Caracterização demográfica das populações de cães e gatos supervisionados do município de São Paulo

A caracterização das populações canina e felina domiciliadas do município de São Paulo (SP) foi realizada utilizando-se amostragem complexa com seleção aleatória em dois estágios. Em cada distrito administrativo, foram visitados seis setores censitários e 20 domicílios em cada setor sorteado, de setembro de 2006 a setembro de 2009, totalizando 11.272 entrevistas. A razão homem:cão foi de 4,34, e a razão homem:gato de 19,33. A população canina foi estimada em 2.507.401, e a felina em 562.965. A população canina era composta por 52,7% de machos, e a felina por 45,1%. A proporção de fêmeas esterilizadas, 23,4% das cadelas e 46,1% das gatas, é maior que a de machos, 11,4% dos cães e 31,5% dos gatos. A idade média dos cães era de 4,99 anos e a dos gatos de 3,53 anos. A proporção de cães com restrição de acesso à rua, 64,4%, foi maior que a de gatos, 42,5%. A média de animais/domicílio foi estimada em 1,60 para cães e 1,69 para gatos. A guarda destes animais está associada a fatores culturais, assim a caracterização da população de animais é base da estruturação adequada de programas de controle populacional e de zoonoses.

Reposição volêmica com soluções salinas em pancreatite e perfil hepático de proteínas apoptóticas e de choque térmico

OBJETIVO: A falência hepática é uma consequência da inflamação sistêmica após pancreatite aguda. Avaliou-se o efeito da reposição volêmica com soluções salinas fisiológicas ou hipertônica na produção hepática de citocinas e na expressão de proteínas ativadas por choque térmico e proteínas ligadas à apoptose durante a pancreatite aguda. MÉTODOS: Ratos Wistar foram divididos em quatro grupos: C - animais controles não submetidos à lesão e nem ao tratamento; NT - animais submetidos à indução de pancreatite aguda e não tratados; SN - animais submetidos à indução de pancreatite aguda e tratados com solução salina normal (NaCl 0,9%); SH - animais submetidos à pancreatite aguda e tratados com solução salina hipertônica (NaCl 7,5%). A pancreatite aguda foi induzida por infusão retrógrada transduodenal de taurocolato de sódio 2,5% no ducto pancreático. Após 4, 12 e 24 horas da indução da pancreatite aguda, analisaram-se, no fígado, TNF-α, IL-1β, IL-6 e IL-10, caspase-2, caspase-7, APAF-1, AIF, HSP60 e HSP90. RESULTADOS: A caspase-2 diminuiu nos grupos SN e SH (p<0,05 versus C) após 12 horas. APAF-1, AIF e HSP90 permaneceram inalterados. Após 4 horas da indução, a capsase-7 aumentou no grupo NT (p<0,01 versus C), embora se mantendo em níveis basais nos grupos reperfundidos. A HSP60 aumentou em todos os grupos após 4 horas (p<0,001 versus C). No entanto, o grupo SH mostrou menor expressão de HSP60 que o grupo SN (p<0,05). A solução salina hipertônica manteve a produção de citocinas em níveis normais. A reperfusão com volume com solução salina normal ou hipertônica, modulou significativamente a expressão de caspase-7. CONCLUSÃO: A reposição volêmica com solução salina normal ou hipertônica foi efetiva em reduzir a caspase-7. Entretanto, somente a solução salina hipertônica foi capaz de regular a produção de citocinas e a expressão de HSP60 em todos os momentos analisados.

The relationship between lymphatic vascular density and vascular endothelial growth factor A (VEGF-A) expression with clinical-pathological features and survival in pancreatic adenocarcinomas

