Combining germline, tissue and liquid biopsy analysis by comprehensive genomic profiling to improve the yield of actionable variants in a real-world cancer cohort.

BACKGROUND: Comprehensive next-generation sequencing is widely used for precision oncology and precision prevention approaches. We aimed to determine the yield of actionable gene variants, the capacity to uncover hereditary predisposition and liquid biopsy appropriateness instead of, or in addition to, tumor tissue analysis, in a real-world cohort of cancer patients, who may benefit the most from comprehensive genomic profiling. METHODS: Seventy-eight matched germline/tumor tissue/liquid biopsy DNA and RNA samples were profiled using the Hereditary Cancer Panel (germline) and the TruSight Oncology 500 panel (tumor tissue/cfDNA) from 23 patients consecutively enrolled at our center according to at least one of the following criteria: no available therapeutic options; long responding patients potentially fit for other therapies; rare tumor; suspected hereditary cancer; primary cancer with high metastatic potential; tumor of unknown primary origin. Variants were annotated for OncoKB and AMP/ASCO/CAP classification. RESULTS: The overall yield of actionable somatic and germline variants was 57% (13/23 patients), and 43.5%, excluding variants previously identified by somatic or germline routine testing. The accuracy of tumor/cfDNA germline-focused analysis was demonstrated by overlapping results of germline testing. Five germline variants in BRCA1, VHL, CHEK1, ATM genes would have been missed without extended genomic profiling. A previously undetected BRAF p.V600E mutation was emblematic of the clinical utility of this approach in a patient with a liver undifferentiated embryonal sarcoma responsive to BRAF/MEK inhibition. CONCLUSIONS: Our study confirms the clinical relevance of performing extended parallel tumor DNA and cfDNA testing to broaden therapeutic options, to longitudinally monitor cfDNA during patient treatment, and to uncover possible hereditary predisposition following tumor sequencing in patient care.

Comparison of the distraction index and Norberg angle with radiographic grading of canine hip dysplasia.

The Orthopedic Foundation for Animals (OFA) radiographic grading system stands as a well-established benchmark for categorizing hip dysplasia in canines. Although more objective techniques to quantify early hip laxity in young dogs, such as the Norberg angle (NA) and distraction index (DI), have been documented, there is a scarcity of published studies that directly compare these radiographic measures. The primary objectives of this prospective, analytical study were to assess and compare the NA and DI measures with OFA grades within a cohort of dogs. Thirty dogs (57 hip joints) were evaluated, with a mean age of 5.4 ± 2.9 years and a mean body weight of 34.3 ± 7.1 kg. The discriminative power of hip dysplasia for DI and NA used OFA as a reference were compared by multivariate discriminant analysis test. Using the DI (≤0.3) 12 joints were classified as normal, using NA (≥105°) 24 joints and using OFA (≤3) 29. When comparing the discriminant power of NA with OFA resulting similar (P = .8324) with correct classifications of 81% using cross validation, however, the discriminant power of dysplasia of the DI resulted significantly greater (P = .0034) than OFA with correct classifications of only 59% using cross-validation, consequently it was possible determined that of 29 patients with OFA ≤ 3, 20 had DI > 0.3. These findings demonstrated that phenotypic healthy dogs according to OFA grading can present passive hip joint laxity, as demonstrated by radiographic evaluation with distraction. Furthermore, DI can contribute to the screening of CHD when evaluated in conjunction with OFA grading.

Early Surgical Management of Medial Patellar Luxation in Juvenile Dogs.

