Failure of Miltefosine Treatment in Two Dogs with Natural Leishmania infantum Infection

Two dogs, with naturally acquired canine leishmaniasis, were treated orally with miltefosine (2 mg/kg q 24 hr) and allopurinol (10 mg/kg q 12 hr) for 28 days. Both dogs showed good initial response to therapy, with reduction in clinical signs and improvement of clinicopathological changes. However, in both dogs, clinical and clinicopathological abnormalities recurred 150 days after initial treatment and a second course of miltefosine and allopurinol was administered. One dog failed to respond to the 2nd cycle of miltefosine treatment and the other dog responded initially but suffered an early relapse. Treatment with meglumine antimoniate (100 mg/kg q 24 hr for a minimum of 4 weeks) was then started in both dogs. Both dogs showed rapid clinical and clinicopathological improvement and to date they have not received further treatment for 420 and 270 days, respectively. In view of the low number of antileishmanial drugs available and the fact that some of these are used in human as well as veterinary medicine, it is of paramount importance that drug resistance is monitored and documented

Ammonia concentration and bacterial evaluation of feline whole blood and packed red blood cell units stored for transfusion

Ammonia concentrations increase in human, canine and equine WB and PRBC units during storage. The aim of this study was to determine the effect of storage on ammonia concentration in feline WB and PRBC units stored in a veterinary blood bank and to evaluate possible correlations with bacterial contamination. Ammonia concentration was evaluated in 15 WB units and 2 PRBC units on day 1 and at the end of storage after 35 and 42 days, respectively. In an additional 5 WB units and 4 PRBC units ammonia concentrations were determined daily until the day the normal reference range was exceeded and then weekly to the end of storage. All units were evaluated for bacterial contamination. Ammonia increased markedly during storage as a linear function over time. On the 35th and 42th day of storage at 4±2°C mean±SD ammonia concentration reached 909±158 µg/dl and 1058±212 µg/dl in WB and PRBC units, respectively. Bacterial culture was negative in all units. High ammonia concentrations in stored WB and PRBC units could result in toxicity, particularly in feline recipients with liver failure, portosystemic shunts or those receiving large transfusion volumes. Clinical in vivo studies evaluating the effects on recipients should be performed

FELINE LEISHMANIASIS: SEROLOGICAL AND MOLECULAR DETECTION OF AN EMERGENT DISEASE IN A NON-ENDEMIC AREA OF NORTHERN ITALY

In recent decades feline leishmaniosis (FeL) has become an emerging disease, also in non-endemic areas for the canine infection. This study updates the epidemiological status for FeL in cats in northern Italy and compares results with previous studies of the same feline population. Co-infections with feline retroviruses FIV and Field were also investigated. Stray, shelter and owned cats from different cities in the Lombardy region of northern Italy, were prospectively randomly sampled between January 2020 and May 2021. A total of 255 cats were tested for L. infantum: 240/255 for antibodies by IFAT and 234/255 and 198/255 for Leishmania DNA by PCR on whole blood and lymph nodes, respectively. Rapid ELISA test was used to detect FIV or FeLV infection. Overall, 26/255 (10.2%) cats tested positive for L. infantum: in 8/26 cats Leishmania DNA was found in popliteal lymph nodes (Leishmania/ml range from 15 to 60), 6/26 were PCR positive on whole blood (Leishmania/ml range from 5 to 80) and 15/26 IFAT seropositive at titers ranging from 1:80 to 1:320. Two Leishmania infected cats were also FIV+FeLV coinfected, another was FIV positive and one was FeLV positive. A high prevalence of FeL was found in a non-endemic area of northern Italy, with an increasing trend in infection rates

Comparison of a clinic-based ELISA test kit with the immunofluorescence antibody test for assaying Leishmania infantum antibodies in dogs

