipocytes and could be of use as a safe and effective method with which to eliminate subcutaneous abdominal fat. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Proteomic and Ultrastructural Analysis of Cellulite-New Findings on an Old Topic.

: Background: Cellulite is a condition in which the skin has a dimpled lumpy appearance. The main causes of cellulite development, studied until now, comprehends modified sensitivity to estrogens, the damage of microvasculature present among dermis and hypodermis. The differences of adipose tissue architecture between male and female might make female more susceptible to cellulite. Adipose tissue is seen to be deeply modified during cellulite development. Our study tried to understand the overall features within and surrounding cellulite to apply the best therapeutic approach. METHODS: Samples of gluteal femoral area were collected from cadavers and women who had undergone surgical treatment to remove orange peel characteristics on the skin. Samples from cadavers were employed for an accurate study of cellulite using magnetic resonance imaging at 7 Tesla and for light microscopy. Specimens from patients were employed for the proteomic analysis, which was performed using high resolution mass spectroscopy (MS). Stromal vascular fraction (SVF) was obtained from the samples, which was studied using MS and flow cytometry. RESULTS: light and electron microscopy of the cellulite affected area showed a morphology completely different from the other usual adipose depots. In cellulite affected tissues, sweat glands associated with adipocytes were found. In particular, there were vesicles in the extracellular matrix, indicating a crosstalk between the two different components. Proteomic analysis showed that adipose tissue affected by cellulite is characterized by high degree of oxidative stress and by remodeling phenomena. CONCLUSIONS: The novel aspects of this study are the peculiar morphology of adipose tissue affected by cellulite, which could influence the surgical procedures finalized to the reduction of dimpling, based on the collagen fibers cutting. The second novel aspect is the role played by the mesenchymal stem cells isolated from stromal vascular fraction of adipose tissue affected by cellulite.

Prognostic relevance of oxidative stress measurement in chronic lymphocytic leukaemia.

OBJECTIVE: To evaluate the prognostic significance of oxidative stress (OS) and antioxidant defence status measurement in patients with chronic lymphocytic leukaemia (CLL). METHODS: d-ROMs test and BAP test were evaluated at diagnosis of 165 patients with CLL and correlated with clinical-biological features and prognosis. RESULTS: An increased oxidative damage (d-ROMs test) and a reduced antioxidant potential (BAP test) were found in CLL patients than normal controls (P<.0001). CLL patients with higher d-ROMs values had higher number of circulating white blood cells and lymphocytes, and higher values of ß2 -microglobulin. Higher d-ROMs values were found in female (P=.0003), in patients with unmutated IgVH (P=.04), unfavourable cytogenetics (P=.002) and more advanced clinical stage (P=.002). Higher BAP test values were found in patients expressing CD49d (P=.01) and with more advanced clinical stage (P=.004). At a median follow-up of 48 months, CLL patients with d-ROMs ≥418 CARR U were found to have a shorter time to first treatment (TFT) (P=.0002), and a reduced survival (P=.006) than others. CLL patients with BAP test values ≥2155 µmol/L had a shorter TFT (P=.008) and a shorter survival (P=.003). CONCLUSIONS: OS can affect CLL patients by concomitantly increasing reactive oxygen metabolites production and decreasing antioxidant defences.

Identification of oxidative stress biomarkers in apical periodontitis: A scoping review with bibliometric analysis.

Oxidative stress (OS) is a ubiquitous process for protecting against microorganisms' challenges. This review maps the most used methods for obtaining samples and analysing reactive oxygen species levels in apical periodontitis, following the PRISMA Extension for Scoping Reviews and is registered in Open Science Framework ([https://doi.org/10.17605/OSF.IO/D5U76]). A systematic search was conducted in electronic databases MEDLINE (PubMed), Embase, Scopus, Web of Science, LILACS, SciElo, OATD and DANS up to 17 July 2023. A total of 18 studies were included, with periapical tissue being the most common sample. Twenty-eight different oxidative stress markers were identified, with inducible nitric oxide synthase being the most prevalent. The use of diverse biomarkers for oxidative stress assessment lacks specificity in identifying particular OS species for evaluating apical periodontitis and potential systemic effects. Studies are necessary to compare results obtained from less invasive methods (such as saliva and crevicular fluid) with those from periapical lesion samples.

