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Multiple myeloma (MM) has a highly heterogeneous genetic background, which complicates its molecular tracking over time. Nevertheless, each MM patient’s malignant plasma cells (PCs) share unique V(D)J rearranged sequences at immunoglobulin loci, which represent ideal disease biomarkers. Because the tumor-specific V(D)J sequence is highly expressed in bulk RNA in MM patients, we wondered whether it can be identified by single-cell RNA sequencing (scRNA-seq). To this end we analyzed CD138+ cells purified from bone marrow aspirates of 19 samples with PC dyscrasias by both a standard method based on bulk DNA and by an implementation of the standard 10x Genomics protocol to detect expressed V(D)J sequences. A dominant clonotype was easily identified in each sample, accounting on average for 83.65% of V(D)J-rearranged cells. Compared with standard methods, scRNA-seq analysis proved highly concordant and even more effective in identifying clonal productive rearrangements, by-passing limitations related to the misannealing of consensus primers in hypermutated regions. We next validated its accuracy to track 5 clonal cells with absolute sensitivity in a virtual sample containing 3180 polyclonal cells. This shows that single-cell V(D)J analysis may be used to find rare clonal cells, laying the foundations for functional single-cell dissection of minimal residual disease.

Monoclonal gammopathy of undetermined significance (MGUS) and clonal hematopoiesis (CH) are 2 preclinical clonal expansions of hematopoietic cells whose prevalence rises with age, reaching almost 10% in people of aged 70 years and older. The increased risk of myeloid malignancies in patients with myeloma is well defined, and the study of the association between CH and MGUS could help explain this phenomenon. Here, we analyzed a fully clinically annotated dataset of 777 older subjects (median age, 91 years) previously screened for prevalence of CH. The prevalence of MGUS and CH was 9.6% and 17.3%, respectively. We detected CH in 9.7% of the patients with MGUS and MGUS in 5.5% of the patients with CH. We did not find a significant correlation between the presence of MGUS and CH. Furthermore, the 2 conditions showed a differential association with clinical and laboratory covariates, suggesting that MGUS and CH may represent age-associated unrelated clonal drifts of hematopoietic cells. Confirmatory studies are needed to assess the relevance of CH in plasma cell disorders. This trial was registered at www.clinicaltrials.gov as #NCT03907553.

Clonal, pre-clinical expansions of hematopoietic cells are increasingly recognized. Clonal hematopoiesis of indeterminate potential (CHIP) has been recently described. It is characterized by the presence of mutations usually involved in myeloid neoplasia in people without any sign of hematological malignancy. Well before CHIP, another pre-clinical clonal expansion has been described for plasma cells (PC) based on the presence of a serum monoclonal protein, namely monoclonal gammopathy of undetermined significance (MGUS). Despite both being frequent, the association between these two bone marrow (BM) clonal entities has not been determined yet. Since the incidence of MGUS and CHIP rises with age reaching >10% of people >70y, we analyzed a unique oldest-old population dataset to define a possible association between these two different BM clonal disorders. We analyzed a cohort of 777 patients with a median age of 91 years (range, 81 - 104), significantly higher than previous studies on MGUS. 579 (74.5%) were females and 198 (25.5%) males. CHIP was assessed in all but 45 through sequencing of a myeloid specific gene panel. Serum protein electrophoresis was available in all of them to assess the prevalence of MGUS. The prevalence of CHIP and MGUS were 17.5% (128/732) and 9.5% (74/777), respectively. Importantly, CHIP and MGUS did not associate in our cohort but rather showed a non-significant trend towards anti-correlation (Fisher's Exact test, p-val = 0.09). We then tested associations with different clinical and laboratory features, finding that, as expected, MGUS associated with higher concentration of gamma-globulins (Wilcoxon test, p-val = 0.01414), but also with absolute lower levels of albumin (Wilcoxon test, p-val = 0.01398). No significant association was found with the mean corpuscular value (MCV), hemoglobin levels or age. These results were confirmed by a logistic univariate model, in which also the male gender resulted associated with the presence of a monoclonal component with borderline significance. Then, to further corroborate our results we performed univariate linear regression analyses. Given that CHIP and MGUS are mainly considered two aging conditions, we investigated in linear regression models the impact of age, as a continuous variable, on other clinical features. In particular, we observed that the increase of age was significantly correlated with lower albumin levels (F-stat: 1.921e+04, Adj R-squared: 0.9288, p-val: < 2.2e-16), decreasing hemoglobin concentration (F-stat: 1.434e+05, Adj R-squared: 0.9898, p-val < 2.2e-16), increasing levels of gamma-globulins (F-stat: 9.21e+04, Adj R-squared: 0.9843, p-val < 2.2e-16) and higher MCV (F-stat: 18.04, Adj R-squared: 0.01144, p-val 2.3e-05). Then, a significant anti-correlation between gamma-globulin and serum albumin levels was also confirmed (F-stat: 2.807e+04, Adj R-squared: 0.9501, p-val < 2.2e-16). Finally, we implemented two different multivariate logistic models using as independent variables the presence or absence of MGUS or CHIP, respectively. In these models, the presence of a monoclonal component was only positively associated with a higher level of gamma-globulins (Est: 0.16238, p-val = 0.00079) and not with increasing age (Est: 0.02680, p-val = 0.31819). Interestingly, the presence of an MGUS confirmed a tendency to an anti-correlation with the presence of CHIP (Est: -0.78959, p-val = 0.06468). On the contrary, CHIP as independent variable resulted significantly correlated with increasing age (Est: 0.07396, p-val = 0.00021). The spectrum of mutations in CHIP cases with or without MGUS was not significantly different, with DNMT3A and TET2 being the most frequently mutated genes. Our study shows that, in a large cohort of oldest-old patients, CHIP and MGUS are not correlated but follow two seemingly independent patterns, showing a tendency to a mutual exclusivity and associating with different clinical and laboratory values. One limitation of our study is the skewing towards female sex, where MGUS is less prevalent, but this is explained by the cohort's median age knowing that females have a longer expected life. Based on this, our findings that CHIP but not MGUS increased with age in our cohort suggest different selective pressures in this extreme age range. These results warrant further investigation as to whether there could be age-specific drivers for MGUS, and their clinical relevance. Disclosures Bolli: Amgen: Honoraria; Takeda: Honoraria; Janssen: Consultancy, Honoraria; Celgene/BMS: Consultancy, Honoraria.