This study compares a rapid Immunospecific Kalazar Canine Rapid Spot IF with the gold standard test (indirect fluorescent antibody test (IFAT)) for detection of Leishmania infantum specific IgG serum antibodies in naturally exposed dogs. Serum samples were obtained from 89 healthy dogs and dogs affected by canine leishmaniosis (CanL). IgG-IFAT titers ≥80 were considered positive. Anti-L. infantum IgG antibodies were found in 54 samples with titers ranging from 1 : 80 to 1 : 5120. The performance of the rapid Immunospecific Kalazar was evaluated using a ROC curve. The area under the ROC curve of 0.957 was significantly different from 0.5 ( < 0.0001), and therefore it can be concluded that the rapid Immunospecific Kalazar has the ability to distinguish canine sera with and without L. infantum IgG. The best performance of the test was at a cutoff >0 (sensitivity 92.6%, specificity 97%). The test can be used for disease screening if the cutoff is >0 (highest sensitivity, 92.6%) and is recommended as confirmatory test for the presence of L. infantum IgG antibodies if the cutoff is set >2 (highest specificity, 100%)

Frecuencia del grupo sanguíneo A, B y AB y riesgo de transfusión en gatos del Norte de Italia

Objetivos Datos epidemiol\uf3gicos sobre el tipo de sangre felina son importantes para prevenir reacciones de transfu- si\uf3n y para identificar los donantes de sangre ideales en programas de donantes de sangre felina. Tres eran los objetivos de este estudio: 1. Determinarlafrecuenciadelosgrupossangu\uedneos en una poblaci\uf3n de gatos con tipaje sangu\uedneo en la Universidad de Mil\ue1n 2. Analizar las caracter\uedsticas de la poblaci\uf3n y pro- bar la asociaci\uf3n entre el tipo de sangre A, B y AB y una serie de variables (sexo, raza y grupo filoge- n\ue9tico); 3. Calcular el riesgo de reacci\uf3n a la transfusi\uf3n menor y mayor despu\ue9s de la administraci\uf3n de sangre incompatible despu\ue9s de una transfusi\uf3n al azar en esta poblaci\uf3n. 4. Calcular el riesgo de una reacci\uf3n a latransfusi\uf3n mayor o menor despu\ue9s de una administraci\uf3n de sangre incompatible de forma aleatoria en esta poblaci\uf3n Material y m\ue9todos Un estudio retrospectivo se realiz\uf3 utilizando datos (recogidos entre 2010 y 2014) de la base de datos de la unidad de transfusi\uf3n veterinaria (REV) de la Universidad de Mil\ue1n, Italia. Los datos recogidos inclu\uedan: tipo de sangre, edad, sexo, raza, raz\uf3n y el m\ue9todo de determinaci\uf3n del grupo sangu\uedneo. Los resultados se analizaron por an\ue1lisis de frecuencia absoluta y relativa. Las frecuencias de los diferentes grupos sangu\uedneos y datos categ\uf3ricos (sexo, raza y grupo filogen\ue9tico) se compararon mediante tablas de contingencia y la prueba exacta de Fisher o prueba de chi cuadrado, seg\ufan el caso. Se seleccionaron Conclusiones 88 gatos como potenciales donantes de sangre, el grupo sangu\uedneo y otros factores (precruce, preoperatorio y receptor) fueron considerados aptos para el objetivo de los c\ue1lculos. La posibilidad de una reacci\uf3n mayor de transfusi\uf3n en gato con sangre del grupo B que recibe sangre del tipo A o AB, se calcul\uf3 de la siguiente manera: frecuencia del donante tipo A + frecuencia d\uf3nate tipo AB multiplicada ambas por frecuencia de receptor B. La posibilidad de una reacci\uf3n de transfusi\uf3n menor (la reacci\uf3n menor reduce la vida media de los eritrocitos transfundidos en sangre de