The aim of this study was to analyse a series of five cases of early surgical treatment in juvenile canine patients with high-grade medial patellar luxation. Canine patients, with immature skeleton, affected by grade III or IV medial patellar luxation and treated by multimodal surgical techniques were selected. Five dogs of different breeds, mean age 7.2 ± 3.0 months, met the inclusion criteria. Simultaneous distal femoral varus and external tibial torsion were identified in three of the five cases, whereas in the others, femoral varus with associated tibial valgus and isolated external tibial torsion were observed. Four of the five patients were treated surgically with closing wedge osteotomy of the distal femur, and three of them underwent tibial tuberosity transposition simultaneously. Only two animals required trochleoplasty. Mean time to beginning of weight bearing was 9.8 ± 5.5 days, whereas time to bone healing was 55 ± 24 days. Bone realignment of the extensor apparatus and return to limb support were achieved in all cases. Postoperative reintervention was needed in one patient; however, it was not linked to the initial surgery. Patients were followed up to 1 year postoperatively, with maintenance of extensor alignment and no late complications. The early surgical approach has been shown to be effective in the definitive treatment of juvenile dogs affected by high-grade medial patellar luxation.

Oxygen transfer and gas holdup in airlift bioreactors assembled with helical flow promoters.

Bioreactors can perform biochemical conversions mediated by biocatalysts, such as enzymes, animal cells, plants, and microorganisms. Among several existing models, airlift bioreactors are devices with the low shear environment and good mass transfer with low energy consumption, employed in several biochemical processes. The fluid flow is enabled through air injection by the sparger located at the bioreactor base. Despite its simple geometry compared with the conventional bioreactors, airlift performance can be optimized via geometrical modifications. Therefore, the objective of this work was to evaluate the effects of the addition of helical flow promoters, positioned in the riser and/or downcomer regions of an airlift of concentric tubes measuring the volumetric oxygen coefficient (kLa) and gas holdup. The results obtained by varying the gas flow rate from 1.0 to 4.0 vvm allowed the system evaluation of oxygen transfer and gas holdup. The inclusion of helical flow promoters increased the kLa, reaching up to 23% in oxygen transfer compared to tests without helicoids and up to 14% increase in the gas holdup. The inclusion of helical flow promotors was beneficial for all gas flow rates. Thus, including these flow promoters is an effective strategy to increase the oxygen transfer rate for bioprocess optimization.

Effects of Human Observer Presence on Pain Assessment Using Facial Expressions in Rabbits.

The goal of this study was to evaluate the effect of a human observer on Rabbit Grimace Scale (RbtGS) scores. The study scored video footage taken of 28 rabbits before and after orthopedic surgery, as follows: 24 h before surgery ( baseline), 1 h after surgery ( pain), 3 h after analgesia administration ( analgesia), and 24 h after surgery ( 24h) in the presence and absence of an observer. Videos were assessed twice in random order by 3 evaluators who were blind to the collection time and the presence or absence of an observer. Responses to pain and analgesia were evaluated by comparing the 4 time points using the Friedman test, followed by the Dunn test. The influence of the presence or absence of the observer at each time point was evaluated using the Wilcoxon test. Intra- and interrater reliabilities were estimated using the intraclass correlation coefficient. The scale was responsive to pain, as the scores increased after surgery and had decreased by 24 h after surgery. The presence of the observer reduced significantly the RbtGS scores (median and range) at pain (present, 0.75, 0 to 1.75; absent, 1, 0 to 2) and increased the scores at baseline (present, 0.2, 0 to 2; absent, 0, 0 to 2) and 24h after surgery (present, 0.33, 0 to 1.75; absent, 0.2, 0 to 1.5). The intrarater reliability was good (0.69) to very good (0.82) and interrater reliability was moderate (0.49) to good (0.67). Thus, the RbtGS appeared to detect pain when scored from video footage of rabbits before and after orthopedic surgery. In the presence of the observer, the pain scores were underestimated at the time considered to be associated with the greatest pain and overestimated at the times of little or no pain.

Predictors of germline status for hereditary melanoma: 5 years of multi-gene panel testing within the Italian Melanoma Intergroup.