A pilot study on the efficacy of a seaweed mud application in the treatment of cellulite.

BACKGROUND: Cellulite represents a common multi-factorial condition that affects nearly all women and is now recognized as a clinical condition associated with systemic factors and negative psychological effects. Several noninvasive and minimally invasive treatments were developed during the last few years, but limited evidence supports many of them due to lack of evidence, insufficient participants, and potential adverse effects. METHODS: This study aimed to evaluate the efficacy of a seaweed mud application in improving both the structure and function of tissues affected by cellulite. Sixty women with cellulite underwent 4-week applications of seaweed mud on the buttocks and thighs. The following assessments were performed at baseline and after the last treatment: photographic, clinical, and anthropometric evaluation; tests for elasticity and hydration; ultrasonography of cellulite nodules; and cellulite biopsies in the trochanteric region. Patient satisfaction was assessed using a 5-point Likert-scale questionnaire. RESULTS: The treatment resulted in a significant improvement in the severity of cellulite severity between the initial assessment and the 4-week follow-up, with enhanced structure, elasticity, and hydration of the affected tissues. Microscopic analysis of the cellulite biopsies revealed a significant restoration of dermal organization with induced collagen synthesis and reduced inflammation, edema, and lipid deposition following the 4-week seaweed mud applications. Additionally, the treatment led to a remarkable improvement in comfort and satisfaction as well as a reduction in body circumferences. CONCLUSIONS: The cosmetic application of seaweed mud has proven to be a safe, non-invasive treatment for improving the tissue alterations characteristic of cellulite.

Gluteal femoral subcutaneous and dermal adipose tissue in female.

BACKGROUND: During the sexual maturation, gluteal femoral adipose tissue is subjected to numerous modifications, not observable in other regions, in particular in women and less in men. Other authors described this region, but they used imaging techniques having lower resolution, than MRI proposed in this study. High resolution imaging techniques might provide important and more detailed information about the anatomy of gluteal femoral region. METHODS: This study has been performed using 7 T-magnetic resonance imaging and ultrastructural analysis in order to provide accurate description of the subcutaneous adipose tissue and dermis of gluteal femoral region. In this study specimens harvested from cadavers and form living patients have been analyzed. RESULTS: The results showed the presence of three layers: superficial, middle, and deep, characterized by different organization of fat lobules. High resolution imaging showed the adipose papilla that originates from dermis and protrude in subcutaneous adipose tissue. Adipose papilla is characterized by a peculiar morphology with a basement, a neck and a head and these elements represent the functional subunits of adipose papilla. Moreover, ultrastructural study evidenced the relationship between adipocytes and sweat glands, regulated by lipid vesicles. CONCLUSIONS: This study provides important information about subcutaneous and dermal fat anatomy of gluteal femoral region, improving the past knowledge, and move toward a better understanding of the cellulite physiopathology.

Sirtuins and redox signaling interplay in neurogenesis, neurodegenerative diseases, and neural cell reprogramming.

Since the discovery of Neural Stem Cells (NSCs) there are still mechanism to be clarified, such as the role of mitochondrial metabolism in the regulation of endogenous adult neurogenesis and its implication in neurodegeneration. Although stem cells require glycolysis to maintain their stemness, they can perform oxidative phosphorylation and it is becoming more and more evident that mitochondria are central players, not only for ATP production but also for neuronal differentiation's steps regulation, through their ability to handle cellular redox state, intracellular signaling, epigenetic state of the cell, as well as the gut microbiota-brain axis, upon dietary influences. In this scenario, the 8-oxoguanine DNA glycosylase (OGG1) repair system would link mitochondrial DNA integrity to the modulation of neural differentiation. On the other side, there is an increasing interest in NSCs generation, from induced pluripotent stem cells, as a clinical model for neurodegenerative diseases (NDs), although this methodology still presents several drawbacks, mainly related to the reprogramming process. Indeed, high levels of reactive oxygen species (ROS), associated with telomere shortening, genomic instability, and defective mitochondrial dynamics, lead to pluripotency limitation and reprogramming efficiency's reduction. Moreover, while a physiological or moderate ROS increase serves as a signaling mechanism, to activate differentiation and suppress self-renewal, excessive oxidative stress is a common feature of NDs and aging. This ROS-dependent regulatory effect might be modulated by newly identified ROS suppressors, including the NAD+-dependent deacetylase enzymes family called Sirtuins (SIRTs). Recently, the importance of subcellular localization of NAD synthesis has been coupled to different roles for NAD in chromatin stability, DNA repair, circadian rhythms, and longevity. SIRTs have been described as involved in the control of both telomere's chromatin state and expression of nuclear gene involved in the regulation of mitochondrial gene expression, as well as in several NDs and aging. SIRTs are ubiquitously expressed in the mammalian brain, where they play important roles. In this review we summarize the current knowledge on how SIRTs-dependent modulation of mitochondrial metabolism could impact on neurogenesis and neurodegeneration, focusing mainly on ROS function and their role in SIRTs-mediated cell reprogramming and telomere protection.

Pilot study of histology aspect of cellulite in seventy patients who differ in BMI and cellulite grading.

BACKGROUND: Cellulite is a topographic alteration of the skin with unknown etiology and is characterized by the presence of a dimpled or puckered aspect, as resembling an orange peel and cottage cheese or as having mattress-like appearance. AIM: The aim of this research was to find the different histological aspect of cellulite in sixty patients. MATERIALS AND METHODS: A total of 60 women, mean age 48.8 ( ± 11.08) were included in the study. Among these, 11 women were in menopause (18.33%). All patients after physical examination, aesthetic, and dermatological evaluation were subjected to five cellulite biopsies with a 2.0 mm diameter and 1.5 mm in length in the trochanteric region affected by cellulite. The descriptive statistics were performed for each study predictors demographic age, height, bmi, waist, belly, hip thigh, and knee. RESULTS: The histological analysis of the stained slides showed five different histological features were present in the most of patient. CONCLUSION: In conclusion, the outcome of this study shows that the histological evidence does not characterize the different states of cellulite, but several different histological aspects were present in the same patient, which effectively eliminates staging and could consider cellulite as a degenerative disease.

The role of hyaluronic acid and amino acid against the aging of the human skin: A clinical and histological study.

BACKGROUND: In esthetic medicine, different techniques have been used against the aging of the human skin especially in the facial area. Hyaluronic acid is used for improving the quantity of water and extracellular matrix molecule. The aim of this study is a clinical and histological evaluation of the effect of low-molecular-weight hyaluronic acid fragments mixed with amino acid (HAAM) on the rejuvenation the face skin treated with intradermal microinjections. METHODS: Twenty women with mean age 45 range from 35 to 64 were studied, thereof 8 in menopause and 12 of childbearing age. The patients were treated with the HAAM products by mesotherapy technique; before and after 3 months of the therapeutic procedure, each patient underwent small biopsies with a circular punch biopsy. RESULTS: The clinical results of the present study showed that the administration of the dermal filler containing fragments of hyaluronic acid between 20 and 38 monomers and amino acid via dermis injection technique produces an esthetic improvement in the faces of the treated patients, while the histological evaluation shows an increased fibroblast activity with the production of type III reticular collagen and increased number of vessels and epidermis thickness. CONCLUSIONS: The clinical and histological assessment showed that subcutaneous HAAM infiltration has a significant impact on the dermis and clinical aspects of the face.

Assessment of living conditions in wild boars by analysis of oxidative stress markers.

This study demonstrated that it is possible to differentiate wild boars living in habitats with different animal densities by the measurement of oxidative stress markers. Therefore, reactive oxygen metabolites, the antioxidant barrier, i.e., the biological antioxidant potential and the antioxidative power (OXY-Adsorbent), as well as cortisol were measured in freely ranging wild boars. In two different areas of a State Forest in the Campania Region (Italy), 42 freely ranging, managed wild boars were captured with a corral trap, and blood samples were collected. The wild boars were divided by age (>1 year old and <1 year old) and sex (male and female). Animals in one area showed significantly higher values of oxidative stress parameters than those living in the other area. The annual boar censuses of areas highlighted a higher population density in the first area than in the second one, and this would explain the results obtained. Since the health or nutritional status of animals is reflected by oxidative stress, wild boars in areas with higher animal densities may live under worse conditions than animals in areas with lower densities which becomes evident by measuring oxidative stress markers. As cortisol in blood is only reflecting a short period, the measurement of oxidative stress level may be a better indicator to evaluate the living conditions of wild boars.

Molecular changes in glaucomatous trabecular meshwork. Correlations with retinal ganglion cell death and novel strategies for neuroprotection.

Glaucoma is a chronic neurodegenerative disease characterized by retinal ganglion cell loss. Although significant advances in ophthalmologic knowledge and practice have been made, some glaucoma mechanisms are not yet understood, therefore, up to now there is no effective treatment able to ensure healing. Indeed, either pharmacological or surgical approaches to this disease aim in lowering intraocular pressure, which is considered the only modifiable risk factor. However, it is well known that several factors and metabolites are equally (if not more) involved in glaucoma. Oxidative stress, for instance, plays a pivotal role in both glaucoma onset and progression because it is responsible for the trabecular meshwork cell damage and, consequently, for intraocular pressure increase as well as for glaucomatous damage cascade. This review at first shows accurately the molecular-derived dysfunctions in antioxidant system and in mitochondria homeostasis which due to both oxidative stress and aging, lead to a chronic inflammation state, the trabecular meshwork damage as well as the glaucoma neurodegeneration. Therefore, the main molecular events triggered by oxidative stress up to the proapoptotic signals that promote the ganglion cell death have been highlighted. The second part of this review, instead, describes some of neuroprotective agents such as polyphenols or polyunsaturated fatty acids as possible therapeutic source against the propagation of glaucomatous damage.

Oxidative imbalance in HIV-1 infected patients treated with antiretroviral therapy.

It is generally accepted that oxidative stress is involved in HIV infection. However, the role in oxidative balance of Highly Active Antiretroviral Therapy (HAART) is still debated. In our study we assessed serum oxidant and antioxidant levels in an HIV-1-infected population treated with HAART, and compared them with those of untreated HIV-1 patients and HIV-1-negative subjects. The study included 116 HIV-1-infected patients (86 HAART-treated and 30 untreated), and 46 HIV-negative controls. Serum oxidant levels were significantly higher in the HIV-1 treated group as compared to untreated and control groups. In addition, a decrease of serum total antioxidant status was observed in the HIV-1 treated group. To be noted is that patients who rigorously follow antiretroviral therapy (optimal HAART adherence) have significantly higher oxidative status than those who do not closely follow the therapy (poor HAART adherence). Analysis of variance revealed no significant further increase in oxidative status in HIV-1-infected patients taking antiretroviral and other drugs with the exception of psychiatric drugs (e.g. anxiolytics or antidepressants). Taken together, our results indicate that HAART may affect oxidative stress in HIV-1-infected patients and suggest that antiretroviral therapy plays an important role in the synergy of HIV infection and oxidative stress.

Effects of mode of delivery on pro-oxidant/antioxidant balance in fetal circulation.

Objective: This study was undertaken to assess the influence of mode of delivery on the balance between pro-oxidant/antioxidant systems in fetal circulation. Materials and methods: Both umbilical arterial and venous blood samples were obtained from 37 pregnant women who delivered by spontaneous vaginal delivery (VD group) and from 29 pregnant women who delivered by elective cesarean section (CS group). Oxidative stress and antioxidant activity were evaluated by reactive oxygen metabolites (d-ROMs) and biological antioxidant potential (BAP), respectively. Results: The d-ROMs values of the VD group were higher than that of the CS group in both umbilical arterial and venous blood and these differences were found to be statistically significant (p < .01 and p < .01, respectively). The BAP values of only the umbilical arterial blood were found to be statistically significant (p < .01), with values from the VD group being higher than those of the CS group. In all measurements, the d-ROMs values averaged below 120 CARR U and BAP values averaged above 2200 µmol/L. The ratio of BAP/d-ROMs difference was found to be statistically significant (p < .01) only in the umbilical venous blood, with ratios in the VD group being lower than those in the CS group. Conclusions: Our statistical analyses suggest that vaginal delivery has an effect on increasing oxidative stress as a result of the stress of labor and that an elective cesarean section does not impair the mother's oxidative stress status. Furthermore, the high BAP values in all the measurements suggest that neonates just after birth have the ability to cope with oxidative stress. Rationale  In many studies, the diversity of views on the influence of mode of delivery on the redox status of neonates is likely to be caused by the use of different biomarkers to measure either the oxidative stress, the antioxidant activity, or both. Furthermore, incomplete explanation for sampling cord blood in these studies, either arterial, venous blood or both, complicates matters. To solve the above, this study was designed to assess the effects of mode of delivery on both pro-oxidants, via d-ROMs, and antioxidants, via BAP, in both umbilical arterial and venous blood samples obtained just after birth. There are no existing studies of BAP in both umbilical arterial and venous blood to which we can refer. In conclusion, our study suggests that the pro-oxidant/antioxidant balance in neonates just after birth is better than may be expected when compared to the potentials of adults (including pregnant mothers) according to interpretations of BAP/d-ROMs. This can be understood that neonates may have already been endowed with the ability to cope with oxidative stress, as informed by high BAP values in both umbilical arterial and venous blood. Vaginal delivery may have an effect on increasing oxidative stress as a result of the stress of labors (as measured by d-ROMs), and an elective cesarean section, which has better BAP/d-ROMs in umbilical venous blood than that of vaginal delivery, may not impair the mother's oxidative stress status.

Substances of Interest That Support Glaucoma Therapy.

Glaucoma is a multifactorial disease in which pro-apoptotic signals are directed to retinal ganglion cells. During this disease the conventional outflow pathway becomes malfunctioning. Aqueous humour builds up in the anterior chamber, leading to increased intraocular pressure. Both of these events are related to functional impairment. The knowledge of molecular mechanisms allows us to better understand the usefulness of substances that can support anti-glaucoma therapy. The goal of glaucoma therapy is not simply to lower intraocular pressure; it should also be to facilitate the survival of retinal ganglion cells, as these constitute the real target tissue in this disease, in which the visual pathway is progressively compromised. Indeed, an endothelial dysfunction syndrome affecting the endothelial cells of the trabecular meshwork occurs in both normal-tension glaucoma and high-tension glaucoma. Some substances, such as polyunsaturated fatty acids, can counteract the damage due to the molecular mechanisms - whether ischemic, oxidative, inflammatory or other - that underlie the pathogenesis of glaucoma. In this review, we consider some molecules, such as polyphenols, that can contribute, not only theoretically, to neuroprotection but which are also able to counteract the metabolic pathways that lead to glaucomatous damage. Ginkgo biloba extract, for instance, improves the blood supply to peripheral districts, including the optic nerve and retina and exerts a neuro-protective action by inhibiting apoptosis. Polyunsaturated fatty acids can protect the endothelium and polyphenols exert an anti-inflammatory action through the down-regulation of cytokines such as TNF-α and IL-6. All these substances can aid anti-glaucoma therapy by providing metabolic support for the cells involved in glaucomatous injury. Indeed, it is known that the food we eat is able to change our gene expression.

Oxidative stress in chronic lymphocytic leukemia: still a matter of debate.

There is a large body of evidence showing a strong correlation between carcinogenesis of several types of human tumors, including chronic lymphocytic leukemia (CLL), and oxidative stress (OS). The mechanisms by which OS may promote cancer pathogenesis have not been completely deciphered yet and, in CLL, as in other neoplasms, whether OS is a primary cause or simply a downstream effect of the disease is still an open question. It has been demonstrated that, in CLL, OS concomitantly results from increased reactive oxygen species (ROS) production, mainly ascribable to CLL cells mitochondrial activity, and impaired antioxidant defenses. Interestingly, OS evaluation in CLL patients, at diagnosis, seems to have a prognostic significance, thus getting new insights in the biological comprehension of the disease with potential therapeutic implications.