Cardiovascular changes, laboratory findings and pain scores in calves undergoing ultrasonography-guided bilateral rectus sheath block before herniorrhaphy: a prospective randomized clinical trial.

BACKGROUND: The study aimed to assess the clinical utility of a multiparametric approach to measure the impact of bilateral ultrasound-guided rectus sheath blocks (RSB) on heart rate, serum cortisol concentrations, and pain in calves undergoing herniorraphy. Fourteen calves were randomly assigned to receive either the RSB (RSB group, n = 7, injected with 0.3 mL/kg of bupivacaine 0.25% and 0.15 µg/kg of dexmedetomidine per side) or a sham injection (CG group, n = 7, injected with an equivalent volume of sterile saline solution). Monitoring included (i) continuous Holter recording from 120 min pre-surgery to 120 min post-surgery; (ii) serum cortisol concentration (SC) at -150 min pre-surgery (baseline), induction time, skin incision, end of surgical procedure (EP-t), and then 30 min, 45 min, 60 min, 120 min, 360 min after recovery; (iii) UNESP-Botucatu pain evaluation at -150 min pre-surgery and 30 min, 45 min, 60 min, 120 min, 240 min, 360 min after recovery. RESULTS: A significant difference in the heart rate was observed within the RSB group, in the time frame between 120 min to induction compared to the time frame between induction to EP-t period. The SC concentration was significantly higher in the CG at the skin incision. Calves in the RSB group recorded significantly lower pain scores at 45 min, 60 min, 120 and 240 min after recovery. CONCLUSIONS: The study demonstrated that monitoring heart rate and serum cortisol concentrations effectively quantified the effects of RSB during surgery. At the same time, the UNESP-Botucatu pain scale identified effects post-surgery when the calves regained consciousness. Overall, ultrasound-guided RSB appeared to enhance the well-being of calves undergoing herniorrhaphy.

HOME-BIO (sHOtgun MEtagenomic analysis of BIOlogical entities): a specific and comprehensive pipeline for metagenomic shotgun sequencing data analysis.

BACKGROUND: Next-Generation-Sequencing (NGS) enables detection of microorganisms present in biological and other matrices of various origin and nature, allowing not only the identification of known phyla and strains but also the discovery of novel ones. The large amount of metagenomic shotgun data produced by NGS require comprehensive and user-friendly pipelines for data analysis, that speed up the bioinformatics steps, relieving the users from the need to manually perform complex and time-consuming tasks. RESULTS: We describe here HOME-BIO (sHOtgun MEtagenomic analysis of BIOlogical entities), an exhaustive pipeline for metagenomics data analysis, comprising three independent analytical modules designed for an inclusive analysis of large NGS datasets. CONCLUSIONS: HOME-BIO is a powerful and easy-to-use tool that can be run also by users with limited computational expertise. It allows in-depth analyses by removing low-complexity/ problematic reads, integrating the analytical steps that lead to a comprehensive taxonomy profile of each sample by querying different source databases, and it is customizable according to specific users' needs.

Yes-Associated Protein 1 Is a Novel Calcium Sensing Receptor Target in Human Parathyroid Tumors.

The Hippo pathway is involved in human tumorigenesis and tissue repair. Here, we investigated the Hippo coactivator Yes-associated protein 1 (YAP1) and the kinase large tumor suppressor 1/2 (LATS1/2) in tumors of the parathyroid glands, which are almost invariably associated with primary hyperparathyroidism. Compared with normal parathyroid glands, parathyroid adenomas (PAds) and carcinomas show variably but reduced nuclear YAP1 expression. The kinase LATS1/2, which phosphorylates YAP1 thus promoting its degradation, was also variably reduced in PAds. Further, YAP1 silencing reduces the expression of the key parathyroid oncosuppressor multiple endocrine neoplasia type 1(MEN1), while MEN1 silencing increases YAP1 expression. Treatment of patient-derived PAds-primary cell cultures and Human embryonic kidney 293A (HEK293A) cells expressing the calcium-sensing receptor (CASR) with the CASR agonist R568 induces YAP1 nuclear accumulation. This effect was prevented by the incubation of the cells with RhoA/Rho-associated coiled-coil-containing protein kinase (ROCK) inhibitors Y27632 and H1152. Lastly, CASR activation increased the expression of the YAP1 gene targets CYR61, CTGF, and WNT5A, and this effect was blunted by YAP1 silencing. Concluding, here we provide preliminary evidence of the involvement of the Hippo pathway in human tumor parathyroid cells and of the existence of a CASR-ROCK-YAP1 axis. We propose a tumor suppressor role for YAP1 and LATS1/2 in parathyroid tumors.

Identification of differentially expressed microRNAs in the skin of experimentally sensitized naturally affected atopic beagles by next-generation sequencing.

Canine atopic dermatitis (AD) is a very common inflammatory skin disease, but limited data are available on the genetic characterization (somatic mutations, microarrays, and genome-wide association study (GWAS)) of skin lesions in affected dogs. microRNAs are good biomarkers in inflammatory and neoplastic diseases in people. The aim of this study was to evaluate microRNA expression in the skin of atopic beagles, before and after exposure to Dermatophagoides farinae. Four atopic and four unrelated age-matched healthy beagle dogs were enrolled. Total RNA was extracted from flash-frozen skin biopsies of healthy and atopic dogs. For the atopic dogs, skin biopsies were taken from non-lesional (day 0) and lesional skin (day 28 of weekly environmental challenge with Dermatophagoides farinae). Small RNA libraries were constructed and sequenced. The microRNA sequences were aligned to CanFam3.1 genome. Differential expressed microRNAs were selected on the basis of fold-change and statistical significance (fold-change ≥ 1.5 and p ≤ 0.05 as thresholds. A total of 277 microRNAs were sequenced. One hundred and twenty-one differentially regulated microRNAs were identified between non-lesional and healthy skin. Among these, two were increased amount and 119 were decreased amount. A total of 45 differentially regulated microRNAs between lesional and healthy skin were identified, 44 were decreased amount and one was increased amount. Finally, only two increased amount microRNAs were present in lesional skin when compared with that of non-lesional skin. This is the first study in which dysregulation of microRNAs has been associated with lesional and non-lesional canine AD. Larger studies are needed to understand the role of microRNA in canine AD.

Antibiotic dry buffalo therapy: effect of intramammary administration of benzathine cloxacillin against Staphylococcus aureus mastitis in dairy water buffalo.

BACKGROUND: Mastitis is one of the most costly diseases in Mediterranean buffalo (MB). At present, just a few specific antibiotics registered for this dairy specie have been synthetized. Efficacy of an antibiotic dry buffalo therapy (aDBT) against Staphylococcus aureus (S. aureus) mastitis, based on intra-quarter administration of 600 mg of benzathine cloxacillin, have been evaluated for the first time. Eighty MB's quarters received a drying-off therapy (aDBT-group) and 80 were left untreated (no-aDBT-group). They were sampled at drying-off (pre-treatment) and at the resumption of milking [< 10 days in milk (DIM)]. Fresh calver mastitis rate, dry period new mastitis rate, dry period cure rate, and persistent mastitis rate were calculated for clinical monitoring. Overall proportion of positive quarters/animals, quarters affected by mastitis or intramammary infections (IMI), effects on somatic cell count (SCC) and milk yield were also assessed. RESULTS: An inter-group difference (aDBT vs. no-aDBT) was recorded for all the indexes considered. An intra-group (drying-off vs. < 10 DIM) difference was detected in aDBT-group regarding the proportion of positive-cultured quarters and animals. Concerning the latter, an inter-groups difference was also recorded at second sampling. No clinical mastitis due to the S. aureus was observed. Regarding the subclinical ones, a higher intra-group difference was observed in aDBT than no-aDBT group, while an inter-group difference was recorded at second sampling. No protective effect was observed against IMI. SCC showed an inter-group difference at second sampling, while none difference was instead detected for milk yield. CONCLUSIONS: The effects against S. aureus mastitis of benzathine cloxacillin administration at drying-off were assessed for the first time in MB. Its use shows encouraging results in reducing the proportion of mastitis and positive animals at the resumption of the lactation.

Validation of a noseband pressure sensor algorithm as a tool for evaluation of feeding behaviour in dairy Mediterranean buffalo (Bubalus Bubalis).

This research communication addresses the goal of validating an algorithm to monitor natural occurrence of feeding behaviours in dairy Mediterranean buffalo based on the output of a noseband pressure sensor (RumiWatch®, halter). Several characteristics of the feeding behaviour were detected with a very high (ruminating boluses), high (chews per bolus) and moderate degree of correlation (chews per minute) with video analyses (gold standard). All of them were associated with a low mean difference with the gold standard, and the mean relative measurement error ranged between low (ruminating boluses) and moderate (chews per bolus and chews per minute). The proportion of correctly detected events for the variables rumination and eating time was 98 and 99%, respectively. The collection of data and subsequent evaluation of the parameters investigated may provide objective information on Mediterranean Buffalo behaviours allowing for reliable studies of the animal welfare in this ruminant in the future.

Efficacy of a polyvalent mastitis vaccine against Staphylococcus aureus on a dairy Mediterranean buffalo farm: results of two clinical field trials.

BACKGROUND: In the last years the knowledges on Mediterranean Buffalo (MB) mastitis is remarkably improving, nevertheless the attention has been never focused on vaccination as preventive strategy for the control of mastitis in these ruminates. The aim of the current study was to assess clinical efficacy over time of two different preventive vaccination protocols against S. aureus mastitis, in primiparous MB.Vaccinated (VG) and not-vaccinated (N-VG) groups, of 30 MB each one, were selected from two different herds (herd A: VG1 and N-VG1; herd B: VG2 and N-VG2) of the same farm. Herd A received a double vaccination (Startvac®, 45 and 10 days before calving, protocol A), while in herd B an additional administration was performed (52 days after calving, protocol B). Bacteriological milk culture and assessment of somatic cell count (SCC) were performed at 10, 30, 60 and 90 days in milk (DIM) from composite milk samples. After 90 DIM, daily milk yields and SCC values were monthly detected until dry-off. RESULTS: The overall incidence of positive MB for S. aureus was 40.8% (49/120) in VG1 and 43.3% (52/120) in N-VG1 (Protocol A), while 45.8% (55/120) and 50.8% (61/120) in VG2 and N-VG2 (Protocol B). The latter was associated with a significant decreased in prevalence (at 90 DIM) and incidence of mastitis (animals positive for S. aureus, SCC > 200^103, with or without clinical signs) in the vaccinated MB, while no difference occurred in protocol A. Moreover, herd B showed a significant reduction in prevalence of intramammary infection (animals positive for S. aureus, SCC < 200^103, no clinical signs) in the vaccinated MB at 60 DIM while no differences were detected in herd A, at any sampling time; N-VG2 had significantly higher overall SCC values than VG2 (4.97 ± 4.75 and 4.84 ± 4.60 Log10 cells/mL ± standard deviation, respectively), while no differences were recorded in herd A. CONCLUSIONS: The current investigation explores for the first time the clinical efficacy of vaccinations against S. aureus infections in MB, showing encouraging results regarding reduction in mastitis and somatic cell count; the polyvalent mastitis vaccine may be considered an additional tool for in-herd S aureus infection and should be associated to other control procedures to maximize its properties.

Validation of a pedometer algorithm as a tool for evaluation of locomotor behaviour in dairy Mediterranean buffalo.

This research communication validates an algorithm to monitor natural occurrence of locomotor behaviours in dairy Mediterranean buffalo based on the output of a 3-dimensional accelerometer (RumiWatch®, pedometer). Several characteristics of the locomotor behaviour were detected with a very high (up-right, lying and standing time) or high degree of correlation (walking time and number of strides) and a low mean difference with the video recording. The proportion of correctly detected events exceeded 99 % for the following variables: stand up and lie down events, as well as number of lying, standing or walking bouts. The mean relative measurement error was less than 10 % for the variables: lying, standing, up-right times and number of strides as compared with gold standard. This new algorithm may represent the base for a future early and real-time disease warning system aiming to gain higher health standard in these ruminants.

Assessment of foot health and animal welfare: clinical findings in 229 dairy Mediterranean Buffaloes (Bubalus bubalis) affected by foot disorders.

BACKGROUND: Lameness represents the third most important health-related cause of economic loss in the dairy industry after fertility and mastitis. Although, dairy Mediterranean Buffaloes (MB) and dairy cows share similar breeding systems predisposing to similar herd problems, published studies exploring its relevance and role in these ruminants are still rare and incomplete. The aims of this study were to describe the clinical findings of foot disorders (FDs) in dairy MB and their influence on animal welfare, determined by assessment of locomotion score (LS), body condition score (BCS) and cleanliness score (CS). RESULTS: Of 1297 multiparous MB submitted to routine trimming procedures, 229 buffaloes showed at least one FD. The prevalence of buffaloes affected by FDs was 17.7 %, while motility and lameness indexes were 84.1 % (1091/1297) and 15.9 % (206/1297), respectively. Overgrowth was present in 17.0 % (220/1297), corkscrew claw in 15.8 % (205/1297), interdigital phlegmon in 0.9 % (12/1297), white line abscess in 0.8 % (11/1297), digital dermatitis in 0.1 % (1/1297) and interdigital hyperplasia in 0.1 % (1/1297). Simultaneous presence of FDs was recorded in 17.0 % of MB (221/1297): overgrowth and corkscrew claw occurred together in 15.8 % of cases (205/1297), overgrowth and interdigital phlegmon in 0.3 % (4/1297), overgrowth and white line abscess in 0.8 % (11/1297), digital dermatitis and interdigital hyperplasia in 0.1 % (1/1297). The presence of FDs was always associated with lameness (LS > 2), except from 23 MB with simultaneous overgrowth and interdigital phlegmon occurrence. The majority of MB within the under-conditioned group (95.5 %, 43/45) and all those with CS > 2 (122/122) had a locomotion score above the threshold of normality (LS > 2). Furthermore, foot diseases such as interdigital hyperplasia, white line abscess and digital dermatitis or interdigital hyperplasia seemed to occur more frequently associated with decreased BCS and increased CS scores. CONCLUSIONS: This study describes for the first time the involvement of white line disease, interdigital phlegmona, digital dermatitis and interdigital hyperplasia in foot disorders of dairy Mediterranean buffalo and shows their association with an impairment of animal welfare.

Efficacy of pasireotide LAR for acromegaly: a prolonged real-world monocentric study.

Background: Acromegaly is caused by excessive growth hormone (GH) and insulin-like growth factor 1 (IGF1). Medical therapy plays a role as a treatment option for persistent disease after non-curative surgery or as a first-line therapy when surgery is not feasible. Pasireotide-LAR (Pas-LAR) is recommended for patients with acromegaly as second-line treatment. Aim: To evaluate the patients characteristics predictive of an adequate response to Pas-LAR and the long-term efficacy and safety of the Pas-LAR treatment. Methods: Data from 19 patients with active acromegaly, who were and resistant or intolerant to first-line medical therapy and were switched to pas-LAR have been retrospectively collected. We compared the baseline clinical and biochemical characteristics of patients who were found to respond to Pas-LAR therapy (responders, n=14) with those of patients who did not respond (non-responders, n=5). We then evaluated the Pas-LAR efficacy and safety during long-term follow-up in responders. Results: IGF1 normalization occurred in 71.4% of responders after one injection. IGF1 levels, [median(interquartile range) of the upper limit of the normal range (ULN) fold increase] were higher in non-responders compared to responders within the initial month of therapy [1.40(1.30-2.34) vs 0.70(0.55-1.25), respectively, p=0.009] and after three [1.77(1.74-2.29) vs 0.94(0.82-1.13), respectively, p=0.029] and six months [1.68(1.33-1.72) vs 1.00(0.65 -1.28), respectively, p=0.002]. Out of 6 patients with symptomatic headache (all in responder group), 5 and 1 reported the resolution and improvement of headache, respectively, already after the first injection. Median HbA1c levels tended to increase from baseline to 6 months both in responder (36 mMol/Mol to 42 mMol/Mol) and non-responder patients (45 mMol/Mol to 48 mMol/Mol). During long term follow up, in the responder group 2 new patients developed diabetes. Tumor shrinkage was observed in 6 out of 7 evaluated responders, with no cases of size increase during the long-term follow-up. Conclusion: Pas-LAR is effective and safe and the early identification of responders is possible just after the first administration.

From Adenoma to Carcinoma: the Unexpected Evolution of an Apparently Stable Adrenal Lesion.

BACKGROUND: There is a lack of solid long-term evidence with respect to the management over time of adrenal incidentalomas that miss clearly benign radiological features. We present the case of a 75-year-old man with a non-secreting adrenal mass, apparently stable in size (14 mm) and unchanged in features for 2 years, but subsequently diagnosed as adrenal carcinoma. CASE REPORT: The patient was referred to Grande Ospedale Metropolitano Niguarda in August 2022 due to the presence of a large lesion in the left adrenal site. In 2017, a 14 mm, 20 HU, round, regular-edged lesion was detected at a CT scan without contrast medium. Over the next two years, the patient was re-evaluated every 6 months with follow-up CT scans with no apparent densitometric or dimensional changes in the known lesion. In September 2022, 3 years after the last CT scan, the patient was hospitalised for pneumonia. An abdominal CT scan acquired during the hospitalisation showed an increase of the lesion to 14.5x10x12 cm. The patient subsequently underwent open nephrosurrenectomy, and histological examination confirmed the presence of an adrenal carcinoma (proliferation index 5%, Weiss score 7). No adjuvant therapy was administered, and the last CT scan in December 2022 was negative for the recurrence of the disease. CONCLUSION: Adrenal carcinoma usually presents as a clearly malignant lesion with rapid growth and a marked tendency to metastasise. This case highlights how an adrenal adenoma with indeterminate features is worthy of follow-up over time despite its apparent dimensional and radiological stability [1].

Factors related to longevity and mortality of dogs in Italy.

Besides its translational value, an improved understanding of dog longevity and mortality is necessary to guide health management decisions, breed selection, and improve dog welfare. In order to analyse the lifespan of dogs in Italy, identify the most common causes of death, and evaluate possible risk factors, anonymised medical records were collected from 9 veterinary teaching hospitals and 2 public health institutions. Data regarding breed, sex, neuter status, age, diagnosis, and mechanism of death were retrieved. Cause of death (COD) was classified by pathophysiologic process (PP) and organ system (OS). Of the 4957 dogs that died between 2004 and 2020 included in the study, 2920 (59.0%) were purebred, 2293 (46.2%) were female, 3005 (60.6%) were intact, 2883 (58.2%) were euthanised. Overall median longevity was 10.0 years. Median longevity was significantly longer for crossbreds, females, neutered dogs, and small-sized breeds. The breeds with the highest median age at death were the Yorkshire terrier, English cocker spaniel, West Highland white terrier, Italian volpino, and Shih Tzu, whilst the American bulldog, English bulldog, American pit bull terrier, Bernese mountain dog and the Maremma and the Abruzzes sheepdog had the lowest median age at death. The most frequent COD by PP was neoplasia (34.0%), which occurred more frequently in large breeds, namely German shepherd, Labrador retriever and Boxer. Degenerative diseases mostly affected small-sized dogs like Miniature pinscher and Dachshund. Regarding the OS involved, diseases of the renal/urinary system were most frequently responsible for COD (15.0%), prevalently degenerative and inflammatory/infectious. Substantial variation in median longevity according to causes of death by PP and OS was observed. These data are relevant for breeders, veterinary practitioners, and owners, to assist breed selection, facilitate early diagnosis, guide choice when purchasing a purebred dog and making health management decisions, and ultimately improve dog welfare.

Clinical efficacy of an ultrasound-guided bilateral rectus sheath block for umbilical hernia repair in calves: A prospective randomized trial.

Introduction: Surgical umbilical hernia repair is a frequent procedure in newborn calves, requiring mandatory pain management. This study aimed to develop an ultrasound-guided rectus sheath block (RSB) and to evaluate its clinical efficacy in calves undergoing umbilical herniorrhaphy under general field anesthesia. Methods: Gross and ultrasound anatomy of the ventral abdomen and the diffusion of a new methylene blue solution after injection within the rectus sheath were described in seven fresh calf cadavers. Then, fourteen calves undergoing elective herniorrhaphy were randomly assigned to receive either bilateral ultrasound-guided RSB with 0.3 mL/kg of bupivacaine 0.25% and 0.15 µg/kg of dexmedetomidine or 0.3 mL/kg of 0.9% NaCl (control). Intraoperative data included cardiopulmonary variables and anesthetic requirements. Postoperative data included pain scores, sedation scores and peri-incisional mechanical threshold assessed by force algometry at specific time points after anesthetic recovery. Treatments were compared using Wilcoxon rank-sum, Student's t-test, and Cox proportional hazard model as appropriate. Mixed effect linear models on rank, with random effect calf; fixed effects time, treatment, and their interaction were used to compare pain scores and mechanical thresholds over time. Significance was set at p = 0.05. Results and Discussion: Calves receiving RSB recorded lower pain scores between 45 - 120 minutes (p < 0.05) and at 240 min after recovery (p = 0.02). And they recorded higher mechanical thresholds between 45 and 120 min after surgery (p < 0.05). Ultrasound-guided RSB provided effective perioperative analgesia in calves undergoing herniorrhaphy under field conditions.

Efficacy of oral, topical and extended-release injectable formulations of moxidectin combined with doxycycline in Dirofilaria immitis naturally infected dogs.

BACKGROUND: Several studies in both experimentally and naturally infected dogs have reported the adulticide effect of a combination of macrocyclic lactones and doxycycline against Dirofilaria immitis, showing that these protocols can be used as an alternative to melarsomine. The present study evaluated the efficacy of oral, topical and extended-release injectable formulations of moxidectin when combined with doxycycline in dogs naturally infected with D. immitis from a shelter located in southern Italy. METHODS: Thirty dogs with naturally acquired D. immitis infection were divided in three groups (G) and treated with oral moxidectin (G1) once a month for 9 consecutive months, topical moxidectin (G2) once a month for 9 consecutive months or extended release moxidectin injectable (G3) at enrolment and again at 6 months (Day 180). All treatment groups received doxycycline for the first 30 days. Microfilarial concentrations in 1 ml (mff/ml) blood were determined monthly for 9 months with the modified Knott's test. A clinical scoring system was employed for each dog enrolled in the study based on thoracic radiography and cardiac ultrasound (CU) examinations performed at Day - 15 (before treatment) and at Day 180. RESULTS: Results from the present study suggest that the majority of dogs from all treatment groups became antigen negative, as evaluated at Day 270: 9/10 dogs (90.0%) from G1, 6/10 dogs (60.0%) from G2 and 8/10 dogs (80.0%) from G3. Improvement of radiographic alterations was observed in all treatment groups, and almost all dogs were cleared of pulmonary abnormalities by 6 months from the beginning of treatment (P = 0.000). Cardiac ultrasound examination showed a progressive improvement of cardiac function in a limited number of animals (4/30). CONCLUSIONS: The combination of doxycycline and three different formulations of moxidectin leads to antigen-negative status in naturally infected dogs.

Metabolic Changes Associated with Different Levels of Energy Deficits in Mediterranean Buffaloes during the Early Lactation Stage: Type and Role of the Main Lipid Fractions Involved.

Cell function and energy redistribution are influenced by lipid classes (phospholipids (PLs), free fatty acids (FFAs), triglycerides (TGs), and cholesterol esters (CEs)). The aim of this study was to investigate metabolic alterations that are related to changes in lipid classes according to different levels of energy deficits in early lactating Mediterranean buffaloes (MBs). Sixty-three MBs were enrolled at the beginning of lactation using an observational study with a cross-sectional experimental design. Serum ß-hydroxybutyrate (BHB) levels were used to group the animals into a healthy group (Group H; n = 38; BHB < 0.70 mmol/L) and hyperketonemia risk group (Group K; n = 25; BHB ≥ 0.70 mmol/L). Statistical analysis was performed using a linear model that included the effect of the group and body condition score to assess differences in fatty acid (FA) concentrations. A total of 40 plasma FAs were assessed in each lipid class. Among the FAs, eight PLs, seven FFAs, four TGs, and four CEs increased according to BHB levels, while three FFAs, three TGs, and one CE decreased. The changes among lipid class profiles suggested the influence of inflammatory response, liver metabolism, and the state of body lipid reserves. In addition, the possible similarities of buffaloes at risk of hyperketonemia with ketotic cows suggest the necessity of further investigations in these ruminants.

Osteocalcin modulates parathyroid cell function in human parathyroid tumors.

Introduction: The bone matrix protein osteocalcin (OC), secreted by osteoblasts, displays endocrine effects. We tested the hypothesis that OC modulates parathyroid tumor cell function. Methods: Primary cell cultures derived from parathyroid adenomas (PAds) and HEK293 cells transiently transfected with the putative OC receptor GPRC6A or the calcium sensing receptor (CASR) were used as experimental models to investigate γ-carboxylated OC (GlaOC) or uncarboxylated OC (GluOC) modulation of intracellular signaling. Results: In primary cell cultures derived from PAds, incubation with GlaOC or GluOC modulated intracellular signaling, inhibiting pERK/ERK and increasing active ß-catenin levels. GlaOC increased the expression of PTH, CCND1 and CASR, and reduced CDKN1B/p27 and TP73. GluOC stimulated transcription of PTH, and inhibited MEN1 expression. Moreover, GlaOC and GluOC reduced staurosporin-induced caspase 3/7 activity. The putative OC receptor GPRC6A was detected in normal and tumor parathyroids at membrane or cytoplasmic level in cells scattered throughout the parenchyma. In PAds, the membrane expression levels of GPRC6A and its closest homolog CASR positively correlated; GPRC6A protein levels positively correlated with circulating ionized and total calcium, and PTH levels of the patients harboring the analyzed PAds. Using HEK293A transiently transfected with either GPRC6A or CASR, and PAds-derived cells silenced for CASR, we showed that GlaOC and GluOC modulated pERK/ERK and active ß-catenin mainly through CASR activation. Conclusion: Parathyroid gland emerges as a novel target of the bone secreted hormone osteocalcin, which may modulate tumor parathyroid CASR sensitivity and parathyroid cell apoptosis.

Spontaneous recovery of bone mass after cure of endogenous hypercortisolism.

Patients with Cushing's syndrome (CS) develop osteopenia-osteoporosis. The present study evaluates the recovery of bone mass within 2 years after remission of hypercortisolism and in long term follow up, an issue rarely addressed. Twenty patients (6M, 14F, 3 post-menopausal, 15-64 years old), 15 with Cushing's disease, 2 with ectopic ACTH syndrome, 3 with ACTH-independent CS were studied. BMD, T and Z scores at lumbar spine and proximal femur were assessed by dual-energy X-ray absorptiometry before and 7-33 months after treatment of hypercortisolism. Five patients were treated with bisphosphonates. Four patients had hypogonadism and 4 GH-deficiency. At baseline all patients showed osteopenia/osteoporosis and the spine appeared more damaged than the femur; femur BMD was positively related with body mass index (BMI). No correlations were observed between spine and femur bone parameters and duration of disease or severity of hypercortisolism. Bone parameters did not differ in patients with or without GH or other pituitary deficiencies. After cure of hypercortisolism a significant improvement in spine BMD, Z and T scores and in femur Z and T scores was observed with normalization in 3 patients; there was no significant difference in percent improvement between femur and spine. The increase in bone parameters at spine and femur was independent from values at baseline. The percent increase in spine T and Z scores was positively related with time elapsed since cure. Bisphosphonates did not influence the recovery of bone mineralization. In long term follow up, after a median period of 7 years a further improvement in bone density was observed in 100% of patients at spine and in 9/11 at femur, although 8/11 patients still had femoral and/or vertebral T score in the range of osteopenia/osteoporosis. Spontaneous improvement of osteoporosis after cure of hypercortisolism occurs both at spine and femur, is independent from basal conditions and not affected by bisphosphonates. The improvement at spine depends on time since cure.

The Use of Virtual-Problem-Based Learning during COVID-19 Pandemic Emergency: Veterinary Students' Perception.

The COVID-19 global pandemic emergency forced us to replace the "traditional and in presence" clinical, pre-graduating, veterinary medical training with clinical virtual-problem-based learning (v-PBL). This prospective cross-sectional case-control study aimed to evaluate the students' perception of the v-PBLs compared to the traditional veterinary clinical training (t-VCT). The t-VCT consisted of supervised management of clinical cases admitted at the Veterinary Teaching Hospital and in the field. The v-PBL consisted of genuine clinical cases shared by tutors throughout an online platform. A survey was delivered to all the fifth-year students who completed the t-VCT or the v-PBL. The survey was completed by 49% of the students. Overall student satisfaction regarding the training experiences was high in both groups, but it was less in the v-PBL than in the t-VCT group. The students of the v-PBL group perceived that they could not improve their practical clinical skills through online sessions, and they emphasized how it could be employed as support for traditional practical activities. All the students are satisfied with the supervision and considered the training correctly focused on relevant learning objectives and the task clearly explained. Stimulating the integration of knowledge and lifelong learning skills replicating life experiences the v-PBLs represented an attractive curricular alternative for veterinary education.

Assessment of feeding, ruminating and locomotion behaviors in dairy cows around calving - a retrospective clinical study to early detect spontaneous disease appearance.

The study aims to verify the usefulness of new intervals-based algorithms for clinical interpretation of animal behavior in dairy cows around calving period. Thirteen activities associated with feeding-ruminating-locomotion-behaviors of 42 adult Holstein-Friesian cows were continuously monitored for the week (wk) -2, wk -1 and wk +1 relative to calving (overall 30'340 min/animal). Soon after, animals were retrospectively assigned to group-S (at least one spontaneous diseases; n = 24) and group-H (healthy; n = 18). The average activities performed by the groups, recorded by RumiWatch® halter and pedometer, were compared at the different weekly intervals. The average activities on the day of clinical diagnosis (dd0), as well as one (dd-1) and two days before (dd-2) were also assessed. Differences of dd0 vs. dd-1 (ΔD1), dd0 vs. wk -1 (ΔD2), and wk +1 vs. wk -1 (Δweeks) were calculated. Variables showing significant differences between the groups were used for a univariate logistic regression, a receiver operating characteristic analysis, and a multivariate logistic regression model. At wk +1 and dd0, eating- and ruminating-time, eating- and ruminate-chews and ruminating boluses were significantly lower in group-S as compared to group-H, while other activity time was higher. For ΔD2 and Δweeks, the differences of eating- and ruminating-time, as well as of eating-and ruminate-chews were significantly lower in group-S as compared to group-H. Concerning the locomotion behaviors, the lying time was significantly higher in group-S vs. group-H at wk +1 and dd-2. The number of strides was significantly lower in group-S compared to group-H at wk +1. The model including eating-chews, ruminate-chews and other activity time reached the highest accuracy in detecting sick cows in wk +1 (area under the curve: 81%; sensitivity: 73.7%; specificity: 82.4%). Some of the new algorithms for the clinical interpretation of cow behaviour as described in this study may contribute to monitoring animals' health around calving.

Adrenal schwannoma: why should endocrinologists be aware of this uncommon tumour?

PURPOSE: Adrenal schwannomas (AS) are rare tumours arising from Schwann cells. Due to the high prevalence of adrenal incidentalomas, running into very rare adrenal tumours has become a possibility for high volume expert centres. So far, the clinical behaviour and the radiological characteristics of AS make the pre-operative diagnosis of AS extremely challenging. Due to limited information available, we wanted to summarise the main features of this tumours, in order to raise the profile of an uncommon disease. METHODS: We performed a MEDLINE and EMBASE research to review the literature. We found 57 case reports and case series and a total of 169 cases, including 2 more cases found in our Institutions. We collected data regarding year of publication, sex, age, and, when available, clinical presentation, hormonal data, radiological features, tumour site and size, treatment, histology and follow-up. RESULTS: We analysed and discussed the clinical, radiological and pathological characteristics of cases identified, underlying the critical aspects of assessment and management of these tumours which still remain questioned, as, currently, pathologic examination is the only way to make the diagnosis. CONCLUSIONS: The pre-operative diagnosis of AS is more than challenging and pathologic examination is so far the only way to make a certain diagnosis. Therefore, it is important to consider also the AS in the list of possible diagnoses when faced with a large not secreting adrenal tumour, with suspicious radiological features.