BACKGROUND: The incidence of cutaneous melanoma is increasing in Italy, in parallel with the implementation of gene panels. Therefore, a revision of national genetic assessment criteria for hereditary melanoma may be needed. The aim of this study was to identify predictors of susceptibility variants in the largest prospective cohort of Italian high-risk melanoma cases studied to date. MATERIALS AND METHODS: From 25 Italian centers, we recruited 1044 family members and germline sequenced 940 cutaneous melanoma index cases through a shared gene panel, which included the following genes: CDKN2A, CDK4, BAP1, POT1, ACD, TERF2IP, MITF and ATM. We assessed detection rate according to familial status, region of origin, number of melanomas and presence and type of non-melanoma tumors. RESULTS: The overall detection rate was 9.47% (5.53% analyzing CDKN2A alone), ranging from 5.14% in sporadic multiple melanoma cases (spoMPM) with two cutaneous melanomas to 13.9% in familial cases with at least three affected members. Three or more cutaneous melanomas in spoMPM cases, pancreatic cancer and region of origin predicted germline status [odds ratio (OR) = 3.23, 3.15, 2.43, P < 0.05]. Conversely, age > 60 years was a negative independent predictor (OR = 0.13, P = 0.008), and was the age category with the lowest detection rate, especially for CDKN2A. Detection rate was 19% when cutaneous melanoma and pancreatic cancer clustered together. CONCLUSIONS: Gene panel doubled the detection rate given by CDKN2A alone. National genetic testing criteria may need a revision, especially regarding age cut-off (60) in the absence of strong family history, pancreatic cancer and/or a high number of cutaneous melanomas.

Modified tibial plateau levelling osteotomy to treat lateral patellar luxation and cranial cruciate ligament deficiency in a dog.

The use of modified tibial plateau leveling osteotomy adapted for correction of the lateral patellar dislocation was effective, allowing simultaneous treatment of lateral patellar luxation and cranial cruciate ligament deficiency.

Germline ATM variants predispose to melanoma: a joint analysis across the GenoMEL and MelaNostrum consortia.

PURPOSE: Ataxia-Telangiectasia Mutated (ATM) has been implicated in the risk of several cancers, but establishing a causal relationship is often challenging. Although ATM single-nucleotide polymorphisms have been linked to melanoma, few functional alleles have been identified. Therefore, ATM impact on melanoma predisposition is unclear. METHODS: From 22 American, Australian, and European sites, we collected 2,104 familial, multiple primary (MPM), and sporadic melanoma cases who underwent ATM genotyping via panel, exome, or genome sequencing, and compared the allele frequency (AF) of selected ATM variants classified as loss-of-function (LOF) and variants of uncertain significance (VUS) between this cohort and the gnomAD non-Finnish European (NFE) data set. RESULTS: LOF variants were more represented in our study cohort than in gnomAD NFE, both in all (AF = 0.005 and 0.002, OR = 2.6, 95% CI = 1.56-4.11, p < 0.01), and familial + MPM cases (AF = 0.0054 and 0.002, OR = 2.97, p < 0.01). Similarly, VUS were enriched in all (AF = 0.046 and 0.033, OR = 1.41, 95% CI = 1.6-5.09, p < 0.01) and familial + MPM cases (AF = 0.053 and 0.033, OR = 1.63, p < 0.01). In a case-control comparison of two centers that provided 1,446 controls, LOF and VUS were enriched in familial + MPM cases (p = 0.027, p = 0.018). CONCLUSION: This study, describing the largest multicenter melanoma cohort investigated for ATM germline variants, supports the role of ATM as a melanoma predisposition gene, with LOF variants suggesting a moderate-risk.

Effects of lumbar myelography on subarachnoid pressure and cardiorespiratory parameters in healthy sheep / Efeitos da mielografia lombar na pressão subaracnoidea e nos parâmetros cardiorrespiratórios em ovelhas saudáveis

The purpose of this study was to evaluate the effects of lumbar myelography on subarachnoid pressure, cardiorespiratory parameters and pressure-volume index in sheep. Eight sheep were evaluated. The animals were submitted to puncture of the cisterna magna for monitoring of subarachnoid pressure (SaP). Lumbar myelography was performed through applying Iohexol (0.4mL/kg). ToC, PAS, PAD, MAP, InspISO, FeISO, SaP and CPP were recorded immediately after anesthetic stabilization (M0), during lumbar puncture (M1), and two, four and six minutes after contrast application (M2, M3 and M4, respectively). Blood pressure initially increased, then decreased, returning to basal level, similarly observed for InspISo and FeISo. The SaP rose initially thereafter remaining stable. Despite of the effect on subarachnoid pressure, lumbar myelography can be considered safe in sheep.(AU)

