Valores de digestibilidade e composição química e bromatológica de alguns alimentos para suínos.

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Identification of key receptor residues discriminating human chorionic gonadotropin (Hcg)-and luteinizing hormone (lh)-specific signaling

(1) The human luteinizing hormone (LH)/chorionic gonadotropin (hCG) receptor (LHCGR) discriminates its two hormone ligands and differs from the murine receptor (Lhr) in amino acid residues potentially involved in qualitative discerning of LH and hCG. The latter gon-adotropin is absent in rodents. The aim of the study is to identify LHCGR residues involved in hCG/LH discrimination. (2) Eight LHCGR cDNAs were developed, carrying “murinizing” mutations on aminoacidic residues assumed to interact specifically with LH, hCG, or both. HEK293 cells expressing a mutant or the wild type receptor were treated with LH or hCG and the kinetics of cyclic adenosine monophosphate (cAMP) and phosphorylated extracellular signal-regulated ki-nases 1/2 (pERK1/2) activation was analyzed by bioluminescence resonance energy transfer (BRET). (3) Mutations falling within the receptor leucine reach repeat 9 and 10 (LRR9 and LRR10; K225S +T226I and R247T), of the large extracellular binding domain, are linked to loss of hormone-specific induced cAMP increase, as well as hCG-specific pERK1/2 activation, leading to a Lhr-like modulation of the LHCGR-mediated intracellular signaling pattern. These results support the hypothesis that LHCGR LRR domain is the interaction site of the hormone β-L2 loop, which differs between LH and hCG, and might be fundamental for inducing gonadotropin-specific signals. (4) Taken to-gether, these data identify LHCGR key residues likely evolved in the human to discriminate LH/hCG specific binding

Anestesia locoregionale dell'orecchio nel cavallo: descrizione del blocco tramite l'ausilio di un neurostimolatore

Introduzione. L\u2019esecuzione di procedure diagnostiche e terapeutiche a carico dell\u2019orecchio del ca- vallo \ue8 spesso complicata e frustrante per il veterinario. Ci\uf2 \ue8 particolarmente vero per procedure dolorose quali biopsie o gestione di ferite che richiedono l\u2019immobilit\ue0 del paziente1. A tale propo- sito nell\u2019ambito chirurgico, di grossi e piccoli animali, si \ue8 assistito negli ultimi anni, ad un cre- scente interesse nei confronti di tecniche di analgesia selettiva dell\u2019orecchio. Il blocco diretto di ner- vi che innervano la pinna a livello della base dell\u2019orecchio costituiscono una tecnica in grado di ri- durre lo stress del paziente e il tempo richiesto per l\u2019esecuzione delle procedure1. Nonostante un\u2019ot- tima conoscenza anatomica locale, la corretta localizzazione nervosa viene meno a causa di un\u2019ele- vata variabilit\ue0 individuale1. La stimolazione nervosa periferica (tramite neurostimolatore) permet- te di applicare in modo mirato ed in stretta vicinanza alla fibra nervosa, una serie di stimoli elettri- ci che, provocando una contrazione muscolare, consentono di identificare le strutture di interesse. L\u2019utilizzo del neurostimolatore consente una veloce e precisa localizzazione nervosa e abbassa i ri- schi di lesioni da inoculazione intraneurale. Descrizione del caso. Un cavallo di 9 anni, di 420 Kg viene riferito per drenaggio chirurgico di un otoematoma a carico dello scafo dell\u2019orecchio sinistro. L\u2019innervazione auricolare si compone di 3 nervi fondamentali: branca mandibolare del nervo trigemino, branca auricolopalpebrale del nervo facciale e secondo nervo cervicale o grande auricolare. Si decide quindi per il blocco di questi 3 ner- vi tramite l\u2019utilizzo di un neurostimolatore. Il soggetto viene sedato e lo strumento (Vygon Italia) viene connesso ad un ago atraumatico (22G, 50 mm, 30\ub0), infisso nel punto di repere specifico per i singoli nervi e viene impostato ad una intensit\ue0 di 1.0 mA con una stimolazione di 2 Hz della du- rata di 0.1 ms, per ottenere una contrazione dei muscoli target2. Per anestetizzare la branca mandi- bolare del nervo trigemino l\u2019ago, dopo accurata tricotomia e preparazione asettica della cute del- l\u2019area, viene infisso a livello dell\u2019articolazione temporo-mandibolare con direzione rostro-caudale e latero-mediale fino ad ottenere una contrazione dei muscoli digastrico, pterigoideo mediale e la- terale e massetere (spostamento rostrale della pinna e masticazione). Vengono quindi inoculati 5 ml di mepivacaina cloridrato 2%. Viene poi individuata ed anestetizzata la branca auricolo-palpebrale del nervo facciale. Il suo punto di repere \ue8 rappresentato dalla cresta occipitale e dalla base della pinna, costituita dai margini mediale e laterale dell\u2019elice. Dopo accurata tricotomia e preparazione asettica della cute dell\u2019area, l\u2019ago atraumatico viene inserito al centro del triangolo idealmente di- segnato dai due vertici tra la base della pinna e la cresta occipitale. Con le stesse procedure utiliz- zate precedentemente viene eseguita una stimolazione dei muscoli della pinna e della palpebra su- periore e, una volta ottenuti movimenti auricolari caudorostrali e ammiccamento, vengono inocula- ti 5 ml di mepivacaina cloridrato 2%. Per effettuare il blocco del II nervo cervicale, l\u2019ago atraumatico viene inserito ventrocaudalmente all\u2019ala dell\u2019atlante in direzione lateromediale e leggermente ventrodorsale sino ad ottenere una con- trazione dei muscoli del collo e movimenti di abduzione dell\u2019orecchio. Vengono quindi inoculati 5 ml di mepivacaina cloridrato 2%. Dopo 10 minuti dall\u2019esecuzione di ogni singolo blocco viene testata la sensibilit\ue0 tattile e dolorifi- ca dell\u2019orecchio e, visto l\u2019esito favorevole, si procede all\u2019intervento chirurgico. Conclusioni. Il blocco della branca mandibolare del trigemino, della branca auricolopalpebrale del nervo facciale e della branca auricolare del II nervo cervicale \ue8 risultato di facile esecuzione ed ef- ficace per la desensibilizzazione dell\u2019orecchio in tutta la sua struttura. La tecnica \ue8 stata ben tolle- rata e ha permesso di eseguire in maniera agevole la chirurgia riducendo il dosaggio dei farmaci anestetici utilizzati per l\u2019anestesia generale. Questo tipo di anestesia locoregionale potrebbe essere efficace per effettuare procedure chirurgiche a carico dell\u2019orecchio con la sola sedazione dell\u2019ani- male (drenaggio otoematoma, escissione masse neoplastiche o criochirurgia), ovviando cos\uec ai ri- schi legati all\u2019anestesia generale3. La buona conoscenza anatomica e l\u2019utilizzo dello stimolatore nervoso periferico ha permesso la cor- retta localizzazione nervosa con conseguente desensibilizzazione del sito di chirurgia. L\u2019impiego di anestetici locali a media/lunga durata d\u2019azione consente una riduzione del dolore nel periodo pe- rioperatorio, condizione indispensabile affinch\ue9 diminuisca il rischio di autotraumatismi. In ultima analisi, il blocco dei nervi sopracitati ha permesso non solo l\u2019esecuzione della procedura chirurgica ma anche una miglior gestione del paziente nel periodo post-operatorio

Polygenic Susceptibility to Papillary Thyroid Cancer in Italian Subjects.

olygenic Susceptibility to Papillary Thyroid Cancer in Italian Subjects INTRODUCTION AND AIM. Thyroid cancer is the most common endocrine neoplasia, with an estimated age- standardized incidence rate of 6.7 per 100000 worldwide in 2018 [1]. This rate is rapidly increasing and papillary thy- roid carcinoma (PTC) is the main histotype. PTC suscepti- bility is the result of genetic predisposition, environmental factors and lifestyle. We studied the genetic combination that characterizes PTC affected subjects, differentiating them from healthy controls. METHODS AND RESULTS. We considered the genetic variants (SNPs) significantly associated with PTC on the PubMed database. 184 informative SNPs were selected, considering linkage disequilibrium. Then, SNPs data were extracted from the online 1000 Genomes database,comprising genome of 2504 unselected individuals col- lected worldwide. The combination of 184 SNPs associ- ated with PTC was used to group individuals in different risk-clusters according to their genetic structure, calcu- lated by Bayesian statistics, as previously performed for polycystic ovary syndrome [2]. Individuals were distrib- uted among 7 groups worldwide, indicating different de- gree of genetic predisposition to PTC. We then considered genetic data from about 1200 individuals (697 PTC versus 497 healthy controls) of Central/South Italian origin reg- istered in a GWAS, specific for PTC [3]. This first analysis was refined using the 33 SNPs reasonably most causa- tive of genetic clustering (26 with p<0.05 at trend test in GWAS and 7 with p<0.05 in the model of recessive inher- itance). At multivariate logistic regression analysis, PTC and healthy controls resulted genetically different (ODDS RATIO 188.6, 95%CI 64.35-552.8), revealing diverse pre- disposition to develop cancer. Afterwards, these results have been confirmed in an independent cohort of Italian subjects (234 PTC and 100 controls). Then, the genetic structure of each subject was indicated as a percentage of affinity to each risk-cluster and re-analyzed together with other risk factors: sex, body-mass index, area of origin and familiarity (quantified in a growing score as the degree of kinship increases). These data were analyzed together by principal component analysis and clustering of the two groups was even more pronounced. The most contributive factors to the diversity between PTC and healthy controls were genetics and familiarity. CONCLUSION. We demonstrated that PTC affected subjects are genetically different from healthy controls, and that the difference is identifiable in a peculiar combi- nation of genetic variants

Sphingosine-1 phosphate induces cAMP/PKA-independent phosphorylation of the cAMP response element-binding protein (CREB) in granulosa cells

Background and aims: Sphingosine-1 phosphate (S1P) is a lysosphingolipid present in the ovarian follicular fluid. The role of the lysosphingolipid in gonads of the female is widely unclear. At nanomolar concentrations, S1P binds and activates five specific G protein-coupled receptors (GPCRs), known as S1P1-5, modulating different signaling pathways. S1P1 and S1P3 are highly expressed in human primary granulosa lutein cells (hGLC), as well as in the immortalized human primary granulosa cell line hGL5. In this study, we evaluated the signaling cascade activated by S1P and its synthetic analogues in hGLC and hGL5 cells, exploring the biological relevance of S1PR-stimulation in this context. METHODS AND RESULTS. hGLC and hGL5 cells were treated with a fixed dose (0.1 \u3bcM) of S1P, or by S1P1- and S1P3-specific agonists SEW2871 and CYM5541. In granulosa cells, S1P and, at a lesser extent, SEW2871 and CYM5541, potently induced CREB phosphorylation. No cAMP production was detected and pCREB activation occurred even in the presence of the PKA inhibitor H-89. Moreover, S1P-dependent CREB phosphorylation was dampened by the mitogen-activate protein kinase (MEK) inhibitor U0126 and by the L-type Ca2+ channel blocker verapamil. The complete inhibition of CREB phosphorylation occurred by blocking either S1P2 or S1P3 with the specific receptor antagonists JTE-013 and TY52156, or under PLC/PI3K depletion. S1P-dependent CREB phosphorylation induced FOXO1 and the EGF-like epiregulin-encoding gene (EREG), confirming the exclusive role of gonadotropins and interleukins in this process, but did not affect steroidogenesis. However, S1P or agonists did not modulate granulosa cell viability and proliferation in our conditions. Conclusions: This study demonstrates for the first time that S1P may induce a cAMP-independent activation of pCREB in granulosa cells, although this is not sufficient to induce intracellular steroidogenic signals and progesterone synthesis. S1P-induced FOXO1 and EREG gene expression suggests that the activation of S1P\u2013S1PR axis may cooperate with gonadotropins in modulating follicle development

