Correlation between gamma glutamyltransferase fractions and bone quality.

Gamma-glutamyltransferase (GGT) has been recently identified as a bone-resorbing factor. The aim of this study was to investigate the association between plasma GGT fractions levels and bone quality. Plasma GGT fractions were analysed by gel-filtration chromatography. Bone quality was established quantitatively by two micro-CT derived microarchitectural parameters: the BV/TV (mineralised bone volume/total volume), and the SMI (structure model index) that describes the rod-like (low resistant) or plate-like (high-resistant) shape of bone trabeculae. We enrolled 93 patients hospitalised for elective total hip replacement (group Arthrosis, n=46) or for proximal femoral fracture (group Fracture, n=47). Patients within the first quartile of BV/TV (Q1, osteoporotic patients, n=6) showed higher levels of b-GGT fraction [median (min-max): 3.37 (1.42­6.81)] compared to patients with normal bone density (fourth quartile Q4, n=10; 1.40 (0.83­4.36); p=0.0393]. Also, according to SMI, b-GGT value was higher in the subgroup with bone fragility [Q1, n=8: 1.36 (0.43­4.36); Q4, n=8: 5.10 (1.4 ­7.60); p=0.0117]. In conclusion, patients characterised by fragile bone structure showed specifically higher levels of plasma b-GGT activity thus suggesting fractional GGT analysis as a possible biomarker in the diagnosis of osteoporosis.

Ozone autohemotherapy induces long-term cerebral metabolic changes in multiple sclerosis patients.

Ozone autohemotherapy is an emerging therapeutic technique that is gaining increasing importance in treating neurological disorders. A validated and standard methodology to assess the effect of such therapy on brain metabolism and circulation is however still lacking. We used a near-infrared spectroscopy (NIRS) system to monitor the cerebral metabolism and a transcranial Doppler (TCD) to monitor the blood flow velocity in the middle cerebral arteries. Fifty-four subjects (32 neurological patients and 22 controls) were tested before, during, and after ozone autohemotherapy. We monitored the concentration changes in the level of oxygenated and deoxygenated haemoglobin, and in the level of the Cytochrome-c-oxidase (CYT-c). As a primary endpoint of the work, we showed the changes in the brain metabolism and circulation of the entire population. The concentration of oxygenated haemoglobin increased after the reinjection of the ozoned blood and remained higher than the beginning for another 1.5 hours. The concentration of the deoxygenated haemoglobin decreased during the therapy and the CYT-c concentration markedly increased about 1 hour after the reinjection. No significant changes were observed on the blood flow velocity. As secondary endpoint, we compared the NIRS metabolic pattern of 20 remitting-relapsing multiple sclerosis (MS) patients against 20 controls. We showed that by using only 7 NIRS variables it was possible to characterize the metabolic brain pattern of the two groups of subjects. The MS subjects showed a marked increase of the CYT-c activity and concentration about 40 minutes after the end of the autohemotherapy, possibly revealing a reduction of the chronic oxidative stress level typical of MS sufferers. From a technical point of view, this preliminary study showed that NIRS could be useful to show the effects of ozone autohemotherapy at cerebral level, in a long-term monitoring. The clinical result of this study is the quantitative measurement of the CYT-c level changes in MS induced by ozone autohemotherapy.

Oxygen-ozone autohemotherapy in breast cancer patients suffering from fatigue and musculoskeletal pain upon aromatase inhibitors treatment: a case-series study.