Abstract: Background Pancreatic cancer is a rare tumor with an extremely low survival rate. Its known risk factors include the chronic use of tobacco and excessive alcohol consumption and the presence of chronic inflammatory diseases, such as pancreatitis and type 2 diabetes. Angiogenesis and lymphangiogenesis, which have been the focus of recent research, are considered prognostic factors for cancer development. Knowing the angiogenic and lymphangiogenic profiles of a tumor may provide new insights for designing treatments according to the different properties of the tumor. The aim of this study was to evaluate the density of blood and lymphatic vessels, and the expression of VEGF-A, in pancreatic adenocarcinomas, as well as the relationship between blood and lymphatic vascular density and the prognostically important clinical-pathological features of pancreatic tumors. Methods Paraffin blocks containing tumor samples from 100 patients who were diagnosed with pancreatic cancer between 1990 and 2010 were used to construct a tissue microarray. VEGF expression was assessed in these samples by immunohistochemistry. To assess the lymphatic and vascular properties of the tumors, 63 cases that contained sufficient material were sectioned routinely. The sections were then stained with the D2-40 antibody to identify the lymphatic vessels and with a CD34 antibody to identify the blood vessels. The vessels were counted individually with the Leica Application Suite v4 program. All statistical analyses were performed using SPSS 18.0 (Chicago, IL, USA) software, and p values ≤ 0.05 were considered significant. Results In the Cox regression analysis, advanced age (p=0.03) and a history of type 2 diabetes (p=0.014) or chronic pancreatitis (p=0.02) were shown to be prognostic factors for pancreatic cancer. Blood vessel density (BVD) had no relationship with clinical-pathological features or death. Lymphatic vessel density (LVD) was inversely correlated with death (p=0.002), and by Kaplan-Meyer survival analysis, we found a significant association between low LVD (p=0.021), VEGF expression (p=0.023) and low patient survival. Conclusions Pancreatic carcinogenesis is related to a history of chronic inflammatory processes, such as type 2 diabetes and chronic pancreatitis. In pancreatic cancer development, lymphangiogenesis can be considered an early event that enables the dissemination of metastases. VEGF expression and low LVD can be considered as poor prognostic factors as tumors with this profile are fast growing and highly aggressive. Virtual slides. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/5113892881028514

Canine heartworm disease in Porto Velho: first record, distribution map and occurrence of positive mosquitoes

The aim of this study was to make the first report on canine heartworm disease in the state of Rondônia and confirm its transmission in this state. Blood samples were randomly collected from 727 dogs in the city of Porto Velho. The samples were analyzed to search for microfilariae and circulating antigens, using three different techniques: optical microscopy on thick blood smears stained with Giemsa; immunochromatography; and PCR. Mosquitoes were collected inside and outside the homes of all the cases of positive dogs and were tested using PCR to search for DNA of Dirofilaria immitis. Ninety-three blood samples out of 727 (12.8%) were positive according to the immunoassay technique and none according to the thick smear method. Among the 93 positive dogs, 89 (95.7%) were born in Porto Velho. No difference in the frequency of infection was observed between dogs raised indoors and in the yard. PCR on the mosquitoes resulted in only one positive pool. This result shows that the transmission of canine heartworm disease is occurring in the city of Porto Velho and that there is moderate prevalence among the dogs. The techniques of immunochromatography and PCR were more effective for detecting canine heartworm than thick blood smears. The confirmation of canine heartworm disease transmission in Porto Velho places this disease in the ranking for differential diagnosis of pulmonary nodules in humans in Rondônia.