gatos del grupo sangu\uedneo A que recibe sangre del grupo sangu\uedneo B o AB y gatos del grupo sangu\uedneo AB que reciben sangre de grupo B) fue calculada usando la f\uf3rmula: frecuencia del gato donante tipo B donante + frecuencia del donante tipo AB multiplicado por la frecuencia del receptor tipo A + frecuencia donante tipo B multiplicado por frecuencia receptor tipo AB (Juvet F et al 2011). Resultados 282 fueron incluidos en el estudio, el rango de edad fue desde 2 meses a 19 a\uf1os, 140 (50%) macho y 138 (49%) hembras, 151 (53%) gato com\ufan europeo de pelo corto, 59 (21%) Ragdoll, 29 (10%) Maine Coons. Los gatos se dividieron en tres grupos de acuerdo con su origen 55% de los gatos eran de la cuenca medite- rr\ue1nea, 44% del oeste de Europa, 1% de Asia y no se tipificaron gatos de \uc1frica oriental. La prevalencia de los grupos sangu\uedneos fue 91% (n= 257) tipo A, 4% (n=11) tipo B y 5% (n=14) tipo AB. Las razones para realizar el grupo sangu\uedneo fueron: precruce para pre- venir Isoeritrolisis neonatal en 92 gatos (33%), scree- ning preoperatorio en 55 gatos (19%), examen pre- transfusi\uf3n en 17 gatos (6%) y screening de potenciales donantes previo a la donaci\uf3n en 88 gatos (31%). No hubo asociaci\uf3n significativa entre el grupo sangu\uedneo y la variable analizada, excepto por una asociaci\uf3n entre gato Ragdoll y grupo sangu\uedneo A (83%) y AB (14%) (P=0,0335, OR=0,3 y P=0,0026, OR=5,6 respecti- vamente). Todos los gatos Maine Coon testado fueron grupo sangu\uedneo A. En una transfusi\uf3n de sangre sin testar la frecuencia estimada de una reacci\uf3n mayor y menor fue 3.8% y 9.3% respectivamente. Discusi\uf3n De acuerdo con los hallazgos de otros estudios europeos (Jensen AL et al, 1994; Knottembelt CM et al 1999; Ruiz de Gopegui R et al 2004; Marques C et al 2011; Juvet F et al, 2011), el grupo sangu\uedneo predominante en los gatos en este estudio fue de tipo A. La prevalencia del grupos sangu\uedneo var\ueda entre las diferentes razas felinas. El tipo de sangre AB es significativamente m\ue1s frecuente en los gatos de Ragdoll que en otras razas feli- nas y este hallazgo podr\ueda ser \ufatil cuando se consideran candidatos para la donaci\uf3n de plasma. Conclusiones Dado que el riesgo de una reacci\uf3n transfusional en los gatos que recibieron una transfusi\uf3n de sangre sin la comprobaci\uf3n de la compatibilidad sangu\uednea que resulto ser del 9% el grupo sangu\uedneo y las pruebas cruzadas de los donantes de sangre y el receptor son esenciales antes de la transfusi\uf3n felina para prevenir la potencial reacci\uf3n transfusional inmunol\uf3gica o la r\ue1pida destrucci\uf3n de los eritrocitos transfundidos. Bibliograf\ueda \u2022 jensen AL, Olesen AB, Arnbjerg J. Distribution of feli- ne blood types obtained in the Copenhagen area of Den- mark. Acta Vet Scand. 1994;35:121-4. \u2022 Knottenbelt CM, Addie DD, Day MJ, Mackin AJ. Deter- mination of the prevalence of feline blood types in the UK. J Small Anim Pract. 1999;40:115-8. \u2022 Ruiz de Gopegui R, Velasquez M, Espada Y. Survey of fe- line blood types in the Barcelona area of Spain. Vet Rec. 2004;154:794-5. \u2022 Marques C, Ferreira M, Gomes JF. Frequency of blood type A, B and AB in 515 domestic shorthair cats from the Lisbon area. Vet Clin Pathol. 2011;40:185-7. \u2022 Juvet F, Brennan S, Mooney CT. Assessment of feline blood for transfusion purposes in the Dublin area of Ireland. Vet Rec. 2011;168:352-4