O objetivo foi avaliar os efeitos da mielografia lombar na pressão subaracnóidea, parâmetros cardiorrespiratórios e índice pressão-volume em ovinos. Oito ovelhas foram avaliadas. Os animais foram submetidos à punção da cisterna magna para monitoramento da pressão subaracnoidea (SaP). A mielografia lombar foi realizada com a aplicação de Iohexol (0,4mL/kg). Os momentos estudados foram: imediatamente após a estabilização anestésica (M0), durante a punção lombar (M1), dois, quatro e seis minutos após a aplicação do contraste (M2, M3 e M4, respectivamente), para avaliar as seguintes variáveis: ToC, PAS, PAD, MAP, InspISO, FeISO, Sap e CPP. A pressão arterial aumentou inicialmente, depois diminuiu, retornando aos valores iniciais, padrão também observado para o InspISo e o FeISo. O SaP subiu inicialmente, posteriormente, permanecendo estável. A mielografia lombar, apesar de influenciar a pressão subaracnoidea, pode ser considerada segura em ovinos.(AU)

Effects of lumbar myelography on subarachnoid pressure and cardiorespiratory parameters in healthy sheep

ABSTRACT: The purpose of this study was to evaluate the effects of lumbar myelography on subarachnoid pressure, cardiorespiratory parameters and pressure-volume index in sheep. Eight sheep were evaluated. The animals were submitted to puncture of the cisterna magna for monitoring of subarachnoid pressure (SaP). Lumbar myelography was performed through applying Iohexol (0.4mL/kg). ToC, PAS, PAD, MAP, InspISO, FeISO, SaP and CPP were recorded immediately after anesthetic stabilization (M0), during lumbar puncture (M1), and two, four and six minutes after contrast application (M2, M3 and M4, respectively). Blood pressure initially increased, then decreased, returning to basal level, similarly observed for InspISo and FeISo. The SaP rose initially thereafter remaining stable. Despite of the effect on subarachnoid pressure, lumbar myelography can be considered safe in sheep.

RESUMO: O objetivo foi avaliar os efeitos da mielografia lombar na pressão subaracnóidea, parâmetros cardiorrespiratórios e índice pressão-volume em ovinos. Oito ovelhas foram avaliadas. Os animais foram submetidos à punção da cisterna magna para monitoramento da pressão subaracnoidea (SaP). A mielografia lombar foi realizada com a aplicação de Iohexol (0,4mL/kg). Os momentos estudados foram: imediatamente após a estabilização anestésica (M0), durante a punção lombar (M1), dois, quatro e seis minutos após a aplicação do contraste (M2, M3 e M4, respectivamente), para avaliar as seguintes variáveis: ToC, PAS, PAD, MAP, InspISO, FeISO, Sap e CPP. A pressão arterial aumentou inicialmente, depois diminuiu, retornando aos valores iniciais, padrão também observado para o InspISo e o FeISo. O SaP subiu inicialmente, posteriormente, permanecendo estável. A mielografia lombar, apesar de influenciar a pressão subaracnoidea, pode ser considerada segura em ovinos.

Proteolytic activity of Triatoma infestans saliva associated with PAR-2 activation and vasodilation