Il Nilotinib nella terapia di prima linea della leucemia mieloide cronica

Background: Nilotinib is a potent and selective BCR-ABL inhibitor. The phase 3 ENESTnd trial demonstrated superior efficacy nilotinib vs imatinib, with higher and faster molecular responses. After 24 months, the rates of progression to accelerated-blastic phase (ABP) were 0.7% and 1.1% with nilotinib 300mg and 400mg BID, respectively, significantly lower compared to imatinib (4.2%). Nilotinib has been approved for the frontline treatment of Ph+ CML. With imatinib 400mg (IRIS trial), the rate of any event and of progression to ABP were higher during the first 3 years. Consequently, a confirmation of the durability of responses to nilotinib beyond 3 years is extremely important. Aims: To evaluate the response and the outcome of patients treated for 3 years with nilotinib 400mg BID as frontline therapy. Methods: A multicentre phase 2 trial was conducted by the GIMEMA CML WP (ClinicalTrials.gov.NCT00481052). Minimum 36-month follow-up data for all patients will be presented. Definitions: Major Molecular Response (MMR): BCR-ABL/ABL ratio <0,1%IS; Complete Molecular Response (CMR): undetectable transcript levels with ≥10,000 ABL transcripts; failures: according to the revised ELN recommendations; events: failures and treatment discontinuation for any reason. All the analysis has been made according to the intention-to-treat principle. Results: 73 patients enrolled: median age 51 years; 45% low, 41% intermediate and 14% high Sokal risk. The cumulative incidence of CCgR at 12 months was 100%. CCgR at each milestone: 78%, 96%, 96%, 95%, 92% at 3, 6, 12, 18 and 24 months, respectively. The overall estimated probability of MMR was 97%, while the rates of MMR at 3, 6, 12, 18 and 24 months were 52%, 66%, 85%, 81% and 82%, respectively. The overall estimated probability of CMR was 79%, while the rates of CMR at 12 and 24 months were 12% and 27%, respectively. No patient achieving a MMR progressed to AP. Only one patient progressed at 6 months to ABP and subsequently died (high Sokal risk, T315I mutation). Adverse events were mostly grade 1 or 2 and manageable with appropriate dose adaptations. During the first 12 months, the mean daily dose was 600-800mg in 74% of patients. The nilotinib last daily dose was as follows: 800mg in 46 (63%) patients, 600mg in 3 (4%) patients and 400mg in 18 (25%), 6 permanent discontinuations. Detail of discontinuation: 1 patient progressed to ABP; 3 patients had recurrent episodes of amylase and/or lipase increase (no pancreatitis); 1 patient had atrial fibrillation (unrelated to study drug) and 1 patient died after 32 months of mental deterioration and starvation (unrelated to study drug). Two patients are currently on imatinib second-line and 2 on dasatinib third-line. With a median follow-up of 39 months, the estimated probability of overall survival, progression-free survival and failure-free survival was 97%, the estimated probability of event-free survival was 91%. Conclusions: The rate of failures was very low during the first 3 years. Responses remain stable. The high rates of responses achieved during the first 12 months are being translated into optimal outcome for most of patients.

Analisi clinica ed ex-vivo dei punti isometrici durante tecnica tightrope nella ricostruzione del legamento crociato craniale del cane

La rottura del Legamento Crociato Craniale (LCCr) rappresenta una delle patologie ortopediche di maggiore riscontro clinico nella specie canina. In seguito a rottura del LCCr si presenta un continuo slittamento craniale della tibia il quale esita in un processo osteoartrosico. La risoluzione chirurgica rappresenta la migliore soluzione terapeutica. Le tecniche chirurgiche extra-articolari con sfruttamento dei punti isometrici del ginocchio si presentano come delle procedure molto diffuse e utilizzate. Questa tesi propone di validare l’uso di un nuovo sistema di navigazione computerizzato-assistito per la valutazione cinematica durante la ricostruzione del LCCr nel cane, ma soprattutto di studiare e confrontare il comportamento e l’efficacia dopo ricostruzione TightRope (TR) in due diverse coppie di punti isometrici. Abbiamo effettuato due analisi in parallelo. La prima eseguendo interventi chirurgici con tecnica TR su 18 casi clinici e sfruttando il punto isometrico del femore (F2) e due diversi punti isometrici della tibia (T2 o T3). L’analisi prevedeva dei controlli postoperatori a 1, 3 e 6 mesi. Ad ogni controllo veniva effettuata una visita ortopedica, esami radiografici, un questionario di valutazione clinico e di soddisfazione del proprietario. Mentre nella ricerca Ex-Vivo abbiamo eseguito dei test su 14 preparati anatomici con l’utilizzo di un sistema di navigazione computerizzato per la rilevazione dei dati. L’analisi prevedeva la valutazione dell’articolazione in diversi stadi: LCCr intatto; LCCr rotto; dopo ricostruzione con TR in F2-T2 e tensionato a 22N, 44N e 99N; dopo ricostruzione con TR in F2-T3 e tensionato a 22N, 44N e 99N. Ad ogni stadio si eseguivano cinque test di valutazione, tra cui: Test del Cassetto, Test di compressione tibiale (TCT), Rotazione Interna/Esterna, Flesso/Estensione e Varo/Valgo. Lo scopo di tale studio è quello di confrontare tra loro i punti isometrici del ginocchio e di analizzare l’efficacia della tecnica TR nelle due differenti condizioni di isometria (F2-T2 e F2-T3).

Presenza e diffusione della Leishmaniosi e dei suoi vettori in provincia di Rieti

Nel presente studio ci si è proposti di valutare la presenza e diffusione della leishmaniosi in una provincia dell’Italia centrale (Rieti) che, per caratteristiche ambientali (prevalentemente montuosa, clima freddo-secco) poco sembra prestarsi al ciclo della malattia. A questo scopo sono stati calcolati: i) sieroprevalenza grezza nella popolazione canina (2006-2013) e prevalenza media annuale ii) casi di leishmaniosi viscerale (LV) e cutanea (LC) (2000-2013). Catture di flebotomi sono state effettuate per due stagioni consecutive (2011-2012) per ogni sito sono stati registrati i dati climatici (temperatura, umidità etc.) ed altitudine. I flebotomi sono stati sottoposti a ricerca di Leishmania mediante PCR. La sieroprevalenza grezza per leishmania varia da 0 a 76,9% e la prevalenza media annuale non presenta un trend lineare. Sono stati registrati 6 casi di LV tutti in pazienti italiani tutti residenti in provincia di Rieti. I flebotomi sono stati rilevati in 5 dei 6 siti monitorati fino agli 800 m s.l.m., seppur con basse densità. Sono state identificate le seguenti specie: P. perniciosus (6,4 %), P. perfiliewi (1,8%), P. mascittii (0,1%) e S. minuta (91,7 %). E’ stata rilevata una correlazione statisticamente significativa (r=0,69, p<0,001) tra numero di flebotomi e temperatura giornaliera (Tmed°C) ed una correlazione negativa significativa (r= -0,51, p<0,05) con l’umidità relativa (Umed%). La ricerca di leishmania ha dato esito negativo in tutti i flebotomi analizzati. Questi rilievi suggeriscono l’endemia della leishmaniosi nella provincia di Rieti.

Hyperglycaemia but not hyperlipidaemia decreases serum amylase and increases neutrophils in the exocrine pancreas of cats

The goal of the study was to determine whether hyperglycaemia or hyperlipidaemia causes pancreatitis in cats and to assess the effect of excess serum glucose and lipids on amylase and lipase activity. Ten-day hyperglycaemic and hyperlipidaemic clamps were carried out in five and six healthy cats, respectively. Ten healthy cats received saline and served as controls. The activity of amylase was below the normal range in 4 of 5 hyperglycaemic cats by day 10. The activity of lipase did not vary in any of the cats. Samples of exocrine pancreas were normal on histological examination, but the number of tissue neutrophils was increased in hyperglycaemic cats (P<0.05). In a retrospective study 14 of 40 (35%) cats with naturally occurring diabetes mellitus had amylase activities below the reference range at the time of admission. Amylase activities normalised within 1 week of insulin therapy and subsequent glycaemic control. Lipase activity was increased in 26 of 40 (65%) diabetic cats and remained elevated despite glycaemic control. In conclusion, hyperglycaemia, but not hyperlipidaemia, increases pancreatic neutrophils in cats. However, because the histological morphology of the exocrine pancreas was normal, hyperglycaemia may play only a minor role in the pathogenesis of pancreatitis. Low amylase activities in diabetic cats may reflect an imbalance in glucose metabolism rather than pancreatitis.

How do we apply video-assisted retroperitoneal necrosectomy with minimal access?

The conservative treatment of acute necrotizing pancreatitis has greatly improved due to broad antibiotic treatment and improved organ support in intensive care units. Nevertheless, infected necrosis or persistent multi-organ dysfunction are predictors of poor outcome. In these patients, there is still a need to perform necrosectomy. Open surgery results in extensive operative trauma and is associated with high morbidity and mortality. Therefore, several minimally invasive techniques have been developed recently. Retroperitoneal necrosectomy has been shown to be safe and to reduce morbidity and mortality compared to the open procedure.

Alcohol dehydrogenase 1C*1 allele is a genetic marker for alcohol-associated cancer in heavy drinkers

Chronic alcohol consumption is associated with an increased risk for upper aerodigestive tract cancer and hepatocellular carcinoma. Increased acetaldehyde production via alcohol dehydrogenase (ADH) has been implicated in the pathogenesis. The allele ADH1C*1 of ADH1C encodes for an enzyme with a high capacity to generate acetaldehyde. So far, the association between the ADH1C*1 allele and alcohol-related cancers among heavy drinkers is controversial. ADH1C genotypes were determined by polymerase chain reaction and restriction fragment length polymorphism in a total of 818 patients with alcohol-associated esophageal (n=123), head and neck (n=84) and hepatocellular cancer (n=86) as well as in patients with alcoholic pancreatitis (n=117), alcoholic liver cirrhosis (n=217), combined liver cirrhosis and pancreatitis (n=17) and in alcoholics without gastrointestinal organ damage (n=174). The ADH1C*1 allele and genotype ADH1C*1/1 were significantly more frequent in patients with alcohol-related cancers than that in individuals with nonmalignant alcohol-related organ damage. Using multivariate analysis, ADH1C*1 allele frequency and rate of homozygosity were significantly associated with an increased risk for alcohol-related cancers (p<0.001 in all instances). The odds ratio for genotype ADH1C*1/1 regarding the development of esophageal, hepatocellular and head and neck cancer were 2.93 (CI, 1.84-4.67), 3.56 (CI, 1.33-9.53) and 2.2 (CI, 1.11-4.36), respectively. The data identify genotype ADH1C*1/1 as an independent risk factor for the development of alcohol-associated tumors among heavy drinkers, indicating a genetic predisposition of individuals carrying this genotype.

Selective in vitro targeting of GRP and NMB receptors in human tumours with the new bombesin tracer 177Lu-AMBA

PURPOSE: To investigate the in vitro binding properties of a novel radiolabelled bombesin analogue, (177)Lu-AMBA, in human neoplastic and non-neoplastic tissues selected for their expression of the bombesin receptor subtypes GRP-R, NMB-R and BRS-3. METHODS: In vitro receptor autoradiography was performed in cancers expressing the various bombesin receptor subtypes. The novel radioligand (177)Lu-AMBA was used and compared with established bombesin radioligands such as (125)I-Tyr(4)-bombesin and (125)I-[D: -Tyr(6),beta-Ala(11),Phe(13),Nle(14)]-bombesin(6-14). In vitro incidence of detection of each of the three bombesin receptor subtypes was evaluated in each tumour. RESULTS: (177)Lu-AMBA identified all GRP-R-expressing tumours, such as prostatic, mammary and renal cell carcinomas as well as gastrointestinal stromal tumours. (177)Lu-AMBA also identified all NMB-expressing tumours, but did not detect BRS-3-expressing tumours or BRS-3-expressing pancreatic islets. GRP-R-expressing peritumoural vessels were heavily labelled with (177)Lu-AMBA. In contrast to the strongly GRP-R-positive mouse pancreas, the human pancreas was not labelled with (177)Lu-AMBA unless chronic pancreatitis was diagnosed. In general, the sensitivity was slightly better with (177)Lu-AMBA than with the conventional bombesin radioligands. CONCLUSION: The present in vitro study suggests that (177)Lu-AMBA may be a very useful in vivo targeting agent for GRP-R-expressing tumours, NMB-R-expressing tumours and GRP-R-expressing neoangiogenic vessels.

A novel secretin receptor splice variant potentially useful for early diagnosis of pancreatic carcinoma

BACKGROUND ; AIMS: Pancreatic and bile duct carcinomas represent highly aggressive malignancies that evolve from secretin receptor-rich ductular cells. With premessenger RNA splicing abnormalities common in cancer, we evaluated whether an abnormal secretin receptor spliceoform were present, characterized it, and developed a serum assay for it. METHODS: Cancer cell lines and healthy and neoplastic tissue were studied by nested reverse-transcription polymerase chain reaction and sequencing. A promising spliceoform was isolated and characterized, and monoclonal antibodies were raised to 2 distinct regions. A dual antibody enzyme-linked immunosorbent assay was developed and applied to blinded serum samples from 26 patients with pancreatic carcinoma, 10 patients with chronic pancreatitis, and 14 controls. RESULTS: Each of 9 pancreatic cancer specimens and no normal tissue expressed a secretin receptor variant with exons 3 and 4 deleted. This encoded a 111-residue peptide with its first 43 residues identical to wild-type receptor, but, subsequent to a shift in coding frame and early truncation, the next 68 residues were unique in the transcriptome/proteome. This nonfunctional soluble protein did not bind or signal in response to secretin and was secreted from transfected MiaPaCa-2 cells. Elevated serum levels of this variant were present in 69% of pancreatic cancer patients, 60% of chronic pancreatitis patients, and 1 of 14 controls. CONCLUSIONS: We identified a novel abnormal spliceoform of the secretin receptor in pancreatic and bile duct cancers and developed a dual antibody sandwich enzyme-linked immunosorbent assay to measure it in the circulation. Initial application of this assay in patients with pancreatic cancer and chronic pancreatitis was promising, but additional validation will be required to evaluate its clinical utility.

Mumps

Mumps is a common childhood infection caused by the mumps virus. The hallmark of infection is swelling of the parotid gland. Aseptic meningitis and encephalitis are common complications of mumps together with orchitis and oophoritis, which can arise in adult men and women, respectively; other complications include deafness and pancreatitis. Clinical diagnosis can be based on the classic parotid swelling; however, this feature is not present in all cases of mumps and can also occur in various other disorders. Laboratory diagnosis is based on isolation of virus, detection of viral nucleic acid, or serological confirmation (generally presence of IgM mumps antibodies). Mumps is vaccine-preventable, and one dose of mumps vaccine is about 80% effective against the disease. Routine vaccination has proven highly effective in reducing the incidence of mumps, and is presently used by most developed countries; however, there have been outbreaks of disease in vaccinated populations. In 2005, a large epidemic peaked in the UK, and in 2006 the American midwest had several outbreaks. In both countries, the largest proportion of cases was in young adults. In the UK, susceptible cohorts too old to have been vaccinated and too young to have been exposed to natural infections were the primary cause of the mumps epidemic. In the USA, effectiveness and uptake in combination appear not to have been sufficient to obtain herd immunity for mumps in populations such as college students.

The role of common single-nucleotide polymorphisms on exon 9 and exon 12 skipping in nonmutated CFTR alleles

Classic cystic fibrosis (CF) is caused by two loss-of-function mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, whereas patients with nonclassic CF have at least one copy of a mutant gene that retains partial function of the CFTR protein. In addition, there are several other phenotypes associated with CFTR gene mutations, such as idiopathic chronic pancreatitis. In CFTR-associated disorders and in nonclassic CF, often only one CFTR mutation or no CFTR mutations can be detected. In this study, we screened 23 patients with CFTR-associated disorders for CFTR mutations by complete gene testing and quantitative transcript analysis. Mutations were found in 10 patients. In cells from respiratory epithelium, we detected aberrant splicing of CFTR mRNA in all investigated individuals. We observed a highly significant association between the presence of coding single-nucleotide polymorphisms (coding SNPs, or cSNPs) and increased skipping of exon 9 and 12. This association was found both in patients and in normal individuals carrying the same cSNPs. The cSNPs c.1540A>G, c.2694T>G, and c.4521G>A may have affected pre-mRNA splicing by changing regulatory sequence motifs of exonic splice enhancers, leading to lower amounts of normal transcripts. The analysis of CFTR exons indicated that less frequent and weak exonic splicing enhancer (ESE) motifs make exon 12 vulnerable to skipping. The number of splice variants in individuals with cSNPs was similar to previously reported values for the T5 allele, suggesting that cSNPs may enhance susceptibility to CFTR related diseases. In addition, cSNPs may be responsible for variation in the phenotypic expression of CFTR mutations. Quantitative approaches rather than conventional genomic analysis are required to interpret the role of cSNPs.

[Odynophagia of a young mother]

We report the case of a 34-year old mother presenting in the emergency unit because of fever and odynophagia. Herpetic esophagitis with Herpes simplex virus primoinfection in a immunocompetent patient was diagnosed. The virus was transmitted by her own child which came to be hospitalised because of herpetic stomatitis. In the blood chemistry pancreas enzymes were elevated which we interpreted as herpetic pancreatitis because of coincidence with herpetic esophagitis.