Psychobiotics Regulate the Anxiety Symptoms in Carriers of Allele A of IL-1ß Gene: A Randomized, Placebo-Controlled Clinical Trial.

BACKGROUND: Probiotic oral intake, via modulation of the microbiota-gut-brain axis, can impact brain activity, mood, and behavior; therefore, it may be beneficial against psychological distress and anxiety disorders. Inflammatory cytokines can influence the onset and progression of several neurodegenerative mood disorders, and the IL-1ß rs16944 SNP is related to high cytokine levels and potentially affects mood disorders. The aim of this study was to examine the combined effect of IL-1ß polymorphism and probiotic administration in mood disorder phenotypes in the Italian population. METHODS: 150 subjects were randomized into two different groups, probiotic oral suspension group (POSG) and placebo control group (PCG), and received the relative treatment for 12 weeks. Psychological profile assessment by Hamilton Anxiety Rating Scale (HAM-A), Body Uneasiness Test (BUT), and Symptom Checklist 90-Revised (SCL90R) was administered to all volunteers. Genotyping was performed on DNA extracted from salivary samples. RESULTS: After 12 weeks of intervention, a significant reduction of HAM-A total score was detected in the POSG (p < 0.01), compared to the PCG. Furthermore, IL-1ß carriers have moderate risk to develop anxiety (OR = 5.90), and in POSG IL-1ß carriers, we observed a reduction of HAM-A score (p = 0.02). CONCLUSIONS: Consumption of probiotics mitigates anxiety symptoms, especially in healthy adults with the minor A allele of rs16944 as a risk factor. Our results encourage the use of probiotics in anxiety disorders and suggest genetic association studies for psychobiotic-personalized therapy.

Developing and cross-validation of new equations to estimate fat mass in Italian population.

OBJECTIVE: Obesity is a global burden that involves more than 500 million people. The objective of this work is to develop and cross-validate the new sex-specific equations to estimate fat mass, based on anthropometric parameters and to compare with other equations. PATIENTS AND METHODS: We evaluated 38762 subjects by dual-energy X-ray absorptiometry (DXA) and enrolled 1434 women and 640 men, aged between 18 and 65 years. Then, we randomized 480 men and 1080 women in developing set and 160 men and 354 women in the cross-validation set. Statistical analysis as multiple regression and Bland-Altman methods were performed. RESULTS: Sex-specific equations were created based on developing set. Then, based on the cross-validating set, these equations were validated and were observed to agree with fat mass by DXA, better than other equations, such as BAI and RFM. CONCLUSIONS: These new sex-specific equations represent an easy tool, since they require only two circumferences, to be used in clinical practice. In the next future, these equations could be validated and refine on specific Italian sub-populations, divided by gender and age, such as the military.

Efficacy and safety of very-low-calorie ketogenic diet: a double blind randomized crossover study.

OBJECTIVE: To verify safety respect to weight loss, cardiometabolic diseases of short-term Very low-calorie ketogenic diets (VLCKDs, <800 kcal day-1). PATIENTS AND METHODS: Randomized cross-over trial with placebo. The study had no. 2 dietary treatment (DT), conducted in two arms: (1) VLCKD1 in which 50% of protein intake is replaced with synthetic amino acids; (2) VLCKD2 with placebo. The VLCKDs (<800 kcal day-1) were different in term of protein content and quality each arm lasted three weeks (wks). Between the two arms a 3-wks washout period was performed to avoid additive effects on DT to follow. At the baseline, at start and end of each arm, all the subjects were evaluated for their health and nutritional status, by anthropometric analysis, body composition (Dual X-ray Absorptiometry (DXA), Bioimpedentiometry, biochemical evaluation, and Peroxisome Proliferator-Activated Receptor γ (PPAR) γ expression by transcriptomic analysis. RESULTS: After VLCKD1 were reduced: Body Mass Index (BMI) (Δ%=-11.1%, p=0.00), Total Body Water (TBW) (p<0.05); Android Fat Percentage (AFP) (Δ%=-1.8%, p=0.02); Android Fat Mass (AFM) (Δ%=-12.7%, p=0.00); Gynoid Fat Mass (GFM) (Δ%=-6.3%, p=0.01); Intermuscular Adipose Tissue (IMAT) (Δ%= -11.1%, p=0.00); Homeostasis Model Assessment of Insulin Re-sistance (HOMA-IR) (Δ%=-62.1%, p=0.01). After VLCKD1 a significant increase of uricemia, cre-atinine and aspartate aminotransferase (AST) (respectively Δ%=35%, p=0.01; Δ%=5.9%, p=0.02; Δ%=25.5%, p=0.03). After VLCKD2 were reduced: BMI (Δ%=-11.2%, p=0.00); AFM (Δ%=-14.3%, p=0.00); GFM (Δ%=-6.3%, p=0.00); Appendicular Skeletal Muscle Mass Index (ASMMI) (Δ%=-17.5%, p=0.00); HOMA-IR (Δ%=-59,4%, p=0.02). After VLCKD2, uricemia (Δ%=63.1%, p=0.03), and Vitamin D levels (Δ%=25.7%, p=0.02) were increased. No significant changes of car-diovascular disease (CVD) indexes were observed after DTs. No significant changes of PPARγ lev-el in any DTs. CONCLUSIONS: 21-days VLCKDs not impair nutritional state; not cause negative changes in global measurements of nutritional state including sarcopenia, bone mineral content, hepatic, renal and lipid profile.