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect and the main vector of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). In the present study, the authors investigated whether a serine protease activity from the saliva of T. infestans has a role in vasomotor modulation, and in the insect-blood feeding by cleaving and activating protease-activated receptors (PARs). Methods T. infestans saliva was chromatographed as previously reported for purification of triapsin, a serine protease. The cleavage activity of triapsin on PAR peptides was investigated based on FRET technology. Mass spectrometry was used to analyze the sites of PAR-2 peptide cleaved by triapsin. NO measurements were performed using the DAN assay (2,3-diaminonapthalene). The vasorelaxant activity of triapsin was measured in vessels with or without functional endothelium pre-contracted with phenylephrine (3 µM). Intravital microscopy was used to assess the effect of triapsin on mouse skin microcirculation. Results Triapsin was able to induce hydrolysis of PAR peptides and showed a higher preference for cleavage of the PAR-2 peptide. Analysis by mass spectrometry confirmed a single cleavage site, which corresponds to the activation site of the PAR-2 receptor. Triapsin induced dose-dependent NO release in cultured human umbilical vein endothelial cells (HUVECs), reaching a maximum effect at 17.58 nM. Triapsin purified by gel-filtration chromatography (10-16 to 10-9 M) was applied cumulatively to mouse mesenteric artery rings and showed a potent endothelium-dependent vasodilator effect (EC30 = 10-12 M). Nitric oxide seems to be partially responsible for this vasodilator effect because L-NAME (L-NG-nitroarginine methyl ester 300 µM), a nitric oxide synthetase inhibitor, did not abrogate the vasodilation activated by triapsin. Anti-PAR-2 antibody completely inhibited vasodilation observed in the presence of triapsin activity. Triapsin activity also induced an increase in the mouse ear venular diameter. Conclusion Data from this study suggest a plausible association between triapsin activity mediated PAR-2 activation and vasodilation caused by T. infestans saliva.(AU)

Lyophilized and gamma-sterilized allogeneic bone implant used as a spacer for advancement of a modified tibial tuberosity in the treatment of cranial cruciate ligament disease in dogs.

The objective of this study was to evaluate the use of a lyophilized and gamma-sterilized allogeneic freeze-dried bone wedge as a spacer for advancement of a modified tibial tuberosity (mTTA) in 16 knees that were clinically diagnosed with cranial cruciate ligament disease. Patients underwent radiography before the surgical procedure as well as immediately after surgery and at 30, 60, 90 and 120 days post-surgery, and their locomotion was evaluated at the same time points except for the immediate postoperative period. The surgical wounds were evaluated for signs of infection and rejection of the bone implant. Locomotion was graded on a scale of 0-5, with 0 indicating no limping and 5 indicating limb functional impotence. The "tibial-tibial bone-tibial implant" interfaces were evaluated radiographically, and each interface was assigned scores of 0-3, with 0 indicating no contact between the implant and adjacent bone and 3 indicating a bone bridge throughout the interface. The patients showed good clinical and radiographic recovery. The lyophilized bone spacer allowed for easy storage and transport and rapid and satisfactory execution of mTTA while showing resistance to drilling and fixation with screws in 87.5% of cases and a mean surgical time of 45.9 minutes. No immunogenic reactions were observed in 93.7% of the cases. One patient presented infection of the surgical focus, which showed remission after antimicrobial therapy. All patients showed functional recovery of the operated limb, with the number of clinically healthy patients being higher than those with claudication at 120 days (p ≤ 0.05). In all patients, it was possible to verify the incorporation of the bone implant into the tibia. Bone union occurred progressively, and the degrees of bone union observed on radiographs at postoperative days 60, 90, and 120 were significantly greater (p < 0.05) than those observed in the immediate postoperative period and at 30 days.

Minimally invasive hysterectomy in Coatis ( Nasua nasua ) / Histerectomia minimamente invasiva em Quatis (Nasua nasua)

Some wildlife species, such as coatis, have a high degree of adaptability to adverse conditions, such as fragmented urban forests, increasingly common on the world stage. The increase in the number of these mesopredators causes drastic changes in the communities of smaller predators, interferes with reproductive success of trees, as well as becoming a form of exchange between domestic and wild areas, favoring the transmission of zoonosis and increasing the occurrence of attacks to animals or people. This report describes the use of minimally invasive hysterectomy in two individuals of the species Nasua nasua, which can be accomplished through the use of hook technique, commonly used to castrate dogs and cats. The small incision and healing speed of incised tissues are fundamental in wild life management since the postoperative care is limited by the behavior of these animals. This technique proved to be effective and can greatly reduce the morbidity of this procedure in coatis.(AU)

Algumas espécies selvagens, como os quatis, possuem alto grau de adaptabilidade a condições adversas, como as de florestas urbanas fragmentadas, cada vez mais comuns no cenário mundial. O aumento do número destes mesopredadores causa alterações drásticas nas comunidades de pequenos predadores, interfere no sucesso reprodutivo de árvores, além de se tornar uma forma de intercâmbio entre áreas domésticas e selvagens, facilitando a veiculação de zoonoses e aumentando a ocorrência de ataques a animais ou pessoas. O presente relato descreve a utilização da histerectomia minimamente invasiva em dois indivíduos da espécie Nasua nasua, que pode ser realizada por meio da utilização da técnica do gancho, comumente utilizada para a castração de cães e gatos. A incisão reduzida e a rapidez da cicatrização dos tecidos incisados são fundamentais no manejo de espécies selvagens, visto que os cuidados pós-operatórios são limitados pelo comportamento desses animais. Esta técnica mostrou-se eficaz e pode reduzir sobremaneira a morbidade desse procedimento em quatis.(AU)

Origem e distribuição das principais artérias do membro torácico de Cuniculus paca (Linnaeus, 1766) / Origin and distribution of the main arteries of the thoracic limb of Cuniculus paca (Linnaeus, 1766)

Objetivou-se descrever as artérias do membro torácico da paca (Cuniculos paca Linanaeus, 1766), mediante a dissecação da região. Para tanto, foram utilizadas 10 pacas adultas, machos ou fêmeas, pesando entre cinco e 10 kg do plantel de pacas do setor de Animais Silvestres da FCAV, Unesp, Jaboticabal-SP. Nos animais, injetou-se látex pela artéria carótida comum esquerda para preencher e corar todo o sistema arterial, seguido pela fixação em formaldeído a 10% e conservação em solução salina a 30% para dissecação anatômica das principais artérias do arco aórtico, braço e antebraço, identificando-se a origem e distribuição destes vasos. Os resultados foram foto documentados e discutidos com base na literatura sobre os animais domésticos, e roedores selvagens. De forma geral, as artérias do membro torácico da paca, assemelham-se com as dos carnívoros domésticos, do rato e da cobaia.(AU)

The aim of this study is to describe the arteries of the thoracic limb of paca (Cuniculos paca Linanaeus, 1766) by dissecting the region. We used 10 adult paca, males and females, weighting between five to 10 kg obtained from the breeding group at the Wild Animals Sector of FCAV, Unesp, Jaboticabal-SP. The animals were injected with latex by the left common carotid artery to fill and stain all the arterial system, followed by the fixation in 10% formaldehyde and preservation in 30% saline solution for anatomical dissection of the main arteries of the aortic arch, arm, forearm, identifying the origin and distribution of these vessels. The results were photodocumented and discussed based on literature about domestic animals and wild rodent. In general, the arteries of the thoracic limb of paca are similar to the domestic carnivores, rat and guinea pig.(AU)

Miologia do membro pélvico da paca (Cuniculus paca Linnaeus, 1766) / The myology of the pelvic limb of paca (Cuniculus paca Linnaeus, 1766)

Objetivam-se descrever os músculos do membro pélvico da paca (Cuniculus paca Linnaeus, 1766), mediante dissecção anatômica dessa região. Foram utilizadas dez Cuniculus paca adultas, machos e fêmeas, pesando entre cinco e 10 kg do plantel de pacas do setor de Animais Silvestres da FCAV, Unesp, Jaboticabal/SP. Os animais foram fixados em formaldeído 10% e conservados em solução salina a 30% para dissecação anatômica da musculatura da pelve, coxa e perna, identificando-se a origem e inserção de cada músculo. Os resultados foram fotodocumentados e discutidos com base na literatura sobre animais domésticos, ratos e cobaias. Salvo algumas variâncias na origem e inserção de cada músculo e na fusão dos ventres de alguns grupos musculares, de forma geral, a musculatura da paca assemelha-se a dos animais domésticos e a de outros roedores.(AU)

This study aimed to describe the muscles of the pelvic limb of paca (Cuniculus paca Linnaeus, 1766) through anatomical dissection of this region. We used ten adult pacas, males and females, weighing 5 to 10 kg from the group of pacas at the sector of wild animals of FCAV, Unesp-Jaboticabal, SP. The animals were fixed with 10% formaldehyde and preserved in 30% saline solution for anatomical dissection of the pelvis, thigh and leg musculature, with identification of the origin and the insertion of each muscle. Photos were taken and the results were discussed based on the domestic animal, rat and guinea pig literature. In general, the paca musculature resembles the one of domestic animals and other rodents; but some variance in origin and insertion of each muscle and in the fusion of some muscular groups of the bellies was observed.(AU)

Aspecto radiológico e macroscópico de matriz óssea mineralizada heteróloga fragmentada e polimetilmetacrilato autoclavados em falha óssea de tíbia de coelho / Radiological and macroscopic aspects of autoclaved heterologous fragmented mineralized bone matrix and polymethylmethacrylate in bone defect in tibia of rabbits

Foi realizada falha segmentar com 6mm de diâmetro na região metafisária medial de tíbias de 12 coelhos, onde foi implantado uma associação de micro e macrofragmentos de matriz óssea cortical heteróloga fragmentada conservada em glicerina (98%) e polimetilmetacrilato autoclavados, para a sua reconstrução, e avaliados radiológica e macroscopicamente aos 30, 60, 90 e 120 dias. Houve adesão, em relação ao tempo, dos micro e macrocompósitos ao leito receptor, em 100% dos casos, mostrando ser biologicamente biocompatível, pois promoveram a reparação de falhas ósseas, sem sinais de infecção, migração e/ou rejeição, podendo, dessa forma, ser mais uma opção como substituto para preencher grandes defeitos ósseos.

A segmental defect of 6mm diameter was performed in the medial metaphyseal region of the tibia of 12 rabbits. For the bone defect reconstruction there was implanted a combination of micro and macro fragments of heterologous fragmented cortical bone matrix preserved in glycerin (98%) and polymethylmethacrylate, both autoclaved. Radiological and macroscopic evaluation was performed at 30, 60, 90 and 120 days. Adhesion in relation to time of the micro and macro composites to the recipient bed was observed in 100% of the cases. This showed that this implant is biologically biocompatible, since it promoted bone defects repair, with no signs of infection, migration and/or rejection. In this way, this implant can be classified as one more option of substitute to fill large bone defects.

Assessment of postoperative pain after unilateral mastectomy using two different surgical techniques in dogs.

BACKGROUND: There are few studies reporting pain and postoperative analgesia associated with mastectomy in dogs. The aim of this study was to evaluate postoperative pain after unilateral mastectomy using two different surgical techniques in the dog. FINDINGS: Twenty female dogs were assigned (n=10/group) to undergo unilateral mastectomy using either the combination of sharp and blunt dissection (SBD) or the modified SBD (mSBD) technique, in which the mammary chain is separated from the abdominal wall entirely by blunt (hand and finger) dissection except for a small area cranial to the first gland, in a prospective, randomized, clinical trial. All dogs were premedicated with intramuscular acepromazine (0.05 mg/kg) and morphine (0.3 mg/kg). Anesthesia was induced with intravenous ketamine (5 mg/kg) and diazepam (0.25 mg/kg), and maintained with isoflurane. Subcutaneous meloxicam (0.2 mg/kg) was administered before surgery. Postoperative pain was evaluated according to the University of Melbourne pain scale (UMPS) by an observer who was blinded to the surgical technique.. Rescue analgesia was provided by the administration of intramuscular morphine (0.5 mg/kg) if pain scores were >14 according to the UMPS. Data were analyzed using t-tests and ANOVA (P>0.05). There were no significant differences between the groups for age, weight, extubation time, and duration of surgery and anesthesia (P>0.05). There were no significant differences for postoperative pain scores between groups. Rescue analgesia was required in one dog in each group. CONCLUSIONS: The two surgical techniques produced similar surgical times, incidence of perioperative complications and postoperative pain. Multimodal analgesia is recommended for treatment of postoperative pain in dogs undergoing unilateral mastectomy.

Prevalence of the E318K MITF germline mutation in Italian melanoma patients: associations with histological subtypes and family cancer history.

A French and an Australian study have recently identified a rare germline functional variant in the microphthalmia-associated transcription factor (MITF) (E318K) that predisposes to familial and sporadic melanoma and to renal cell carcinoma (RCC), showing a new link between two tumour types with different risk factors and between deregulated sumoylation and cancer. The aim of this study was to test the prevalence of the MITF E318K mutation in 667 Italian melanoma patients. We observed significant associations between histological subtypes and family cancer history. Carriers exhibited a nearly threefold higher risk of developing melanoma compared with controls. Carriers were also more likely to have developed multiple primary melanomas (6.40-fold), compared with wt patients. Carriers with a personal and/or family history of pancreatic cancer and kidney cancer had a nearly 31- and eightfold higher risk of developing melanoma compared with wt patients. Our findings further support MITF as a medium-penetrance melanoma susceptibility gene, highlight a potential association with histological subtypes and suggest that MITF may predispose to pancreatic cancer.

Helicobacter pylori colonization ameliorates glucose homeostasis in mice through a PPAR γ-dependent mechanism.

BACKGROUND: There is an inverse secular trend between the incidence of obesity and gastric colonization with Helicobacter pylori, a bacterium that can affect the secretion of gastric hormones that relate to energy homeostasis. H. pylori strains that carry the cag pathogenicity island (PAI) interact more intimately with gastric epithelial cells and trigger more extensive host responses than cag(-) strains. We hypothesized that gastric colonization with H. pylori strains differing in cag PAI status exert distinct effects on metabolic and inflammatory phenotypes. METHODOLOGY/PRINCIPAL FINDINGS: To test this hypothesis, we examined metabolic and inflammatory markers in db/db mice and mice with diet-induced obesity experimentally infected with isogenic forms of H. pylori strain 26695: the cag PAI wild-type and its cag PAI mutant strain 99-305. H. pylori colonization decreased fasting blood glucose levels, increased levels of leptin, improved glucose tolerance, and suppressed weight gain. A response found in both wild-type and mutant H. pylori strain-infected mice included decreased white adipose tissue macrophages (ATM) and increased adipose tissue regulatory T cells (Treg) cells. Gene expression analyses demonstrated upregulation of gastric PPAR γ-responsive genes (i.e., CD36 and FABP4) in H. pylori-infected mice. The loss of PPAR γ in immune and epithelial cells in mice impaired the ability of H. pylori to favorably modulate glucose homeostasis and ATM infiltration during high fat feeding. CONCLUSIONS/SIGNIFICANCE: Gastric infection with some commensal strains of H. pylori ameliorates glucose homeostasis in mice through a PPAR γ-dependent mechanism and modulates macrophage and Treg cell infiltration into the abdominal white adipose tissue.

Contribution of germline mutations in the BRCA and PALB2 genes to pancreatic cancer in Italy.

Pancreatic adenocarcinoma (PC) is the third most common cancer associated with BRCA mutations. Most notice has been given to BRCA2, while the association between BRCA1 and PC is less widely reported. Recently, PALB2 has been implicated in both PC and breast cancer (BC) susceptibility. We selected 29 Italian PC patients from a case-control study of PC according to their personal and family history of both PC and breast/ovarian cancer (BC/OC) and tested them for presence of germline mutations in BRCA1, BRCA2 and PALB2. We identified no germline mutations or deletions in PALB2, but detected 7 BRCA mutations (4 in BRCA1 and 3 in BRCA2). These findings suggest that PALB2 does not play a major role in PC susceptibility in our population. As we found an almost equal frequency of germline mutations in BRCA1 and BRCA2, germline alterations in either of these genes may explain a subset of Italian families presenting both PC and BC/OC. Moreover, as we began the observation of these families from probands who are affected by PC, we provide here a direct assessment of the role of PALB2 and BRCA mutations in PC susceptibility.