The thermodynamic Model "Gibbs Free Energy Gradient Method" (GMM), published on the Vol. 90 (2011) of this Journal and validated with literature data, is now applied to the simulation of an experimental campaign realized at the ENEA Research Centre of Trisaia (Italy). The GMM well reproduces the experimental results of steam gasification of refuse-derived fuel (RDF) obtained on two laboratory and pilot scale rotary kilns. Consequently, the experimental syngas composition is put in relation to the main process parameters through a new approach incorporating the GMM for identifying a reliable correlation between the extent of reactions and the gasifier temperature. This correlation appears independent from the scale of the rotary kiln and the residence time in the investigated range of variables. On this basis, the GMM is adapted to become a tool for designing industrial gasifiers starting from experimental data since the required final composition of the syngas and the required performances may be obtained by designing a gasification zone operating at the temperature calculated by the proposed method. It is believed that this procedure is extendable to other geometries and different type of apparatus by studying and including the effect of other parameters. © 2016 Elsevier Ltd. All rights reserved.

Objective: To study dogs bearing otitis externa, objectifying the detection of predisposing factors and microorganisms involved in the etiopathogenesis of this disease. Material and Method: Forty animals (30 of them had otitis externa and 1O without the disease) from the "Servico de Clinica Medica da Faculdade de Medicina Veterinaria da Universidade Paulista", (Medical Clinic Service from the Veterinary Hospital of Paulista Veterinary Medicine University) Sao Paulo, SP, Brazil, were submitted lo prior physical otological examination. The ear secretion samples were obtained from external ear canal using gynecological brushes, submitted to posterior cytological analysis with the help of three different coloration techniques. Results: Among the patients with otitis, 60% was pure breed dogs, 60% female, 74% aged four years old onward, 60% had short-hair coat and 80% with pendulous ears. Gram method was shown as the most effective in the research of bacteria and fungus, and Papanicolau and Rosenfeld techniques were the most adequate to the observation of ephitelial and inflammatory cells and erythrocytes. The most common microorganism found in the extensions produced by the affected ears was Malassezia sp (63.3%). Conclusions: The results indicate that data collected from anamnesis and physical examination, jointly with information provided by the cytological study of ear secretions are efficient in characterizing the elements incriminated in the etiopathogenesis of canine otitis.

