Distribution of selected trace elements in the major fractions of donkey milk.

The aim of this study was to evaluate the concentrations of Zn, Cu, Mn, Se, Mo, Co, Li, B, Ti, Cr, Rb, Sr, Cd, and Pb in donkey milk and their distribution in major milk fractions (i.e., fat, casein, whey proteins, and aqueous phase). Individual milk samples were provided by 16 clinically healthy lactating donkeys. Subsequent centrifugation, ultracentrifugation, and ultrafiltration were carried out to remove fat, casein, and whey proteins to obtain skim milk, a supernatant whey fraction, and the aqueous phase of milk, respectively. Concentrations of the elements were measured in whole milk and fractions by inductively coupled plasma-mass spectrometry, and the concentrations associated with fat, casein, and whey proteins were then calculated. The effect of removal of fat, casein, and whey proteins was determined by repeated-measures ANOVA. The fat fraction of donkey milk carried a small (∼4.5% to 13.5%) but significant proportion of Mo, Co, Ti, Cr, and Sr. The casein fraction in donkey milk carried almost all milk Zn, a majority of Cu and Mn, and most of Mo, Ti, and Sr. Relevant proportions, between 20% and 36%, of Se, Co, and Cr were also associated with caseins. The majority of Se, Co, Li, B, Cr, and Rb, and relevant proportions of Mn, Mo, Ti, and Sr were found in soluble form (ultracentrifuged samples) and distributed between whey proteins and the aqueous phase of milk (ultrafiltered samples). Whey proteins in donkey milk carried the majority of milk Se and Co. All Li and B was present in the aqueous phase of milk, which also contained most Rb and Cr, and 17% to 42% of Mn, Se, Mo, Co, Ti, and Sr.

Development of a multivariable prediction model to identify dairy calves too young to be transported to auction markets in Canada using simple physical examination and body weight.

Calves born on Eastern Canadian dairy farms that are not kept in the herds are traditionally sold through auction markets and are raised for meat purposes such as veal calves. Since February 2020, a new Canadian federal regulation has forbidden calves <9 d old to be sold through auction markets. However, in the absence of a real-time birth registry consultation system, it would be of interest to look for predictors that could be associated with age to allow identification of calves too young to be transported. In the current retrospective cross-sectional study, 1,178 calves with a declared birth date (411 calves aged <9 d old; 34.9%) were assessed in 2 large Québec auction sites. Easy-to-record covariates [body weight (BW), breed phenotype, and presence of an umbilical cord remnant] as well as other clinical signs (umbilical swelling, enlargement, umbilical pain, wet umbilicus, skin tent, sunken eyes, ocular and nasal secretion, and hide cleanliness) were assessed. Two logistic regression models using age as a dichotomous dependent variable (<9 d old vs ≥9 d old) were built. The first model (model 1) considered all covariates, which were selected after univariable analyses and a backward stepwise selection process, whereas a more pragmatic model (model 2) only included the 3 easy-to-record variables (i.e., BW, breed, umbilical cord). Both models had similar accuracy to detect calves <9 d old (sensitivity of 38.4 and 37.5%, and specificity of 85.7 and 84.6% for model 1 and 2, respectively). Model 2 was subsequently more specifically studied as it employs a faster and easier assessment. Decision thresholds were tested for their robustness based on misclassification cost term (MCT) analysis with various prevalence of calves <9 d old and various costs of false-negative:false-positive ratio. Despite statistical significance, model accuracy, even if refined with MCT analysis, was limited at the individual level, showing the limits of using physical signs and BW or their combination as a reliable proxy of age. The sensitivity of these models to find calves <9 d old was not to be used for monitoring compliance with the Canadian federal regulation. The relatively high model specificity may help to use this model as a rule-in test (i.e., targeting positive calves for further investigation) rather than a rule-out test (due to its low sensitivity).

Distribution of calcium, phosphorus, sulfur, magnesium, potassium, and sodium in major fractions of donkey milk.

The aim of this study was to evaluate the concentrations of Ca, P, S, Mg, K, and Na, and their distribution in major fractions of donkey milk (i.e., fat, casein, whey proteins, and aqueous phase). Individual milk samples were collected by mechanical milking from 16 clinically healthy lactating donkeys. Milk yield per milking was recorded and milk gross composition, casein content, and pH were determined. Whole milk samples were centrifuged to separate fat and to obtain skim milk. Skim milk samples were ultracentrifuged to separate a sedimentable casein pellet and to obtain a supernatant whey (soluble) fraction, which was then ultrafiltered to obtain the aqueous phase of donkey milk. Whole milk and the processed samples were analyzed for the aforementioned elements by inductively coupled plasma-mass spectrometry. The concentration of elements associated with fat, casein, and whey proteins was then calculated. All the Na was present in the aqueous phase. The fat fraction in donkey milk carried very little or none of the investigated elements. The majority of Ca (62.9%) and P (53.1%) was associated with casein, and the rest of these elements was mostly present in the aqueous phase. The majority of Mg was present in the aqueous phase, but a relevant part (32.6%) was associated with the casein fraction. No K was associated with casein. On a molar basis, the ratio of colloidal Ca and P to casein (mmol/g of casein) was more than double the values reported in literature for cow milk. The correlation coefficient was negative between milk pH and P in the ultracentrifuged (r = -0.81) and ultrafiltered (aqueous) fraction (r = -0.66). Milk pH correlated positively with colloidal Ca (r = 0.59) and with the ratio of colloidal Ca to casein (mmol/g of casein; r = 0.68). Colloidal Ca and P were positively correlated (r = 0.64). These data suggest that the high ratio of colloidal Ca and P to donkey casein micelles is due to a larger amount of colloidal calcium phosphate bound to casein micelles compared with literature data on cow milk. The percentage of elements associated with whey proteins was less than 5% for Ca, P, and K, but Mg reached approximately 9% of total Mg. The majority of S (63.6%) was associated with whey proteins, and only one-fourth of this element was associated with casein, indicating a higher content of sulfur-containing amino acids in donkey whey proteins than in casein.

Shoulder periprosthetic fracture in elderly patient: a minimally invasive osteosynthesis and "off-label" treatment with teriparatide. A case report and literature review.

A case of shoulder periprosthetic fracture in elderly patient. The patient underwent a minimally invasive osteosynthesis and "off-label" treatment with teriparatide. An 80-year-old woman patient following an accidental fall reported a transverse displaced diaphyseal fracture of the right humerus, distal to the stem of the inverse prosthesis. The patient suffering from severe osteoporosis and chronic ischaemic heart disease. The patient underwent fracture osteosynthesis surgery using a Hoffmann III mono-axial external fixator. Teriparatide administered at a dosage of 20 micrograms/day, for four months. At six months from the beginning of th e hybrid treatment, a complete healing of the fracture was observed radiologically and clinically. It is possible to affirm that the use of teriparatide off-label has a positive and additive effect in promoting the healing of fractures.

Breast cancer and thyroid diseases: analysis of 867 consecutive cases.

INTRODUCTION: The aim of this study was to evaluate the association of breast cancer with thyroid diseases. Many authors have already studied the possible relationship between these two diseases, but the results are nowadays conflicting. MATERIALS AND METHODS: Our study examining 867 patients with breast cancer (862 women and 5 men, average age = 61 years old) of whom 141 also had thyroid disease evaluated the recurrence of thyroid diseases and their association with different types of breast cancer. Statistical analyses were performed using SPSS software for Windows; we used nonparametric tests (Chi-square and Mann-Whitney), and the level of significance was set at p < 0.05. RESULTS: We found a significant association between breast cancer diagnosed after menopause and thyroid disease (p < 0.003). Moreover, we analyzed the role of thyroid autoimmunity identifying an association between chronic autoimmune thyroiditis and breast cancer diagnosed before menopause (p < 0.05). Regarding receptor profile of breast carcinoma, we have found an increased expression of estrogen receptors in patients with autoimmune thyroiditis compared to patients with any other thyroid diseases (p < 0.03). Contrariwise, we do not have found any difference between the group with every thyroid disease and the group without thyroid disease (p < 1.00). We did not find other statistically significant associations with breast cancer's parameters like family history, tumor size, lymph node metastasis, distant metastasis, cancer clinical and pathological stage, differentiation grade and expression of Ki67, p53 and Her2 in the two main groups with or without thyroid disease. Likewise, we did not found other statistically significant association between hypothyroidism or hyperthyroidism and breast cancer.

Central olfactory processing in patients with disorders of consciousness.

BACKGROUND AND PURPOSE: Previous studies have demonstrated that individuals suffering from disorder of consciousness (DOC) maintain some minor neural processing of percepts mediated by senses that early in their pathway intersect the thalamus, a key dysfunctional area in DOC patients. Here the degree of sensory preservation within the olfactory system, a system that lacks an obligatory thalamic relay, and its relationship to the consciousness level in DOC patients of various etiologies was assessed. METHODS: Clinical Coma Recovery Scale - Revised (CRS-R) as well as cerebral responses to odors by means of functional magnetic resonance were obtained in a group of vegetative state/unresponsive wakefulness syndrome (n = 26) patients, minimally conscious state (n = 7) patients and healthy controls (n = 25). RESULTS: A majority of vegetative state/unresponsive wakefulness syndrome patients (58%) and 100% of minimally conscious state patients demonstrated a significant preservation of olfactory neural processing, manifested by activation within the piriform cortex, an area considered as a primary olfactory region. Degree of preservation of olfactory processing differed linearly in line with the patients' etiologies where groups demonstrating greater conscious awareness demonstrated more significant processing. Viewed over all DOC patients, there was a significant negative association between odor-related activity in the orbitofrontal cortex and CRS-R scores. CONCLUSIONS: It is demonstrated that DOC patients exhibit a significant preservation of olfactory neural processing with a clear relationship to etiopathologies and clinical measures even years after of chronification of DOC.

Resting state fMRI in cluster headache: which role?

The pathophysiology of cluster headache (CH) is not well-known. For several years, the most widely accepted theory was that CH was triggered by hypothalamus with secondary activation of the trigeminal-autonomic reflex. However, it was recently suggested that the posterior hypothalamus might be an actor of the pain modulating network more involved in terminating rather than triggering attacks. To investigate this hypothesis, resting state fMRI could provide valuable information on functional connectivity between brainstem and hypothalamus, as well as other brain structures that could be involved in CH pathophysiology. In this framework, here we review recent studies investigating functional connectivity by means of resting state fMRI. Despite the important findings of these studies, we suggest that important steps in the comprehension of CH pathophysiology will be done when the scientific community will use the new methodological approaches recently suggested to study functional connectivity in the brainstem.

Minor and potentially toxic trace elements in milk and blood serum of dairy donkeys.

The aim of this trial was to study the concentration of Ti, V, As, Rb, Sr, Mo, Cd, Cs, and Pb in donkey milk and blood serum. One hundred twelve individual milk and blood serum samples were collected from 16 lactating donkeys (Martina-Franca-derived population; 6 to 12 yr old; 3 to 7 parities; average live weight 205.4kg; 32 to 58 d after foaling at the beginning of the trial) during a 3-mo-long experiment. The samples were analyzed for the aforementioned elements by inductively coupled plasma-mass spectrometry. Feedstuff and drinking water were also analyzed for the investigated elements. Data were processed by ANOVA for repeated measures. Average milk concentrations (±SD) of Ti, Rb, Sr, Mo, Cs, and Pb were 77.3 (±7.7), 339.1 (±82.1), 881.7 (±270.4), 4.5 (±1.6), 0.49 (±0.09), and 3.2 (±2.7) µg/L, respectively. More than 80% of samples were below the limit of detection for V, As, and Cd in milk and for Cd, and Pb in blood serum. The lower bound calculated for milk V, As, and Cd was 0.03µg/L for the 3 elements, the upper bound was calculated at 0.23, 0.10, and 0.31µg/L and the maximum value was observed at 0.54, 0.15, and 0.51µg/L, respectively. The average milk concentrations of Ti, Rb, Sr, Mo, and Cs were 600, 458, 346, 16, and 294%, respectively, than those of blood serum. Yet, Cs concentrations were in the same order of magnitude in milk and serum. Moderate to strong positive and significant correlation coefficients were observed between milk and blood serum concentrations for Ti, Rb, Sr, and Cs. The effect of the stage of lactation was significant for all the investigated elements in milk and blood serum, but most of the elements showed only small changes or inconsistent trends, and only the concentrations of Rb and Sr showed decreasing trends both in milk and blood serum. The relationship between milk and blood serum element concentrations indicates that the mammary gland plays a role in determining the milk concentrations of Mo, Ti, Rb, Sr, Mo, and Cs. In the current experimental conditions, in agreement with the low levels in drinking water and feedstuff, donkey milk concentration of potentially toxic elements was very low and did not raise health concerns for human consumption.

Pulmonary inflammation and cell death in mice after acute exposure to air particulate matter from an industrial region of Buenos Aires.

Epidemiological studies have shown that air particulate matter (PM) can increase respiratory morbidity and mortality being the lungs the main target organ to PM body entrance. Even more, several in vivo and in vitro studies have shown that air PM has a wide toxicity spectra depending among other parameters, on its size, morphology, and chemical composition. The Reconquista River is the second most polluted river from Buenos Aires, and people living around its basin are constantly exposed to its contaminated water, soil and air. However, the air PM from the Reconquista River (RR-PMa) has not been characterized, and its biological impact on lung has yet not been assessed. Therefore, the present investigation was undertaken to study (1) RR-PMa morphochemical characteristic and (2) RR-PMa lung acute effects after intranasal instillation exposure through the analysis of three end points: oxidative stress, inflammation, and apoptosis. A single acute exposure of RR-PMa (1 mg/kg body weight) after 24 h caused significant (p < 0.05) enrichment in bronchoalveolar total cell number and polymorphonuclear (PNM) fraction, superoxide anion generation, production of pro-inflammatory cytokines TNF-α and IL-6, and induction of apoptosis. It was also observed that in lung homogenates, none of the antioxidant enzymes assayed showed differences between exposed RR-PMa and control mice. These data demonstrate that air PM from the Reconquista River induce lung oxidative stress, inflammation, and cell death therefore represents a potential hazard to human health.

Delayed graft dislocation after thoracic aortic endovascular repair.

BACKGROUND: Endograft dislocation in thoracic aorta has not been widely studied. OBJECTIVES: The purpose of this study is to analyze the incidence and predisposing factor in a single centre experience after 117 procedures. METHOD: Between November 2000 and December 2011, all consecutive patients undergoing endovascular repair for descending thoracic or thoraco-abdominal aortic disease were identified. Follow-up imaging protocol included triple-phase CT-angiography at 1, 4, and 12 months, and annually thereafter. Migration was defined as proximal/ distal movements >10 mm relative to anatomical landmarks or any movement leading to symptoms or reintervention. RESULT: We identified 117 patients. Mean follow-up was 32 months (range, 1-144). Overall, five (4.3%) patients with thoracic EG dislocation were identified. Dislocation was classified as collapse/infolding in 3 cases and migration in 2. Mean delay of the dislocation was 12.7 months. Only one patient developed symptoms and required an additional endograft. In the group of dislocated endografts, mean age (53 ± 20 vs. 68 ± 15, P = .032) and the diameter of the aortic lesion were lower (4.1 cm ± 1.6 vs. 5.6 cm ± 1.8, P = .069), and the proximal landing zone at "zones 2 and 3" were more frequently used (5 vs. 65, P = 0.81). All but one patient with collapse/infolding are still alive and doing well at a mean follow-up of 80 months. CONCLUSION: Dislocation is an infrequent complication, but not so rare. Young age, small aortic diameter , and proximal sealing at the distal arch were the most important data associated with this complication.

Could serum procalcitonin play a role in an emergency setting for patients with pyogenic spondylodiscitis?