Chronic migraine in women is associated with insulin resistance: a cross-sectional study.

BACKGROUND AND PURPOSE: Migraine is a common neurological disorder. It can be divided into episodic migraine (EM) and chronic migraine (CM), based on headache frequency. Some studies have shown that insulin sensitivity is impaired in migraine; moreover, hypertension, diabetes and obesity are common in patients with CM. The aim of this study was to assess serum glucose, insulin levels and insulin resistance (IR) in a sample of episodic migraineurs, chronic migraineurs and non-pain healthy controls. METHODS: Eighty-three women with EM, 83 with CM and 83 healthy controls were recruited. Headache was diagnosed according to the latest International Classification of Headache Disorders 2 criteria. Waist circumference, body mass index (BMI) and blood pressure were measured. Checked metabolic parameters included fasting glucose, the 2 h 75 g oral glucose tolerance test (2 h OGTT), serum HbA1c, blood lipid profile, C-reactive protein and prolactin. The homeostasis model assessment formula was used to calculate IR. RESULTS: A significant prevalence of IR in CM was observed (P = 0.002). No significant associations were found with fasting glycaemia, the 2 h OGTT, HbA1c, blood lipid profile, C-reactive protein, prolactin and waist circumference. Obesity (BMI >30 kg/m(2)) was associated with an increased risk of CM [odds ratio (OR) 2.4]. When the outcome of interest was the association between IR and obesity, the OR was significantly increased compared with IR alone (OR = 13.2). CONCLUSION: This may suggest that CM is associated with IR status, particularly when it is in partnership with obesity.

Association between -308 G/A TNF-α polymorphism and appendicular skeletal muscle mass index as a marker of sarcopenia in normal weight obese syndrome.

BACKGROUND AND AIM: Normal weight obese (NWO) syndrome is characterized by normal body mass index (BMI), but high amount of fat mass and reduced lean mass. We evaluated allelic frequency of the G/A -308 TNF-α polymorphism and prevalence of sarcopenia in NWO. METHODS: We enrolled 120 Italian healthy women, distinguished into 3 groups: normal weight (NW); NWO, and preobese-obese (PreOB/OB) and evaluated anthropometric parameters, body composition by dual X-ray absorptiometry, blood tests, and genotyping of G/A -308 TNF-α polymorphism. RESULTS: We found a positive association between sarcopenic obesity and -308 TNF-α polymorphism. All obese women were sarcopenic and were no carrier of mutation (G/G). Among all G/G, NWO showed significant differences in lean mass and total body lean mass (TBLean) with respect to NW and PreOB/OB (P < 0.001). Regarding appendicular skeletal muscle mass index values, 4.21% of NW were sarcopenic (50% G/G and 50% G/A); the same percentage was observed in NWO subjects (100% G/G). Moreover, 2.10% of PreOB/OB were sarcopenic and all were G/G. CONCLUSION: Our study suggests that TNF-α polymorphism contributes to sarcopenic obesity susceptibility, in association with body composition. This is the first study that shows the importance of TNF-α polymorphism to determine TBLean variation in NWO syndrome.

Urrets- Zavalia Syndrome after glaucoma filtration device implantation.

We report a case of Urrets-Zavalia Syndrome after a glaucoma filtration device (g.f.d.) implantation. A 74-year-old woman with bilateral advanced glaucoma has been planned for surgery. The patient underwent to g.f.d. implantation in the right eye. On postoperative day 1, the patient had an edematous cornea with a dilated and non reactive pupil. In this article we describe the clinical history of this patient. To our knowledge, this is the first case of Urrets-Zavalia Syndrome after a g.f.d. implantation.

Body composition phenotype: Italian Mediterranean Diet and C677T MTHFR gene polymorphism interaction.

OBJECTIVES: Strategies to improve weight maintenance are focused on considering the genetic makeup and its interaction with dietary intake, with the aim to identify vulnerable individuals that will benefit from a variety of more personalized dietary recommendations. The aim of the study was to examine the impact of the C677T MTHFR gene Polymorphism on body composition changes induced by a balanced hypocaloric Italian Mediterannean diet (IMD). SUBJECTS AND METHODS: Participation in the study included a complete screening of anthropometry and body composition by Dual-energy X-ray absorptiometry (DXA), and a genotyping for the C677T MTHFR polymorphism. 70 Italian Caucasian obese were enrolled and 56 of them completed the screening at baseline and 12 weeks after the nutritional intervention. RESULTS: T(+) carriers had a higher content of Total Body Fat (TBFat), and Lean (TBLean), reflecting on higher weight and BMI, than T(-) carriers. After IMD, the 28.6% and 71.4% of total subjects decreased weight and TBFat (Kg), respectively. The relative changes were: delta % = -9.09±3.85 for weight; delta % = -15.79±8.51 for TBFat; delta % = -3.80±5.60 for TBLean. The 5.3% of subjects who reached the end point of intervention, and the 8.9% who reduced TBFat (%) below the cut-off of preobesity, were T(-) carriers. A loss of TBLean (Kg) was observed in the 5.1% and 23.5% of T(-) and T(+) carriers. CONCLUSIONS: MTHFR genetic variations analysis would be an innovative tool for the nutritional assessment, in order to predict the therapeutic response of obese subjects, in terms of fat and lean mass loss.

Effect of MN (III) tetrakis (4-benzoic acid) porphyrin by photodynamically generated free radicals on SODs keratinocytes.

Superoxide, a reactive form of oxygen, can be produced in vivo either in normal and under pathophysiologic conditions or by photosensitizing chemicals, as during photodynamic treatment. Photodynamic therapies (PDT), widely adopted in Dermatology and Oncology, are known to generate reactive oxygen species (ROS) and may contribute to structural alterations and oxidatively generated modifications of cellular antioxidants. We hypothesized that over-production of free radicals would decrease the enzymatic activities of endogenous cellular antioxidants. To test this hypothesis, keratinocytes were treated with the photosensitizer Photofrin plus visible light to produce free radicals and CuZnSOD and MnSOD activities were measured. Photodynamic treatment of keratinocytes increases malonylaldehyde production, nitrotyrosine staining and superoxide production. The enzymatic activities of CuZnSOD and MnSOD were significantly decreased after Photofrin plus visible light treatment. Our results suggest that the main cellular antioxidant system can be inactivated by photodynamically generated ROS. Pretreatment of keratinocytes with free radicals scavenger such as Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) was able to restore the endogenous antioxidant system activities, inhibiting the MDA formation, nitrotyrosine staining and superoxide formation. Antioxidant therapy could therefore be a useful tool in protecting healthy epidermal cells against common side effects induced by antitumor targeted therapies.

Effects of dark chocolate in a population of normal weight obese women: a pilot study.

BACKGROUND: Normal weight obese (NWO) syndrome is defined as an excessive body fat associated with a normal body mass index and characterized by a higher risk for cardiovascular morbidity and mortality. Recent studies have demonstrated that dark chocolate (DC) has beneficial effects in the prevention of cardiovascular diseases (CVD) due to its anti-inflammatory and antioxidant properties. AIM: The aim of the present study was to investigate the effects of DC consumption on lipid profile, inflammatory markers, biochemical parameters, and blood pressure, in NWO women. MATERIALS AND METHODS: 15 women affected by NWO syndrome, aged 20-40 years, were included in the study. After a DC-free washout period, subjects received DC (100 g/die) containing 70% cocoa for 7-days. Body composition by Dual energy-X-ray absorptiometry (DXA) was performed at baseline. Blood pressure, anthropometric measurements, biochemical parameters and plasma levels of some cytokines were measured before and after DC consumption. RESULTS: After DC consumption, we observed a significant increase in the HDL cholesterol level (Delta% = +10.41±13,53; p ≤ 0.05), a significant decrease of total cholesterol/HDL cholesterol ratio (Delta %= -11.45±7.03; p ≤ 0.05), LDL/HDL cholesterol ratio (Delta % = -11.70±8.91; p ≤ 0.05), and interleukin-1 receptor antagonist (IL-1Ra) (Delta % = -32.99±3.84; p ≤ 0.05). In addition, a reduction in abdomen circumference was observed. We also found a positive correlation between changes in atherogenic indices, and IL-1Ra, abdomen reduction. CONCLUSIONS: Our findings suggest that regular consumption of DC could be useful in maintaining a good atherogenic profile, due to the favourable effects on HDL cholesterol, lipoprotein ratios and inflammation markers.

Effects of paraquat and capsaicin on the expression of genes related to inflammatory, immune responses and cell death in immortalized human HaCat keratinocytes.

The present experiments were aimed to characterize in immortalized human HaCat keratinocytes the gene expression induced by paraquat and capsaicin, two agents known to induce cell death or to affect inflammatory and pain pathways, respectively. In particular, the following set of genes were analysed by qRealtime PCR: CXCL10,CXCL11, IL-10 (inflammatory and immune responses), TP73, BCL2, (apoptotic and anti-apoptotic genes), MMP9 (proteolysis), SOD-1, BAK-1 and CAT (peroxysomal and microsomal oxidation pathways). In this way, we were able to differentiate the two toxins since they had a different profile of gene expression. In fact, paraquata was found to activate set of genes involved in inflammatory (CXL10,CXL11 and IL-10), and cell death (BCL2, BAK-1, MMP9) pathways. Another specific site of action of paraquat was represented by an activation of the gene involved in SOD-1 transcription. On the contrary, capsaicin was found to produce only an up-regulation of BCL2, an anti-apoptotic gene and MMP9, whereas no significant changes were reported in genes involved in inflammatory and immune responses. Finally, in comparison to previous experiments carried out with TNF-alpha and IL-1beta, we have shown that paraquat produced a similar pattern of activation of set of genes involved both in inflammation and apoptosis.

DJ-1 is a Parkinson's disease susceptibility gene in southern Italy.

Mutations in the gene DJ-1 have been shown to be a rare cause of early-onset Parkinson's disease (EOPD). Since DJ-1 mutations have been found in patients with Parkinson's disease (PD) from southern Italy, we aimed to investigate whether polymorphisms within the DJ-1 gene could represent a risk factor for sporadic PD. First, we genotyped 294 patients with PD and 298 controls coming from southern Italy to assess the distribution of the insertion/deletion (Ins/Del) polymorphism. In a second phase, we identified five single-nucleotide polymorphisms (SNPs) useful to delimit a region potentially involved and genotyped all patients and controls for these markers. All the markers analyzed were significantly associated with PD at both allelic and genotypic level. The most significant association with the disease was found at the Ins/Del polymorphism (p = 0.0001; adjusted odds ratio (OR ) = 2.05; confidence interval (CI ) = 1.36-3.08). When we considered a three-marker sliding window, we found a highly significant association between the disease and the haplotypes including markers rs17523802, Ins/Del, and rs3766606 (p = 0.0007) and markers Ins/Del, rs3766606 and rs7517357 (p = 0.0054). Our results indicate that polymorphisms located in a region spanning 3535 bp from the promoter to the intron 2 of the DJ-1 gene confer risk to sporadic PD in southern Italy.

Caspase-1 inhibitors abolish deleterious enhancement of COX-2 expression induced by HIV-1 gp120 in human neuroblastoma cells.

The human CHP100 neuroblastoma cell line has been shown to provide an useful in vitro model to elucidate the mechanisms underlying HIV-1 gp120 neurotoxicity. Here we report western blotting evidence demonstrating that exposure to a cytotoxic concentration of the viral coat protein up-regulates expression of the inducible isoform of cyclooxygenase (COX-2) in neuroblastoma cells and this seems to be due to the previously observed increase in secreted IL-1beta. In fact, here we show that acetyl-Tyr-Val-Ala-Asp-chloromethylketone (Ac-YVAD-CMK) and t-butoxycarbonyl-L-aspartic acid benzyl ester-chloromethylketone (Boc-Asp-(OBzl)-CMK), two inhibitors of Interleukin-1 Converting Enzyme (ICE; also referred to as caspase-1), abolish COX-2 expression enhanced by gp120 and consequent cell death. In addition, NS-398, a selective inhibitor of COX-2 activity, affords neuroprotection strengthening the role of COX-2 in the mechanisms of death. In conclusion, the present data support the notion that IL-1beta is the signal through which gp120 elevates COX-2 expression and the latter is strongly implicated in the mechanisms underlying cytotoxicity.