Background Determinants of intrafamilial HCV transmission are still being debated. The aim of this study is to investigate the correlates of HCV seropositivity among familial contacts of HCV positive patients in Italy. Methods A cross-sectional study was conducted with 175 HCV positive patients (index cases), recruited from Policlinico Gemelli in Rome as well as other hospitals in Central Italy between 1995 and 2000 (40% female, mean age 57 ± 15.2 years), and 259 familial contacts. Differences in proportions of qualitative variables were tested with non-parametric tests (χ2, Yates correction, Fisher exact test), and a p value < 0.05 was considered significant. A multivariate analysis was conducted using logistic regression in order to verify which variables statistically have an influence on HCV positivity in contact individuals. Results Seropositivity for HCV was found in 8.9% of the contacts. From the univariate analysis, risk factors significantly associated to HCV positivity in the contacts were: intravenous drug addiction (p = 0.004) and intercourse with drug addicts (p = 0.005). The only variables associated significantly and independently to HCV seropositivity in patients' contacts were intercourse with drug addicts (OR = 19.28; 95% CI: 2.01 – 184.94), the retirement status from work (OR = 3.76; 95% CI: 1.17 – 11.98), the time of the relationship (OR = 1.06; 95% CI: 1.00 – 1.11) and tattoos (OR = 7.68; 95% CI: 1.00 – 60.20). Conclusion The present study confirms that having intercourse with a drug addict is the most significant risk factor for intrafamilial HCV transmission. The association with retirement status from work could be related to both a long-term relationship with an index case and past exposure to common risk factors.

There are difficulties in liver surgical procedures, specifically hemostasis, due to anatomic and functional characteristics of this organ. Considering these factors, this study compared the effect of two surgical adhesives, gelatin-resorcin-formaldeíde and n-butil-2-cianoacrilate, in the liver synthesis. Clinical evolution of the animals utilized in this study and cicatrization of experimental liver surgical wounds were investigated. Cicatrization was analysed by macroscopic and microscopic evaluation. Results demonstrated fast and efficient hemostasis in both groups, and more important cellular and tissue alterations on the animals treated by n-butil-2-cianoacrilate, when this group was compared to the control and gelatin-resorcin-formaldeíde groups. Considering the purpose of this study, gelatin-resorcin-formaldeíde compound was the best alternative method on cases where the sutures employed alone had failed to provide adequate hemostasis. Considerando-se as particularidades anatômicas e funcionais do fígado e a dificuldade da realização de procedimentos cirúrgicos eficientes quanto a hemostasia, estudou-se comparativamente os efeitos de dois adesivos cirúrgicos, gelatina-resorsina-formaldeído e n-butil-2-cianoacrilato, como métodos auxiliares na síntese do órgão. A metodologia utilizada determinou rigoroso acompanhamento clínico dos animais estudados, assim como realização de análises macro e microscópica de feridas cirúrgicas experimentalmente induzidas, quanto à evolução do processo cicatricial. Os resultados obtidos demonstraram hemostasia rápida e eficiente em ambos grupos, e alterações celulares e teciduais mais significativas no grupo tratado com n-butil-2-cianoacrilato, quando comparadas àquelas observadas nos grupos controle e gelatina-resorcina-formaldeído. Em relação à proposição estudada, o composto gelatina-resorsina-formaldeído apresentou-se como método alternativo de escolha a ser utilizado nos casos cujas suturas aplicadas isoladamente não proporcionem hemostasia satisfatória.

Summary The authors describe in this paper the occurrence of five cases of dioctophymiasis in Maned wolves Chrysocyon brachyurus belonging to Sao Paulo Zoo. Clinical symptoms are described and the lesions found at post-mortem examination are noted. Agreeing with the veterinary literature on the subject, they also verify that the infestation is in the right kidney. Finally, they show the possibility of diagnosis by radiography and treatment by nephrectomy.

In the x-irradiated mammal, the ratio erythrocytes/nucleated cells is positively correlated with the radiation dose. In the present report, this correlation is used to estimate, by means of marrow biopsies, the radiation dose received by whole-body exposed monkeys.

Os autores relatam a realizacao de fistula experimental canulada do rumen em 10 animais da especie bovina. A tecnica da rumenostomia foi efetuada em apenas um ato operatorio, com os animais em estacao, utilizando-se a anestesia local infiltrativa e com fixacao do rumen na parede muscular do abdomen. O procedimento mostrou-se satisfatorio, com perfeita adaptacao da cânula, formacao de aderencia completa do rumen e parede abdominal e continuidade de sua mucosa com o revestimento cutâneo, sem complicacoes pos-operatorias.

A simplified surgical technique for ear cropping was studied in 120 dogs. The technique consisted of trimming of the ears after clamping, without the use of cauterization nor sutures. The wounds exhibited a faultless healing in a short time and few imperfections were observed at the ears. The cosmetic aspect was satisfatory.

The measurements and weight characteristics of the excised auricular pieces at standardized ear cropping were observed in 24 Doberman Pincher dogs. Good aesthetical aspects were observed in different ages.