OBJECTIVE: In patients with breast cancer and positive hormone receptors, aromatase inhibitors are effective in reducing the risk of recurrences and are active in progressing the disease in this setting. On the other hand, fatigue and painful musculoskeletal side effects can significantly reduce treatment compliance. With no further treatment options to control these symptoms, non-pharmaceutical interventions, such as oxygen-ozone therapy, may play a role in managing rheumatologic symptomatology inasmuch. We have previously reported evidence on the effectiveness of oxygen-ozone in the treatment of pain and fatigue in chronic fatigue syndrome and fibromyalgia patients and in oncological patients as well. PATIENTS AND METHODS: In this study, we reported 6 cases of patients (mean age 64 yrs, all Caucasian females) with breast cancer upon treatment with anastrozole (Arimidex®), suffering from musculoskeletal pain, weakness and fatigue, and therefore treated with oxygen-ozone major autohemotherapy according to the Italian Scientific Society of Oxygen Ozone Therapy (SIOOT) protocol. Pain was measured with a 10-item Numerical Rating Scale (NRS) and fatigue with a 7-item Fatigue Scoring Scale (FSS). RESULTS: A reduction of at least 66% of pain (from 9.43 ±0.54 SD to 2.36 ±1.32 SD, p<0.001) and 66.26% of fatigue were obtained for all the cases. Pain and fatigue disappeared within one month from ozone therapy, and a healthy painless state lasted for many months following the oxygen-ozone therapy. CONCLUSIONS: The oxygen-ozone therapy is a sound opportunity for breast cancer patients to reduce anti-aromatase-induced pain, fatigue, and musculoskeletal symptoms.

Lung cancer and COVID-19: problems and perspectives.

COVID-19 pandemic generated concerns about the healthcare of patients with cancer, not simply because of the formidable impact of COVID-19 patients in the public healthcare system, but also due to the overlapping pathognomonic signs of many forms of lung cancer with lung injuries associated with COVID-19. This report tries to shed light on the issue. We evaluated the great concern of people suffering from lung cancer and also infected with SARS-CoV-2 by discussing evidence and data from current literature. Lung cancer in Italy has represented more than 1 case/4 (27%) in the latest ten years and nevertheless, even due to the concurrence of many complex interplays between COVID-19 and cancer even at the immune level, a consensus protocol and expert guidelines to diagnose and treat lung cancer upon SARS-CoV-2 infection are yet lacking. New insights and consensus panels should be therefore proposed, even at the simplistic level about if priority must be either given to COVID-19 or to cancer therapy.

[Preliminary study to evaluate the meaning of GGT fractions analysis in a population of workers previously exposed to asbestos]. / Studio preliminare volto all'inquadramento del significato preventive del dosaggio di frazioni di GGT in una popolazione di lavoratori ex-esposti ad Amianto.

The GGT enzyme, considered for years only as a marker of liver disease and alcohol abuse, has now revealed a risk of death for many causes. Through a molecular exclusion chromatography on FPLC system (Fast Protein Liquid Chromatography), it is possible to discriminate four fractions of GGT, defined according to the molecular weight: big-GGT, medium-GGT, small-GGT and free-GGT. The objective was to study the preventing meaning of GGT fractions for asbestos-related diseases. This study was conducted on 129 workers previously exposed to asbestos, 22 patients affected by Malignant Pleural Mesothelioma and 107 healthy workers. Our data demonstrated a statistical significant correlation between the fraction free-GGT with the presence of MPM, suggesting a possible role for this molecule as a biomarker for MPM diagnosis. However, being a preliminary study, further studies are warranted to confirm our results.

Fibromyalgia treated with oxygen-ozone auto-haemotherapy (O2-O3-AHT): a case study on 200 patients with a modified 10-PI-NRS evaluation.

OBJECTIVE: Fibromyalgia (FM) is a concerning chronic disease showing as widespread pain, muscle stiffness, sleep disturbances, chronic fatigue, and depressed mood, for which no sound therapy is yet available to date. In this article we assessed a wider patients' cohort the efficacy of oxygen-ozone autohemotherapy (O2-O3-AHT) previously reported. PATIENTS AND METHODS: A number of 200 FM-patients accessed the study and were treated with 3-4 runs of O2-O3-AHT, following their signed consent. A modified 10 points-PI-NRS were used to evaluate pain intensity before and after the conclusion of the complete ozone-treatment cycle (1 month). Kruskall-Wallis' test and other statistics were used at p<0.05 level of significance. RESULTS: A quite complete rehabilitation of the musculoskeletal function and of the overall arthralgia was observed in 76% of the patients at one month of follow-up. The number of patients having a reduction in the PI-NRS score from 10 (the maximal observed) to 3 (including 10-1 and 10-2) following only two runs of O2-O3-AHT was 23.5%, whereas only 17.5% did not show any significant improvement (ΔPI-NRS = 0 or 1), so assessing that the efficacy of O2-O3-AHT, independently from ages, encompassed at least 41% in a moderate way and 64.5% of the treated patients, as a whole. CONCLUSIONS: Oxygen-ozone autohemotherapy represents a formidable therapeutic approach for FM, deserving further studies to be made in order to fully elucidate ozone effect of this pathology.