Genetic Markers Associated to Dyslipidemia in HIV-Infected Individuals on HAART

This study evaluated the impact of 9 single nucleotide polymorphisms (SNPs) in 6 candidate genes (APOB, APOA5, APOE, APOC3, SCAP, and LDLR) over dyslipidemia in HIV-infected patients on stable antiretroviral therapy (ART) with undetectable viral loads. Blood samples were collected from 614 patients at reference services in the cities of Porto Alegre, Pelotas, and Rio Grande in Brazil. The SNPs were genotyped by conventional polymerase chain reaction (PCR) and real-time PCR. The prevalence of dyslipidemia was particularly high among the protease inhibitors-treated patients (79%). APOE (rs429358 and rs7412) genotypes and APOA5 −1131T>C (rs662799) were associated with plasma triglycerides (TG) and low-density-lipoprotein cholesterol levels (LDL-C). The APOA5 −1131T>C (rs662799) and SCAP 2386A>G (rs12487736) polymorphisms were significantly associated with high-density-lipoprotein cholesterol levels. The mean values of the total cholesterol and LDL-C levels were associated with both the APOB SP Ins/Del (rs17240441) and APOB XbaI (rs693) polymorphisms. In conclusion, our data support the importance of genetic factors in the determination of lipid levels in HIV-infected individuals. Due to the relatively high number of carriers of these risk variants, studies to verify treatment implications of genotyping before HAART initiation may be advisable to guide the selection of an appropriate antiretroviral therapy regimen

Preservation of Axillary Lymph Nodes Compared with Complete Dissection in T1–2 Breast Cancer Patients Presenting One or Two Metastatic Sentinel Lymph Nodes: The SINODAR-ONE Multicenter Randomized Clinical Trial

Background: The SINODAR-ONE trial is a prospective noninferiority multicenter randomized study aimed at assessing the role of axillary lymph node dissection (ALND) in patients undergoing either breast-conserving surgery or mastectomy for T1–2 breast cancer (BC) and presenting one or two macrometastatic sentinel lymph nodes (SLNs). The endpoints were to evaluate whether SLN biopsy (SLNB) only was associated with worsening of the prognosis compared with ALND in terms of overall survival (OS) and relapse. Methods: Patients were randomly assigned (1:1 ratio) to either removal of ≥ 10 axillary level I/II non-SLNs followed by adjuvant therapy (standard arm) or no further axillary treatment (experimental arm). Results: The trial started in April 2015 and ceased in April 2020, involving 889 patients. Median follow-up was 34.0 months. There were eight deaths (ALND, 4; SNLB only, 4), with 5-year cumulative mortality of 5.8% and 2.1% in the standard and experimental arm, respectively (p = 0.984). There were 26 recurrences (ALND 11; SNLB only, 15), with 5-year cumulative incidence of recurrence of 6.9% and 3.3% in the standard and experimental arm, respectively (p = 0.444). Only one axillary lymph node recurrence was observed in each arm. The 5-year OS rates were 98.9% and 98.8%, in the ALND and SNLB-only arm, respectively (p = 0.936). Conclusions: The 3-year survival and relapse rates of T1–2 BC patients with one or two macrometastatic SLNs treated with SLNB only, and adjuvant therapy, were not inferior to those of patients treated with ALND. These results do not support the use of routine ALND