Sao estudados em 30 caes, os resultados obtidos com a aplicacao endovenosa de cloridrato de xilazina a 2% seguida da administracao de pentobarbital sodico a 3%. Sao comparados os resultados com os obtidos em um lote de 5 animais anestesiados exclusivamente com pentobarbital sodico e observados, nos lotes experimentais, reducao da quantidade do barbiturico consumido e dos tempos medios de duracao da fase de anestesia cirurgica e necessario para o retorno. As frequencias de pulso e respiratoria mostraram-se diminuidas logo apos a aplicacao das 2 drogas. No tocante a temperatura retal apenas os animais do lote controle apresentaram diferencas significativas ao nivel de 0,05.

Sao estudados em 6 animais da especie canina, tecnicaoperatoria para tratamento da hernia perineal. Sao empregadas paraoclusao da porta hemiaria, malha de polipropileno fixada ao tecidomuscular circundante por meio de sutura em pontos separados comfio numero 00 do mesmo material. Obtem-se resultados satisfaroriosem todos animais operados que mostraram recuperacao completa,sem ocorrencia de alteracoes macroscopicas.

Anesthesia by endovenous injection of 1,01 to 1,50mg/kg of xylazine followed by administration of different doses of pentobarbital sodium were studied in 25 dogs. It was observed different times of animal surgical anesthesia as so in the necessary times to recover and those verified in 5 control dogs. Statistical analysis revealed significant differences (P < .05) among control and treated groups.

The authors study in 9 animals of the bovine species, Holstein Friesian, the surgical technique to correct the umbilical hernia. To obstruct the hernial ring they use separate points in "U" (lateral overlapping technique) with a thread of polypropylene no2. After they reinforce the suture applying polypropylene mesh on the sheet of rectus m. and then suture it using separate points and thread of the same material. They obtain good results in all the operated animals. They recovered completely and there were no macroscopic alterations. After 110 days of the polypropylene applications there was a histopatologic exam in one of the animals. This exam showed a little chronicle inflammatory process in the tissue that involved the polypropylene mesh.

The behavior of 60 surgical patient dogs anesthetized endovenously with 1,01 - 1,50 mg/kg of xylazine followed by sodium pentobarbital as necessary for operation types, were studied. Groups of dogs were stablished based on both operation length and barbituric consumption. It was observed in 37 dogs (61,66%) a consumption of 5,09 mg/kg of pentobarbital/17,43 minutes of operation; 19 animals (31,66%) were operated with 10,27 mg/kg of pentobarbital in 30,26 minutes; 3 dogs (5,00%) used 15,02mg/kg in 48,33 minutes. In only one case it was necessary 18,92mg/kg for a period of 70 minutes.

Foi estudada, em 8 animais da especie canina, uma tecnica operatoria para correcao de defeitos do pavilhao auricular, mediante implantacao de malha de polipropileno. Os resultados foram satisfatorios em 7 caes, que mostraram posicao ereta e movimentacao adequada das orelhas e ausencia de retracoes ou espessamento dos pavilhoes auriculares.

The use of Kuntscher medullary nailing with exposure of femur fracture field was studied in 9 dogs. From the fracture site, a guide pin was driven upward into the trochanteric region. Over the guide pin, the Kuntscher nail was driven downward from the trochanteric fossa. After the normal bony consolidation, the nail was removed. A solid fixation was provided controlling rotation as well as axial displacement by the “V” shaped cross section of the medullary nail.

Injecoes de 0,5 mg de prostaglandina F 2 alfa foram aplicadas, por via subcutânea, em 40 cadelas com sintomas clinicos de pseudociese. O produto mostrou-se eficiente em 37 animais, havendo necessidade de repeticao do tratamento em 4 casos. Em cerca de7 a 8 dias apos o tratamento houve retorno ao comportamento normal. Efeitos colaterais como vomitos e anorexia foram observados em um animal e sonolencia em outros tres. Discute-se o efeito luteolitico da PGF 2 alfa com base na regressao dos sintomas, embora nao se disponha de metodos adequados para analise quantitativa de LH e progesterona.

The authors reported a now method of trating so called hoematoma affecting dog’s ear, using in all cases an injection of triamcinolone acetonide (10 mg/ml), and all the fluid contained was removed prior the injection. After looking over the literature concerning and discussing the results obtained, the authors were able to conclude: The value of treatment with the synthetic corticosteroid injection has definitively been established and has given satisfactory results in at least 83,3 percent of the cases treated; 52 percent of these, have hand only one injection, 40 percent two and 8 percent three injections. In all cases the injections were done every 7 days. They didn't verify the thickness of the ear in 52 percent of the cases treated.