Sjögren syndrome successfully treated with oxygen-ozone auto-hemotherapy (O2-O3-AHT). a case report.

OBJECTIVE: Sjögren syndrome (SS) is an autoimmune disorder, affecting about 16,000 individuals in Italy, yet lacking a standardized therapy protocol and a plain inclusion in the reimbursed healthcare services. This raises many controversial issues about how managing the SS patient, to relief pain and discomfort and improve patients' health and social life. The ozone therapy resulted successful in previous reports, and therefore, it was used in this case report. CASE PRESENTATION: A 69-years old female outpatient, showing positivity to Schirmer's test, was previously diagnosed as a primary Sjögren syndrome, who later developed an autoimmune thyroiditis and showed the presence of rheumatoid factors. The patient suffered from a marked ocular dryness, subsequently to a purported endothelitis, alongside with fatigue and pain. Laboratory tests showed a positive ANA 1:320 in a speckled pattern with negative anti-SSA and anti-SSB tests. From December 2020 to January 2021 she underwent 2 routes of three sessions of oxygen-ozone autohemotherapy (O2-O3 AHT), as described below and improved, with only 2 sessions, her symptomatology and clinical outcome, as ocular dryness, fatigue and pain, rapidly disappeared. CONCLUSIONS: The use of ozone in the therapy of SS is a straightforward, affordable and feasible approach to treat primary Sjögren syndrome without significant side effects.

Fatigue in post-acute sequelae of SARS-CoV2 (PASC) treated with oxygen-ozone autohemotherapy - preliminary results on 100 patients.

OBJECTIVE: Post-acute sequelae of SARS-CoV2 infection (PASC) are a novel terminology used to describe post-COVID persistent symptoms, mimicking somehow the previously described chronic fatigue syndrome (CFS). In this manuscript, we evaluated a therapeutical approach to address PASC-derived fatigue in a cohort of past-COVID-19 positive patients. PATIENTS AND METHODS: A number of 100 patients, previously diagnosed as COVID-19 positive subjects and meeting our eligibility criteria, was diagnosed having PASC-related fatigue. They were recruited in the study and treated with oxygen-ozone autohemotherapy (O2-O3-AHT), according to the SIOOT protocol. Patients' response to O2-O3-AHT and changes in fatigue were measured with the 7-scoring Fatigue Severity Scale (FSS), according to previously published protocols. RESULTS: Statistics assessed that the effects of O2-O3-AHT on fatigue reduced PASC symptoms by 67%, as a mean, in all the investigated cohort of patients (H = 148.4786 p < 0.0001) (Figure 1). Patients following O2-O3-AHT therapy, quite completely recovered for PASC-associated fatigue, a quote amounting to about two fifths (around 40%) of the whole cohort undergoing ozone treatment and despite most of patients were female subjects, the effect was not influenced by sex distribution (H = 0.7353, p = 0.39117). CONCLUSIONS: Ozone therapy is able to recover normal functionality and to relief pain and discomfort in the form of PASC-associated fatigue in at least 67% of patients suffering from post-COVID sequelae, aside from sex and age distribution.

Ozone Exerts Cytoprotective and Anti-Inflammatory Effects in Cardiomyocytes and Skin Fibroblasts after Incubation with Doxorubicin.

INTRODUCTION: Skin reactions and cardiotoxicity are one of the most common side effects of doxorubicin in cancer patients. The main mechanisms based on the etiopathogenesis of these reactions are mediated by the overproduction of proinflammatory cytokines, metalloproteases, and the disruption of mitochondrial homeostasis. Ozone therapy demonstrated anti-inflammatory effects in several preclinical and clinical studies. The aim of this research is based on the evaluation of cardioprotective and dermatoprotective effects of ozone during incubation with doxorubicin, giving preliminary evidences for further studies in the field of cardio-oncology. METHODS: Human skin fibroblast cells and human fetal cardiomyocytes were exposed to doxorubicin at subclinical concentration (100 nM) alone or combined with ozone concentrated from 10 up to 50 µg/mL. Cell viability and multiple anti-inflammatory studies were performed in both cell lines, with particular attention on the quantification of interleukins, leukotriene B4, NF-κB, and Nrf2 expressions during treatments. RESULTS: Ozone decreased significantly the cytotoxicity of doxorubicin in skin fibroblasts and cardiomyocytes after 24 h of incubation. The best cytoprotective effect of ozone was reached to 30 µg/mL with a plateau phase at higher concentration. Ozone also demonstrated anti-inflammatory effects decreasing significantly the interleukins and proinflammatory mediators in both cells. CONCLUSION: Ozone exerts cardioprotective and dermatoprotective effects during incubation with doxorubicin, and the involved mechanisms are mediated by its anti-inflammatory effects. The overall picture described herein is a pilot study for preclinical studies in oncology.

Antioxidant capacity and protein oxidation in cerebrospinal fluid of amyotrophic lateral sclerosis.

BACKGROUND: The causes of Amyotrophic Lateral Sclerosis (ALS) are unknown. A bulk of evidence supports the hypothesis that oxidative stress and mitochondrial dysfunction can be implicated in ALS pathogenesis. METHODS =: We assessed, in cerebrospinal fluid (CSF) and in plasma of 49 ALS patients and 8 controls, the amount of oxidized proteins (AOPP, advanced oxidation protein products), the total antioxidant capacity (FRA, the ferric reducing ability), and, in CSF, two oxidation products, the 4-hydroxynonenal and the sum of nitrites plus nitrates. RESULTS: The FRA was decreased (p = 0.003) in CSF, and AOPP were increased in both CSF (p = 0.0039) and plasma (p = 0.001) of ALS patients. The content of AOPP was differently represented in CSF of ALS clinical subsets, resulting in increase in the common and pseudopolyneuropathic forms (p < 0.001) and nearly undetectable in the bulbar form, as in controls. The sum of nitrites plus nitrates and 4-hydroxynonenal were unchanged in ALS patients compared with controls. CONCLUSION: Our results, while confirming the occurrence of oxidative stress in ALS, indicate how its effects can be stratified and therefore implicated differently in the pathogenesis of different clinical forms of ALS.

On the incorporation of the non-steroidal anti-inflammatory naproxen into cationic O/W microemulsions.

Microemulsions (ME) containing hexadecyltrimethylammonium bromide (HTAB)/ethanol as surfactant, isopropylmyristate (IM) or butylstearate (BS) as oil phase and aqueous buffer were studied. Pseudo-ternary phase diagrams of the investigated systems were obtained at constant surfactant/cosurfactant molar ratio (1:5) by titration in order to characterize the proportions between the components to obtain clear systems. Oil in water microemulsions were prepared in a wide range of phase volume (phi). UV-vis absorption spectra of naproxen at pH 5.5 showed that the solubility of Np increases significantly in the presence of O/W ME in high phase volumes. For both, IM and BS microemulsions, the dynamic light scattering experiments showed that the size of the oil droplets remains constant in low values of phi, increasing abruptly in high phi values. Phase solubility study revealed that for both IM and BS microemulsions, the drug incorporation followed a straight-line profile in all range of phi. The data could be analyzed through the phase-separation model and the association constants (K) calculated varied from 27 to 90 M(-1), depending on the pH and on the microemulsion oil phase.

gamma-Glutamyl transpeptidase catalyses the extracellular detoxification of cisplatin in a human cell line derived from the proximal convoluted tubule of the kidney.

Nephrotoxicity is a side-effect and the main factor limiting the clinical use of cisplatin. In vivo, the administration of the cysteine-containing tripeptide glutathione (GSH) has been found to reduce nephrotoxicity, but the biochemical mechanism of this protective action is not fully understood. The present study was designed to gain insights into the mechanism by which GSH prevents cisplatin nephrotoxicity. We also wanted to verify the hypothesis of whether the protective action of GSH is mediated by products of the extracellular breakdown of GSH catalysed by gamma-glutamyl transpeptidase (GGT), an enzyme that is highly expressed in kidney tubular cells. The study was performed in HK-2 cells, derived from the immortalisation of human kidney proximal tubule cells. We investigated the influence of modulators of GGT activity and/or thiols on the antiproliferative activity of cisplatin and on the intracellular GSH content. We determined the antiproliferative activity of cisplatin, platinum cellular accumulation and DNA platination following precomplexing of the drug with thiols. The antiproliferative effect of cisplatin was minimally affected by the addition of GSH. However, when the antiproliferative assay was performed in the presence of glycyl-glycine (GlyGly), to serve as a transpeptidation acceptor and thus to stimulate GGT-mediated GSH catabolism, cisplatin-induced growth inhibition was largely prevented. This effect was not mediated through an increase of intracellular GSH levels, which were not affected by the GlyGly supplementation. The thiol dipeptide cysteinyl-glycine, i.e. the GSH catabolite generated by GGT activity, showed a higher reactivity against cisplatin in vitro than GSH, as was shown by the more rapid oxidation of its -SH groups. The cisplatin/GSH or cisplatin/cysteinyl-glycine adducts did not display an antiproliferative effect. However, 2 h precomplexing with GSH in the presence of GGT, or directly with the GSH catabolite cysteinyl-glycine, decreased the antiproliferative effect of cisplatin and drug-induced DNA platination to a greater extent than precomplexing with GSH alone. The results of the present study show that, in HK-2 cells, extracellular GSH decreases the antiproliferative effects of cisplatin only upon its hydrolysis by GGT, thereby supporting the hypothesis that the extracellular metabolism of GSH by GGT plays a role in modulating cisplatin nephrotoxicity. A primary role in the protection of HK-2 cells appears to be played by cysteinyl-glycine, the proximal product of the GGT-mediated hydrolysis of GSH, which shows a high reactivity against CDDP resulting in the rapid inactivation of the drug.

Sentinel node biopsy and internal mammary lymphatic mapping in breast cancer.

BACKGROUND: Currently outer quadrant breast tumors are not felt to have significant drainage to the internal mammary nodal chain. We evaluated the incidence of internal mammary sentinel nodes (IMSN) found with lymphoscintigraphy for all breast quadrants. METHODS: Eighty women were prospectively studied by technitium-sulfur colloid injection, nuclear medicine imaging, and intraoperative gamma probe identification of sentinel nodes. IMSN detected were surgically removed. RESULTS: Ten of 80 ( 12%) patients had IMSN identified in addition to axillary sentinel nodes (AXSN). Four tumors were located medially, 6 (60%) were in the outer quadrants. Metastatic disease was present in 3 of 10 women. Patients with positive IMSN also had positive AXSN. CONCLUSIONS: Patients undergoing lymphatic mapping for the management of breast cancer should have radiolabelled tracer and gamma probe sentinel node identification regardless of tumor location in the breast.

Iatrogenic vascular injuries following lumbar disc surgery. Case report and review of the literature.

Iatrogenic vascular injuries are unusual complications of lumbar disc surgery. The incidence of such injuries is very low but probably underestimated because clinical manifestations may be extremely variable depending on the extension of trauma whether artery or vein are injured or both. Diagnosis is suspected when early signs of retroperitoneal hemorrhage appear, but may often be delayed for weeks or years because formation of a pseudoaneurysm or an arteriovenous fistula which may be of gradual onset and produce only a few symptoms at the onset. Prompt diagnosis and aggressive treatment can hope to improve current mortality of more than 50%. A case of venous and arterial injury is reported, successfully operated on with an original technique of ilio-caval patching to avoid caval ligation. A review of the literature shows the predisposing factors and complex anatomy of such injuries, the clinical picture and the management of the emergency.

[Diagnosis and therapy of gastric carcinoma in the early stage]. / Diagnosi e terapia del carcinoma gastrico allo stadio precoce.

The diagnosis of early gastric cancer is often allowed by endoscopy, which favours the surgical therapy with a five years survival rate of 79-99% of treated patients. Four personal cases are presented, whose evolution differed from their particular gastric localization, associated lesions and surgical treatment. A wide review of the literature is given.

[Prevalence of reactions to drugs in 251 patients with urticaria-angioedema syndrome]. / Prevalenza di reazioni a farmaci in 251 pazienti affetti da sindrome orticaria-angioedema.

We studied 251 outpatients affected by chronic and acute urticaria-angioedema syndrome, triggered or exacerbated by drugs, to evaluate the prevalence of adverse reactions to each pharmacological group. Among these patients, 134 (53.4%) presented one or more adverse reactions to a single drug: 100 (74.7%) reacted to NSAIDs, 33 (24.7%) to antibiotics and 1 to B vitamin complex. The remaining 117 patients (46.6%) presented adverse reactions to more than one drug. Considering the patients all together, 123 (49%) had adverse reactions to ASA, 116 (46.2%) to pyrazones, 65 (25.9%) to antibiotics, 26 (10.3%) to NSAIDs different from ASA and pyrazones. In our experience, according to other reports, there is a greater frequency of drug reactions to NSAIDs vs other drugs. ASA is the more frequently involved drug.

[Diagnostic value and therapeutic limits of endoscopic polypectomy of the stomach]. / Valore diagnostico e limiti terapeutici della polipectomia endoscopica dello stomaco.

Digestive endoscopy represents the current therapy of the gastric polyps. These lesions are endoscopically found in an unexpected number of patients, because of the frequent failure of radiology by itself. Furthermore endoscopic radical polypectomy is mostly possible for the inflammatory and adenomatous polyps. Otherwise endoscopy favours the early detection of malignant changes and coexisting per-cancerous gastropathy. A wide review of the literature is given.

[Surgery in aneurysms of the abdominal aorta in an outlying hospital]. / La chirurgia degli aneurismi dell'aorta addominale in un ospedale periferico.

Abdominal aortic aneurysms have become more frequently recognized due to an improved diagnostic approach and prolonged longevity. Therefore it can be argued that aneurysmal pathology must be entirely dealt with, including its surgical therapy, in outlying non-specialized hospitals. Elective surgical treatment of aortic aneurysms is mandatory in order to acquire the technique that will provide a satisfactory efficiency and reliability in the emergency treatment of ruptured aortic aneurysms. The basic supports of aortic aneurysm surgery in outlying hospitals are the availability of postoperative intensive therapy, a competent surgeon staff and an improved diagnostic approach in order to recognize those aneurysms which are operable by the staff and those requiring very complex reconstructive procedures such as thoraco-abdominal aortic aneurysms which must be transferred to highly specialized centres. An angiography service should be set up since it represents an important support for the operation of abdominal aneurysms; however in practice it proved to be non-essential in the present case. In contrast the intensive postoperative unit must be considered vital in view of the patient's risky conditions. The experience reported here is really positive as regards expanding aneurysms given elective surgery, in which cases there was no mortality either intra- or post-operatively. The follow-up varied from 3 to 7 years. Only one ruptured aneurysm was dealt with and the outcome was fatal owing to anuria caused by severe haemorrhagic shock. However, we cannot expect an acceptable efficiency level in emergency aneurysmectomy if the diagnostic and therapeutic approach does not guarantee full reliability in cases of elective aneurysmectomy.