Octreotide Conjugates for Tumor Targeting and Imaging

Tumor targeting has emerged as an advantageous approach to improving the efficacy and safety of cytotoxic agents or radiolabeled ligands that do not preferentially accumulate in the tumor tissue. The somatostatin receptors (SSTRs) belong to the G-protein-coupled receptor superfamily and they are overexpressed in many neuroendocrine tumors (NETs). SSTRs can be efficiently targeted with octreotide, a cyclic octapeptide that is derived from native somatostatin. The conjugation of cargoes to octreotide represents an attractive approach for effective tumor targeting. In this study, we conjugated octreotide to cryptophycin, which is a highly cytotoxic depsipeptide, through the protease cleavable Val-Cit dipeptide linker using two different self-immolative moieties. The biological activity was investigated in vitro and the self-immolative part largely influenced the stability of the conjugates. Replacement of cryptophycin by the infrared cyanine dye Cy5.5 was exploited to elucidate the tumor targeting properties of the conjugates in vitro and in vivo. The compound efficiently and selectively internalized in cells overexpressing SSTR2 and accumulated in xenografts for a prolonged time. Our results on the in vivo properties indicate that octreotide may serve as an efficient delivery vehicle for tumor targeting.Peer reviewe

Differential expression of microRNAs between eutopic and ectopic endometrium in ovarian endometriosis

Endometriosis, defined as the presence of endometrial tissue outside the uterus, is a common gynecological disease with poorly understood pathogenesis. MicroRNAs are members of a class of small noncoding RNA molecules that have a critical role in posttranscriptional regulation of gene expression by repression of target mRNAs translation. We assessed differentially expressed microRNAs in ectopic endometrium compared with eutopic endometrium in 3 patients through microarray analysis. We identified 50 microRNAs differentially expressed and the differential expression of five microRNAs was validated by real-time RT-PCR in other 13 patients. We identified in silico their predicted targets, several of which match the genes that have been identified to be differentially expressed in ectopic versus eutopic endometrium in studies of gene expression. A functional analysis of the predicted targets indicates that several of these are involved in molecular pathways implicated in endometriosis, thus strengthening the hypothesis of the role of microRNAs in this pathology

Serum Albumin Is Inversely Associated With Portal Vein Thrombosis in Cirrhosis

We analyzed whether serum albumin is independently associated with portal vein thrombosis (PVT) in liver cirrhosis (LC) and if a biologic plausibility exists. This study was divided into three parts. In part 1 (retrospective analysis), 753 consecutive patients with LC with ultrasound-detected PVT were retrospectively analyzed. In part 2, 112 patients with LC and 56 matched controls were entered in the cross-sectional study. In part 3, 5 patients with cirrhosis were entered in the in vivo study and 4 healthy subjects (HSs) were entered in the in vitro study to explore if albumin may affect platelet activation by modulating oxidative stress. In the 753 patients with LC, the prevalence of PVT was 16.7%; logistic analysis showed that only age (odds ratio [OR], 1.024; P = 0.012) and serum albumin (OR, -0.422; P = 0.0001) significantly predicted patients with PVT. Analyzing the 112 patients with LC and controls, soluble clusters of differentiation (CD)40-ligand (P = 0.0238), soluble Nox2-derived peptide (sNox2-dp; P < 0.0001), and urinary excretion of isoprostanes (P = 0.0078) were higher in patients with LC. In LC, albumin was correlated with sCD4OL (Spearman's rank correlation coefficient [r(s)], -0.33; P < 0.001), sNox2-dp (r(s), -0.57; P < 0.0001), and urinary excretion of isoprostanes (r(s), -0.48; P < 0.0001) levels. The in vivo study showed a progressive decrease in platelet aggregation, sNox2-dp, and urinary 8-iso prostaglandin F2 alpha-III formation 2 hours and 3 days after albumin infusion. Finally, platelet aggregation, sNox2-dp, and isoprostane formation significantly decreased in platelets from HSs incubated with scalar concentrations of albumin. Conclusion: Low serum albumin in LC is associated with PVT, suggesting that albumin could be a modulator of the hemostatic system through interference with mechanisms regulating platelet activation

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements