Evaluation of quality of life in endometriosis patients before and after surgical treatment using the EHP30 questionnaire.

BACKROUND: Endometriosis is one of the most common gynecological illnesses causing extensive psychological, physical and social impact on patient's life and exerts negative effects on health-related quality of Life (HRQoL). However, the effects of surgery on the postoperative HRQoL in the different endometriosis subgroups have not been fully evaluated. METHODS: We performed a comparative retrospective study between 2014 and 2018 at the Medical University of Vienna, including all patients with surgically confirmed endometriosis who had completed the standardized Endometriosis Health Profile-30 (EHP-30) questionnaire 1 day after surgery (the questions refer to the 4 weeks preoperatively) and 6-10 weeks postoperatively. RESULTS: Compared to preoperative values, we found significant benefits, regarding postoperative conditions, in our study group (n = 115) in all five categories, "pain" (HR 0.78, p < 0.001); "self-determination" (HR 0.92, p < 0.001); "emotional health" (HR 0.83, p < 0.001);" social environment" (HR 0.67, p < 0.001); and "self-image" (HR 0.47, p < 0.001). Patients with only peritoneal endometriosis had the lowest preoperative clinical symptoms and there were no significant changes in any of the categories. In the subgroups deep infiltrating endometriosis (DIE) and DIE + ovarian endometrioma, surgical intervention results in a significantly greater improvement in all categories of EHP 30 compared to ovarian endometrioma without DIE or peritoneal endometriosis. CONCLUSION: Our study shows, that especially women with DIE-with or without ovarian endometrioma-demonstrate a more pronounced benefit from surgical therapy compared to patients with peritoneal endometriosis or endometrioma without DIE.

Ultra-sensitive solid-phase Microextraction-Gas Chromatography-Mass spectrometry determination of polycyclic aromatic hydrocarbons in snow samples using a deep cavity BenzoQxCavitand.

A very sensitive and selective solid-phase microextraction-gas chromatography-mass spectrometry method based on the use of a deep cavity BenzoQxCavitand as innovative coating was developed and validated for the simultaneous determination of the 16 US-EPA priority pollutants polycyclic aromatic hydrocarbons (PAHs) in snow samples at ultra-trace levels. The presence of a 8.3 Å deep hydrophobic cavity allowed the engulfment of all the 16 PAHs, providing enhanced selectivity also in presence of interfering aromatic pollutants at high concentration levels. Validation proved the reliability of the method for the determination of the investigated compounds achieving detection limits in the 0.03-0.30 ng/L range, good precision, with relative standard deviations <18% and recovery rates in the 90.8(±2.1)%-109.6(±1.0)%. The detection of low-molecular weight PAHs in snow samples from Antarctica and Alps confirms the widespread occurrence of these compounds, thus assessing the impact of anthropogenic activities onto the environment.

Changes in salivary flow rate and pH in pregnancy.

OBJECTIVE: To evaluate changes in pH and Flow Rate (FR) of the Unstimulated Whole Saliva (UWS) in a sample of pregnant women in different gestational periods. PATIENTS AND METHODS: After collecting demographic data and medical histories, as well as conducting an oral examination, a sample of pregnant women were instructed on how to prepare prior to the sample collection. At a time between 11.00 and 12.00 a.m., they were subjected to salivary collection (spitting method, time 5 minutes); the measurement of FR was carried out using a professionally calibrated precision scale and the pH with a portable pH meter. RESULTS: The average FR of the women's detected sample (0.40 ± 0.20 ml/min) was lower than that of non-pregnant women (0.48 ± 0.15 ml/min) of the same age (p <0.05). We observed an increase (p <0.001) of FR in the first trimester (0.56 ± 0.20 ml/min) compared to second (0.34 ± 0.14 ml/min) and third (0.31 ± 0.14 ml/min) trimester. The salivary pH of pregnant women was lower than the one detected in the non-pregnant women's sample (p <0.0001). CONCLUSIONS: Our study highlighted an increase in the FR in the first trimester compared to that detected in the second and third trimesters of pregnancy which viceversa was lower than the average FR in non-pregnant women. This data, combined with the decrease in salivary pH, supports the hypothesis that correlates the FR increase with the attempt to counter the decrease in pH due to gastric regurgitation frequent in the first trimester. Further studies are necessary to evaluate salivary FR and pH in pregnant women samples related to the emesis phenomenon.

Gestational Diabetes Mellitus: Clinical Characteristics and Perinatal Outcomes in a Multiethnic Population of North Italy.

AIM: To evaluate clinical characteristics and perinatal outcomes in a heterogeneous population of Caucasians born in Italy and High Migration Pressure Countries (HMPC) women with GDM living in Piedmont, North Italy. METHODS: We retrospectively analyzed data from 586 women referring to our unit (2015-2020). Epidemiological (age and country of origin) and clinical-metabolic features (height, weight, family history of DM, parity, previous history of GDM, OGTT results, and GDM treatment) were collected. The database of certificates of care at delivery was consulted in relation to neonatal/maternal complications (rates of caesarean sections, APGAR score, fetal malformations, and neonatal anthropometry). RESULTS: 43.2% of women came from HMPC; they were younger (p < 0.0001) and required insulin treatment more frequently than Caucasian women born in Italy (χ 2 = 17.8, p=0.007). Higher fasting and 120-minute OGTT levels and gestational BMI increased the risk of insulin treatment (OGTT T0: OR = 1.04, CI 95% 1.016-1.060, p=0.005; OGTT T120: OR = 1.01, CI 95% 1.002-1.020, p=0.02; BMI: OR = 1.089, CI 95% 1.051-1.129, p < 0.0001). Moreover, two or more diagnostic OGTT glucose levels doubled the risk of insulin therapy (OR = 2.03, IC 95% 1.145-3.612, p=0.016). We did not find any association between ethnicities and neonatal/maternal complications. CONCLUSIONS: In our multiethnic GDM population, the need for intensive care and insulin treatment is high in HPMC women although the frequency of adverse peripartum and newborn outcomes does not vary among ethnic groups. The need for insulin therapy should be related to different genetic backgrounds, dietary habits, and Nutrition Transition phenomena. Thus, nutritional intervention and insulin treatment need to be tailored.

Fluorinated porous organic frameworks for improved CO2 and CH4 capture.

A porous 3D selectively fluorinated framework (F-PAF1), robust yet flexible and with a surface area of 2050 m2 g-1, was synthesised by condensation of an ad hoc prepared fluorinated tetraphenylmethane (TPM) monomer to ensure homogenously distributed C-F dipoles in the swellable architecture. Tetradentate TPM was also the comonomer for the reaction with fluorinated difunctional monomers to obtain frameworks (FMFs) with a controlled amount of regularly spaced reorientable C-F dipoles. The isosteric heat of adsorption of CO2 was increased by 53% by even moderate C-F dipole insertion, with respect to the non-fluorinated frameworks. CO2/N2 selectivity was also increased up to a value of 50 for the difluoro-containing comonomer. Moreover, methane shows optimal interaction energies of 24 kJ mol-1.

Polypharmacy but not Potential Inappropriate Prescription Was Associated with Frailty in Older Adults from a Middle-Income Country Outpatient Clinic.

OBJECTIVES: the aims of the present study were: (1) investigate the prevalence and association of polypharmacy and pre-frailty or frailty in a middle-income country sample of older adults; and (2) evaluate the prevalence of potential inappropriate prescription (PIP) and its association with pre-frailty or frailty. DESIGN: Cross-sectional observational study. SETTING: Outpatient center at a university-based hospital in the state of São Paulo, Brazil. PARTICIPANTS: 629 older adults from both sexes evaluated between June 2014 and July 2016. MEASUREMENTS: Frailty was identified through the FRAIL scale. All medications received were analyzed by research staff. Presence of PIP was evaluated according to the 2015 updated Beers list. Binary logistic regression tested the association between 4 definitions of polypharmacy (≥ 3, 4, 5, and 6 drugs), and presence of PIP, and the dependent variable pre-frailty and frailty. RESULTS: 15.7% of participants were frail. Polypharmacy was present in 219 (34.8%), and PIP was observed in 184 (29.3%) older adults. All definitions of polypharmacy were significantly associated with frailty (OR between 2.05 to 2.34, p < 0.001). Polypharmacy with 4 or 5 or more drugs were associated with pre-frailty (OR 1.53 and 1.47, respectively). PIP was not associated with frailty (OR 1.47, p = 0.149). CONCLUSIONS: Several definitions of polypharmacy were associated with frailty, but only two were associated with pre-frailty. The presence of PIP was not associated with pre-frailty or frailty.

Adherence to alendronic or risedronic acid treatment, combined or not to calcium and vitamin D, and related determinants in Italian patients with osteoporosis.

PURPOSE: Osteoporosis is a chronic disease and an important health and social burden due to its worldwide prevalence. Literature and clinical experience report incomplete adherence to the therapy. This retrospective observational study aimed at assessing the adherence to first-line antiosteoporosis drugs (AODs; reimbursed by the National Health System, according to the Italian Medicine Agency recommendation number 79), alendronate or risedronate, with or without calcium and/or vitamin D supplements, in a real, Italian clinical setting. PATIENTS AND METHODS: Analyses were carried out on data present in the ARNO Observatory, a population-based patient-centric Italian database. From a population of 5,808,832 inhabitants with available data, a cohort of 3.3 million of patients aged ≥40 years was selected. New users of first-line AODs as monotherapy (accrual period, 2007-2009) were followed up over 3 years to assess adherence at 6, 12, and 36 months to AODs and to supplements and related determinants. RESULTS: Approximately 40,000 new users were identified: mostly women, aged on average (standard deviation) 71±10 years. Alendronate was the most prescribed (38.2% of patients), followed by risedronate (34.9%) and alendronate with colecalciferol as a fixed-dose combination (25.8%). Adherence at the 6-month follow-up was 54%, and this constantly and significantly decreased after 1 year to 46%, and after 3 years to 33% (P<0.01). Adherence to the fixed-dose combination was higher than to plain alendronate throughout the follow-up period. Similarly, adherence to supplements constantly decreased with the duration of treatment. Women and patients aged >50 years were more likely to adhere to treatment regimen (P<0.001). The use of drugs for peptic ulcer and gastroesophageal reflux disease and of corticosteroids for systemic use were significantly associated with high adherence at different times. Polytherapy (>5 drugs), cardiovascular, and neurological therapies were significantly associated with low adherence throughout the follow-up period. CONCLUSION: In a huge clinical practice sample, this study highlights suboptimal adherence to first-line AODs and to supplements and important determinants, such as concomitant therapies.

The role of CSA in the response to oxidative DNA damage in human cells.

Cockayne syndrome (CS) is a rare genetic disease characterized by severe growth, mental retardation and pronounced cachexia. CS is most frequently due to mutations in either of two genes, CSB and CSA. Evidence for a role of CSB protein in the repair of oxidative DNA damage has been provided recently. Here, we show that CSA is also involved in the response to oxidative stress. CS-A human primary fibroblasts and keratinocytes showed hypersensitivity to potassium bromate, a specific inducer of oxidative damage. This was associated with inefficient repair of oxidatively induced DNA lesions, namely 8-hydroxyguanine (8-OH-Gua) and (5'S)-8,5'-cyclo 2'-deoxyadenosine. Expression of the wild-type CSA in the CS-A cell line CS3BE significantly decreased the steady-state level of 8-OH-Gua and increased its repair rate following oxidant treatment. CS-A cell extracts showed normal 8-OH-Gua cleavage activity in an in vitro assay, whereas CS-B cell extracts were confirmed to be defective. Our data provide the first in vivo evidence that CSA protein contributes to prevent accumulation of various oxidized DNA bases and underline specific functions of CSB not shared with CSA. These findings support the hypothesis that defective repair of oxidative DNA damage is involved in the clinical features of CS patients.

Different dynamics in nuclear entry of subunits of the repair/transcription factor TFIIH.

We report here the different ways in which four subunits of the basal transcription/repair factor TFIIH (XPB, XPD, p62 and p44) and the damage recognition XPC repair protein can enter the nucleus. We examined their nuclear localization by transiently expressing the gene products tagged with the enhanced green fluorescent protein (EGFP) in transfected 3T3 cells. In agreement with the identification of more than one putative nuclear localization signal (NLS) in their protein sequences, XPB, XPC, p62 and p44 chimeras were rapidly sorted to the nucleus. In contrast, the XPD-EGFP chimeras appeared mainly localized in the cytoplasm, with a minor fraction of transfectants showing the EGFP-based fluorescence also in the nucleus. The ability of the XPD chimeras to enter the nucleus was confirmed by western blotting on fractionated cell extracts and by functional complementation of the repair defect in the UV5 rodent cells, mutated in the XPD homologous gene. By deletion mutagenesis, we were unable to identify any sequence specific for nuclear localization. In particular, deletion of the putative NLS failed to affect subcellular localization and, conversely, the C-terminal part of XPD containing the putative NLS showed no specific nuclear accumulation. These findings suggest that the nuclear entry of XPD depends on its complexation with other proteins in the cytoplasm, possibly other components of the TFIIH complex.

Study of the bidirectional transport of choline by blocking choline carriers from outside or inside brain nerve terminals.

Membrane carriers can operate bidirectionally. We studied, in rat neocortex synaptosomes, the choline carrier by comparing the ability of the transport inhibitor hemicholinium-3, present outside or inside the nerve terminals, to prevent uptake and release of [(3)H]choline. Because hemicholinium-3 is membrane-impermeable, it was previously entrapped into synaptosomes during homogenization of brain tissue. External and internalized hemicholinium-3 produced similar maximal inhibition (80-90%) of [(3)H]choline uptake. Also comparable (approximately 30 nM) are the potency of externally applied hemicholinium-3 and the estimated potency of the entrapped inhibitor. Exposure to ouabain elicited release of both [(3)H]acetylcholine and [(3)H]choline from synaptosomes prelabeled with [(3)H]choline. The ouabain (300 microM)-evoked release of [(3)H]choline only was blocked by externally added (IC(50) approximately 10 nM) or internalized (estimated IC(50) approximately 5 nM) hemicholinium-3. Release of previously taken up [(3)H]choline elicited by 100 microM external choline (homoexchange) was prevented by external (IC(50) approximately 30 microM) or entrapped (estimated IC(50) approximately 20 microM) hemicholinium-3. The results suggest that the choline carriers fit into the alternating-access model proposed for classical transmitter transport. Entrapping nonpermeant ligands into synaptosomes could allow investigation of the inward-facing conformation of native transporters and how cytoplasmic ligands affect the bidirectional transport of neurotransmitters.

Evaluation of tests employed in serological diagnosis of brucellosis caused by Brucella ovis.

A survey was carried out to verify the sensitivity and specificity of various tests (complement fixation test--CF; agar gel immunodiffusion--AGID; indirect enzyme linked immunosorbent assay--ELISA; immunoblotting--IB) employed in the serological diagnosis of brucellosis caused by Brucella ovis. The tests were executed on 44 blood serum samples of rams coming from B. ovis-free flocks, 75 of B. ovis experimentally infected rams and 1139 from rams living in flocks where B. ovis had been previously isolated. All tests were performed using B. ovis hot saline extract (HS) as antigen. Sensitivity results were 97.4% for IB, 98.68% for CF, 100% for AGID and ELISA; specificity was 100% for all methods. Concordance values were 89.62% (CF-AGID), 78.77% (CF-ELISA), 77.74% (AGID-ELISA), 65.45% (IB-CF), 62.93% (IB-ELISA), 67.24% (IB-AGID). IB identified antibodies to antigenic components with molecular weight of 67, 63, 58, 55, 38, 35, 32, 30, 28, 25, 23, 21, 20-18 kDa (proteins) and 15-12 kDa (rough lipopolysaccharide).

Epididymitis by Brucella ovis: experimental infection in virgin ram lambs.

Ten sexually immature rams were experimentally infected with Brucella ovis, to verify the antibody kinetics and its localization in urinary and genital tracts. All animals became positive to the complement fixation test from the 2nd post infection (p.i.) week and reached the maximum titre (1:256) on the 4th p.i. week. Bacteriemia was demonstrated on 3rd, 4th and 5th p.i. weeks. Two animals, respectively slaughtered 11 and 13 weeks after the infection, showed macroscopic and microscopic genital lesions and the etiologic agent was cultured from their urine and genital organs.

Mutations in the XPD helicase gene result in XP and TTD phenotypes, preventing interaction between XPD and the p44 subunit of TFIIH.

In most cases, xeroderma pigmentosum group D (XP-D) and trichothiodystrophy (TTD) patients carry mutations in the carboxy-terminal domain of the evolutionarily conserved helicase XPD, which is one of the subunits of the transcription/repair factor TFIIH (refs 1,2). In this study, we demonstrate that XPD interacts specifically with p44, another subunit of TFIIH, and that this interaction results in the stimulation of 5'-->3' helicase activity. Mutations in the XPD C-terminal domain, as found in most patients, prevent the interaction with p44, thus explaining the decrease in XPD helicase activity and the nucleotide excision repair (NER) defect.

Leptospira interrogans in the genital tract of sheep. Research on ewes and rams experimentally infected with serovar hardjo (hardjobovis).

To verify if Leptospira hardjo can colonize the male and female genital organs of sheep, 9 animals (6 non pregnant ewes and 3 mature rams) were infected with a strain of L. hardjobovis recently recovered from the kidneys of a seropositive ewe. Postinfection controls (bacteriologic, serologic, immunohistochemistry and electron microscopy) failed to disclose the presence of leptospires in the uterus and oviducts, testicles, epididymis, prostate and bulbourethral glands of animals used for the experiment and slaughtered from 37 to 242 postinfection days. All animals showed a renal localization of L. hardjobovis lasting for the entire period of the study (over 8 months). These results emphasize the important role of sheep as maintenance hosts of the serovar.

Leptospira interrogans serovar hardjo in the kidneys and genital tracts of naturally infected sheep.

A bacteriological study was carried out to identify possible renal and/or genital carriers of Leptospira interrogans serovar hardjo. L. hardjo was found at slaughter in the kidneys of three seropositive ewes, but not in uterus or salpinges of these animals.

Human DNA topoisomerase I-mediated cleavages stimulated by ultraviolet light-induced DNA damage.

DNA topoisomerases have been proposed as the proteins involved in the formation of the DNA-protein cross-links detected after ultraviolet light (UV) irradiation of cellular DNA. This possibility has been investigated by studying the effects of UV-induced DNA damage on human DNA topoisomerase I action. UV lesions impaired the enzyme's ability to relax negatively supercoiled DNA. Decreased relaxation activity correlated with the stimulation of cleavable complexes. Accumulation of cleavable complexes resulted from blockage of the rejoining step of the cleavage-religation reaction. Mapping of cleavage sites on the pAT153 genome indicated UV-induced cleavage at discrete positions corresponding to sites stimulated also by the topoisomerase I inhibitor camptothecin, except for one. Subsequent analysis at nucleotide level within the sequence encompassing the UV-specific cleavage site revealed the precise positions of sites stimulated by camptothecin with respect to those specific for UV irradiation. Interestingly, one of the UV-stimulated cleavage sites was formed within a sequence that did not contain dimerized pyrimidines, suggesting transmission of the distortion, caused by photodamage to DNA, into the neighboring sequences. These results support the proposal that DNA structural alterations induced by UV lesions can be sufficient stimulus to induce cross-linking of topoisomerase I to cellular DNA.

Experimental infection of dogs with Borrelia burgdorferi.

Four beagle dogs were inoculated subcutaneously with the BITs1 Italian strain of Borrelia burgdorferi. Only one dog became infected and B burgdorferi was isolated from its blood and urine three and four weeks after infection. B burgdorferi antibodies were detected by immunofluorescence from four to 11 weeks after infection. An uninoculated dog kept in the same run as the infected dog, developed a positive serological response, but none of the five dogs showed clinical signs.

The ultimate carcinogen of 4-nitroquinoline 1-oxide does not react with Z-DNA and hyperreacts with B-Z junctions.

DNA secondary and tertiary structures are known to affect the reaction between the double helix and several damaging agents. We have previously shown that the tertiary structure of DNA influences the reactivity of 4-acetoxyaminoquinoline 1-oxide (Ac-4-HAQO), the ultimate carcinogen of 4-nitroquinoline 1-oxide (4-NQO), being more reactive with naturally supercoiled DNA than with relaxed DNA. The relative proportion of the three main stable adducts and of an unstable adduct, that resulted in strand scission and/or AP sites, was also affected by the degree of supercoiling of plasmid DNA. In this study we examined the influence of Z-DNA structure on the reactivity of Ac-4-HAQO by mapping the distribution of the two main Ac-4-HAQO adducts, C8-guanine and N2-guanine, along a (dC-dG)16 sequence inserted at the BamHI site of pBR322 plasmid DNA. This insert adopted the left-handed Z and right-handed B structure depending on the superhelical density of the plasmid. Sites of C8-guanine adduct formation were determined by hot piperidine cleavage of Ac-4-HAQO modified DNA, while N2-guanine adducts were mapped by the arrest of the 3'-5' exonuclease activity of T4 DNA polymerase. The results showed that Ac-4-HAQO did not react with guanine residues when the (dC-dG)16 sequence was in Z conformation, while hyperreactivity at the B-Z junction was observed. These results indicate that Ac-4-HAQO can probe the polymorphism of DNA at the nucleotide level.