OBJECTIVE: Spinal infections, represent quite rare but often severe conditions. However, due to symptoms' non-specificity and the lack of specific laboratory tests, diagnosis is often delayed with serious consequences for the patient's outcomes. The present investigation aimed at evaluating the role of procalcitonin (PCT) and other clinical features on the risk stratification and the clinical outcomes in spondylodiscitis patients treated in our Emergency Department. PATIENTS AND METHODS: The present investigation represents a single-center retrospective study. Clinical records of consecutive patients admitted to our Emergency Department from 1 January 2015 to 31 March 2021 were evaluated and patients with spondylodiscitis diagnosis in this period were recruited. Our primary outcome was the degree of autonomy of patients following the acute event. Our secondary outcome was the resolution of the infection. RESULTS: In the study period, a total of 345 patients were evaluated. Among these, 165 met the inclusion criteria, and constituted the study cohort. Concerning the primary outcome, we observed that the most significant predictive factors for being non-autonomous were elevated serum creatinine (> 1.05 mg/dl), Blood Urea Nitrogen (BUN) > 23 mg/dl, Lactate dehydrogenase > 228 U/L, PCT > 0.11 ng/mL. Patients with higher PCT (PCT > 0.11 ng/mL) and higher BUN (BUN > 23 mg/dl) had higher odds of infection persistence (the Odd Ratio, OR, were respectively 3.78 for PCT and 3.14 for BUN). CONCLUSIONS: PCT assay may play a role in diagnosing spondylodiscitis in an emergency setting. A PCT value > 0.11 ng/mL should be considered as a red flag, a predictor of worse clinical outcomes and persistence of infection.

An extraperitoneal approach for complex flank, iliac, and lumbar hernia.

PURPOSE: The aim of this paper is to propose our four-step technique, an open extraperitoneal approach for complex flank, lumbar, and iliac hernias. METHODS: A big polypropylene mesh is placed, covering and reinforcing all the lateral abdominal wall in an extraperitoneal space. Its borders are retroxiphoid fatty triangle and the costal arch cranially and the retropubic space caudally, psoas muscle, and paravertebral region posteriorly and contralateral rectus muscle medially. Mesh dimensions do not depend from the defect size, but prosthesis has to cover all the lateral abdominal wall. RESULTS: No major complications have been reported. The mean length of stay is 4.8 days (range 3-11). Mean follow-up is 44.8 months (range 5-92). One recurrence (4.5%) has been reported at the 1-year clinical evaluation. CONCLUSION: In conclusion, we believe that regardless size and location of the defect, every complex lateral hernia requires the same extensive repair because of the critical anatomy of the region with a big medium-heavyweight polypropylene mesh placed in an extraperitoneal plane, the only one that allows adequate covering of the visceral sac. Our technique is a safe, feasible, and reproducible treatment for this challenging surgical problem.

Sensorineural hearing loss in ankylosing spondylitis treated with TNF blockers.

OBJECTIVES: Sensorineural hearing loss (SNHL) in the uncommon disease ankylosing spondylitis (AS) has been previously reported. We analysed the relationships between AS, anti-TNF drug treatment, and SNHL. METHODOLOGY: We determined pure tone thresholds in 28 consecutive patients with AS who were treated with the TNFalpha inhibitors etanercept or infliximab, or with a TNFalpha inhibitor plus methotrexate (MTX). RESULTS: SNHL was diagnosed in 16 patients (57.1%): 7/7 (100%) of those treated with anti-TNFalpha plus MTX, and 9/21 (43%) of those treated with anti-TNFalpha alone. We found a significant association between SNHL and treatment modality (p = 0.011) or treatment time in months (p = 0.020). CONCLUSIONS: The SNHL rate was significantly higher in patients treated with anti-TNFalpha plus MTX than those treated with anti-TNFalpha alone. The culpability of anti-TNF therapy was supported by the association between SNHL and treatment time, which was longer for anti-TNFalpha plus MTX than for anti-TNFalpha alone. SNHL may be due not only to AS, but also to drug-induced vasculitis of the labyrinthine artery or its cochlear branch.

Functional-MRI evaluation of pain processing in chronic migraine with medication overuse.

Withdrawal is the first step for treating patients with chronic migraine and medication overuse. Recent studies confirmed common elements in personality between these patients and subjects addicted; some neuroimaging researches showed that abnormalities revealed are related to a specific cerebral pattern and that they can return to the normal state after withdrawal. Aim of the study was to submit a group of patients suffering from chronic migraine and medication overuse (the diagnosis was made according to Silberstein-Lipton criteria) to a withdrawal, to evaluate by f-MRI the presence of specific cerebral patterns before treatment and their possible changes after withdrawal. f-MRI seems to be a useful technique to obtain information on particular neuronal changes of the pain network involved in this type of patients. The activated areas are congruent with some data of the literature and the data emerged are discussed according to preceding reports.

[A particular anthropometric method for the study of accessibility of a workstation]. / Un particolare approccio antropometrico per lo studio della raggiungibilità di una postazione cassa.

One of the main factors which can involve musculo-skeletal disorders is the assumption of awkward postures. These lasts can be caused, in some cases, by a no-suitable collocation of some devices which are indispensable for the work. It is possible to evaluate if the chosen collocation is adequate or not by studying the accessibility of the workstation with a special regard for the accessibility of the devices placed inside the workstation. EN ISO 14738:2002 is a specific standard which has been adopted in Italy as UNI EN ISO 14738:2004. This standard gives some useful requirements, in terms of accessibility, to design a workstation at no-mobile machinery. In this study, the authors have analyzed a check out workstation by following the requirements described in UNI EN ISO 14738:2004. Critical aspects, related to the organization both of the work activities either of the workstation, have been highlighted taking into account standard criteria. Finally the authors make a new design of the check out workstation trying to optimize device collocation in order to reduce awkward postures. The new configuration has been investigated by applying the criteria mentioned in the standard.

Assessment of systematic reviews and meta-analyses available for bovine and equine veterinarians and quality of abstract reporting: A scoping review.

OBJECTIVE: Evidence-based medicine (EBM) is used in various areas including veterinary medicine. The assessment of the quality of systematic review and meta-analyses (SR-MA) despite their importance in the EBM process is uncommonly performed in veterinary medicine due to the absence of specific dedicated tools. The main objective of this observational study was to examine the extent and nature of SR-MA that can be available online to an equine or bovine veterinarian. Secondary objectives included: (1) to determine if A MeaSurement Tool to Assess systematic Reviews (AMSTAR) tool can be used for equine and bovine SR-MA methodological assessment and if it has a satisfactory interrater reliability in a subsample of these SR-MA and (2) to appraise the completeness of abstract reporting of this sample. STUDY DESIGN AND SETTING: A scoping review using equine and bovine medical science SR-MA retrieved from PubMed was performed. A sub-sample of these reviews (n = 30) were independently assessed by 3 different raters using the AMSTAR tool validated for medical reviews. The completeness of abstract reporting was performed using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) checklist. RESULTS: Ten and 168 SR-MA were retrieved for equine and bovine species respectively. For bovine SR-MA, 93 reviews were about nutrition and metabolism topic. On the 30 SR-MA subsamples, with 10 equine and 20 bovine SR-MA randomly chosen for AMSTAR assessment, the median interrater agreement (Kappa) was 0.60 (interquartile range: 0.36-0.71) depending on AMSTAR item and pairs of raters. When focusing on the total score of AMSTAR, the inter-observer intra-class correlation coefficient was very good (0.85, 95% confidence interval (CI): 0.74-0.92). The AMSTAR items that were unfrequently reported (33% or less of reviews) were "a priori" protocol of SR-MA specification, complete report of the list of studies (included and excluded studies), quality assessment of the included studies, publication bias assessment and conflict of interest (reported either for included studies and for SR-MA authors). Abstracts reporting quality was low with a median percentage of complete reported items of 33% (range: 8-58%) CONCLUSIONS: In large animal veterinary medicine, SR-MA are uncommonly performed in equine and bovine medicine. The SR-MA can be assessed using AMSTAR with acceptable inter-rater reliability, which is helpful to assess SR-MA methodological quality.

[Function analysis of employees at cash desks in the retailing sector by means of the study of the movements of the upper limbs]. / Analisi della mansione dell'addetto alle casse nella grande distribuzione mediante lo studio del movimento degli arti superiori.

Supermarket checkouts are different in term of dimensions and arrangement of the essential devices for cashier's work. It means that a different layout can involve different tasks, so it is necessary to evaluate every single system in order to define the entity of required commitment for the worker. A supermarket checkout was considered in order to evaluate its workstation and to analyze the different tasks of two cashiers with similar anthropometric measurements. In particular one of them is affected by a widespread osteoarthritis, verified by instrumental tests and clinical examination. Several geometrical measures of the workstation were carried out while the cashiers were observed and filmed by videotapes during their activities. The study has been carried out by drawing the "preferred" and "maximal" working areas, as UNI EN ISO 14738:2004 suggests. Some tasks seem to be particularly critical especially for the cashier affected by osteoarthritis, who adopts compensatory postural mechanisms in order to use some devices which are not easy to reach inside the workstation, and which can induce pain to her. Finally the authors make some considerations about the opportunity of making a new layout of the workstation in order to find a more suitable arrangement of the devices.

BRCA1 and BRCA2 genetic testing in Italian breast and/or ovarian cancer families: mutation spectrum and prevalence and analysis of mutation prediction models.

BACKGROUND: Breast cancer is an extremely complex disease, characterized by a progressive multistep process caused by interactions of both genetic and non-genetic factors. A combination of BRCA1 and BRCA2 gene mutations appears responsible for about 20%-30% of the cases with breast cancer familial history. The prevalence of BRCA1/2 pathogenic mutations largely varies within different populations; in particular, the rate of mutations in Italian breast and/or ovarian cancer families is rather controversial and ranges from 8% to 37%. PATIENTS AND METHODS: Of the 152 breast/ovarian cancer families counseled in our centre, 99 were selected for BRCA1/2 mutation screening according to our minimal criteria. The entire coding sequences and each intron/exon boundary of BRCA1/2 genes were screened by direct sequencing (PTT limited to BRCA1 exon 11). For each proband, the a priori probability of carrying a pathogenic BRCA1/2 germline mutation was calculated by means of different mutation prediction models (BRCApro, IC and Myriad Table) in order to evaluate their performances. RESULTS: Our analysis resulted in the identification of 25 and 52 variants in the BRCA1 and BRCA2 genes, respectively. Seventeen of them represent novel variants, including four deleterious truncating mutations in the BRCA2 gene (472insA, E33X, C1630X and IVS6+1G>C). Twenty-seven of the 99 probands harbored BRCA1 (n = 15) and BRCA2 (n = 12) pathogenic germline mutations, indicating an overall detection rate of 27.3% and increasing by more than 15% the spectrum of mutations in the Italian population. Furthermore, we found the lowest detection rate (19.4%) in pure hereditary breast cancer family subset. All of the prediction models showed praises and faults, with the IC software being extremely sensitive but poorly specific, compared to BRCApro. In particular all models accumulated most false-negative prediction in the HBC subset. Interestingly preliminary results of a study addressing the presence of genomic rearrangements in HBC probands with BRCApro or IC prediction scores >/=95%, provided evidence for additional mutations undetectable with our conventional screening for point mutations. CONCLUSIONS: Altogether our results suggest that HBC families, the largest pool in our series, represent an heterogeneous group where the apparently faulty performances of the prediction models might be at least partially explained by the presence of additional kinds of BRCA1/2 alteration (such as genomic rearrangements) or by mutations on different breast cancer related genes.

Particulate matter cytotoxicity in cultured SH-SY5Y cells is modulated by simvastatin: Toxicological assessment for oxidative damage.

Epidemiological studies have shown a positive correlation between environmental particulate matter and adverse health effects. In particular, residual oil fly ash (ROFA) induces inflammation and reactive oxygen species (ROS), exerting not only local, but also systemic adverse effects. Previously, in an experimental animal model, we found that simvastatin (Sv) pretreatment was effective in preventing ROFA induced lung inflammation. Herein, using the human neuroblastoma SH-SY5Y cell line as a neurotoxicity in vitro model, we studied the potential Sv protective effect on ROFA cytotoxicity. We evaluated cell viability by the MTT assay, superoxide anion generation by NBT test, Nrf2 activation by immunofluorescence, apoptosis by cleaved-PARP and active-caspase 3 expressions, and senescence by ß-galactosidase activity. SH-SY5Y cells exposed to ROFA (10 and 50µg/ml) for 24h showed decreased cell viability, increased superoxide anion generation, apoptosis and senescence. Pretreatment with Sv (1µM) for 6 days, restored cell viability to basal levels, reduced ROFA-induced O2(-) generation as well as the number of apoptotic and senescent cells. Sv pretreatment stimulated the basal and ROFA-induced levels of Nrf2 nuclear translocation suggesting that activation of the cellular antioxidant defense system prevented particle-induced oxidative stress. In parallel, rescue experiments with mevalonate did not modify the effects of SV pretreatment in any of the parameters evaluated in this study. We conclude that simvastatin may provide neuroprotection against air particulate matter-induced neurotoxicity independently of its ability to inhibit cholesterol synthesis.