Highly recurrent BRAF p.V595E mutation in canine papillary oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is the most common oral epithelial malignancy in dogs. It exhibits locally aggressive biological behaviour with the potential to metastasize, and a reported 1-year survival rate of 0% when left untreated. Expression studies suggest that aberrant MAPK signalling plays a key role in canine OSCC tumorigenesis, which is consistent with BRAF and HRAS MAPK-activating mutations reported in some tumours. Several morphological subtypes of canine OSCC have been described, with papillary, conventional, and basaloid as the most common patterns. We hypothesized that mutational differences may underlie these phenotypic variations. In this study, targeted Sanger sequencing and restriction fragment length polymorphism assays demonstrate that up to 85.7% of canine papillary OSCC (n = 14) harbour a BRAF p.V595E mutation. Assessment of neoplastic epithelial cell proliferation using Ki67 immunolabelling (n = 10) confirmed a relatively high proliferation activity, consistent with their known aggressive clinical behaviour. These findings underscore a consistent genetic feature of canine papillary OSCC and provide a basis for the development of novel diagnostic and targeted therapeutic approaches that can improve the quality of veterinary care.     

The screening and evaluation of herbs and identification of herbal combinations with anti-viral effects on Newcastle disease virus

1. A total of 25 “heat-clearing and detoxifying” herbs used in Chinese medicine were investigated for their cytopathic effects on the growth of Newcastle Disease virus (NDV) in a chicken fibroblast cell line.

2. The 5 herbs with the highest virus inhibitory effects were Herba agastaches, Flos chrysanthemi indici, Rhizoma anemarrhenae, Astragalus root and Baikal skullcap root and these were used in herbal formulations. Anti-NDV activities of 4 formulations were tested on the growth of NDV in the DF-1 fibroblast cell line.

3. Formulation II, containing Baikal skullcap root, Astragalus root, Anemarrhena rhizome (1:1:2) and formulation IV containing Anemarrhena rhizome, Astragalus root and Flos chrysanthemi indici (1:1:1), which had strong anti-NDV activity in vitro, were used to determine the in vivo inhibitory effects of NDV-infection in chickens. After treatment with the two formulations serum IgY titres against NDV were improved, and morbidity was reduced in the NDV-infected chickens.

4. The results suggest that the components in formulations II and IV acted synergistically to improve resistance to Newcastle disease and provide a basis for the developing an anti-NDV herbal medicine.

     

Effect of animal age,postmortem chilling rate,and aging time on meat quality attributes of water buffalo and humped cattle bulls

The study was aimed to investigate the influence of animal age, post‐slaughter chilling rate, and aging time on meat quality of M. longissimus dorsi (LD) of water buffalo (Bubalus bubalis) and humped cattle (Bos taurus indicus) bulls. After slaughtering, one side of carcasses was subjected to rapid chilling (RC) (0 ± 2°C) and other side was hanged in controlled room temperature (25 ± 2°C) for 3 hr, then allowed to the chiller (0 ± 2°C). The meat quality traits were analyzed at 1, 7, and 14 days of storage. It was noted that rapidly chilled carcasses from the younger animals of both species missed the ideal pH/temperature window, which affects the toughness of the meat. Buffalo meat presented higher shear force, color L* values, and lower b* value as compared to the cattle meat. Moreover, meat shear force values decreased while all color coordinates and cooking loss values increased with lengthening the storage time in both age groups of cattle and buffalo. In conclusion, the tenderness of cattle meat was superior to that of buffalo and RC adversely affect the shear force values of young cattle and both age groups of buffalo bulls.     

Use of rotational periosteal flaps in primary repair of sinofacial fractures in seven horses

ABSTRACT

Case history: Medical records were reviewed of horses (n?=?7) undergoing surgery for fracture of one or more facial bones extending into the paranasal sinuses that was repaired primarily within 24 hours of the time of injury using a rotational periosteal flap, between April 2009 and May 2017. A kick from another horse was the cause of the injury of three horses, and one horse was injured when it collided with a tree. The cause of the injury of three horses was unknown.

Clinical findings and treatment: Fractures were of the right maxillary bone in two horses, the left maxillary bone in two horses, the left frontal and left nasal bones in two horses, and the right frontal bones in one horse. The fracture of all but one horse was accompanied by an open wound. The fracture of all seven horses was reduced, stabilised, and covered with a rotational, periosteal flap. Surgery was carried out while standing in six horses, and while anesthetised in one horse. All horses had a deficit in the fractured facial bones after the fracture was reduced. Four horses had complications following surgery, but all horses were reported to have excellent cosmetic outcomes and had retuned to their previous level of activity, as reported by their owners.

Clinical relevance: Covering a primarily repaired sinofacial fracture of a horse with a rotational periosteal flap resulted in good cosmetic outcomes, and may be especially beneficial if the fracture is accompanied by loss of bone.     

SV40 Immortalization of feline fibroblasts as targets for MHC-restricted cytotoxic T-cell assays

CTL assays in outbred cats have been difficult to perform because of a lack of a good source of syngeneic target cell. Primary fibroblasts from cats are widely used as target cells for MHC-restricted cytotoxic T-cell (CTL) assays, but their limited life-spans of 8-10 culture passages can be problematic for longitudinal studies. To circumvent the life-span limitations of primary fibroblast cultures, we developed a procedure for immortalizing feline primary fibroblast cells by transfection with a molecular clone of simian virus 40 (SV40). Fibroblast cultures from skin biopsies of 28 cats were immortalized using this procedure and have been passaged for longer than 6 months without showing any phenotypic difference from the original primary cells. Non-SV40 transfected feline fibroblasts from a selection of animals in the same group survived for only 6-8 weeks before reaching senescence. The immortalized fibroblasts expressed SV40 T-antigen and Class I MHC protein, and were successfully used as target cells in 51Cr release CTL assays in feline immunodeficiency virus (FIV)-infected cats and in vitro stimulated allogeneic T-cell cultures.     

Ultrasonography and surgery of canine biliary diseases

Findings of hepatic and gallbladder ultrasonography were analyzed in 12 dogs with gallbladder and/or extrahepatic biliary tract obstruction and compared with the results of exploratory laparotomy. Hepatic ultrasonography demonstrated normal liver in 2 dogs and hepatic abnormalities in 10 animals. The following ultrasonographic diagnoses were established compared to surgical findings: gallbladder obstruction caused by bile sludge (correct/incorrect: 1/2, surgical diagnosis: choleliths in one case), gallbladder obstruction caused by neoplasm (0/1, surgical diagnosis: mucocele), gallbladder and extrahepatic biliary tract obstruction due to choleliths (3/3), extrahepatic biliary tract obstruction caused by pancreatic mass (1/1) and small intestinal volvulus (1/1). Bile peritonitis caused by gallbladder rupture (4/4) was correctly diagnosed by ultrasound, aided with ultrasonographically-guided abdominocentesis and peritoneal fluid analysis. Rupture of the gallbladder should be suspected in the presence of a small, echogenic gallbladder or in the absence of the organ together with free abdominal fluid during ultrasonography. Laparotomy was correctly indicated by ultrasonography in all cases. However, the direct cause of obstruction could not be determined in 2 of the 12 dogs by ultrasonography alone.     

Selection for growth rate or against back fat thickness in pigs is associated with changes in growth hormone axis plasma protein concentration and mRNA level.

Selection for increased growth rate or decreased back fat thickness results in concomitant changes in endocrine and metabolic status. Growth hormone (GH) changes in blood plasma concentration related to selection for growth rate and fat deposition were reported in pigs. The molecular mechanisms regulating selection-induced changes in GH plasma concentration remain largely unknown. We investigated selection-associated changes in GH axis parameters in 2 pig lines selected for increased growth rate (F-line), or decreased back fat thickness (L-line), respectively. First, we investigated selection-associated changes in GH pulse parameters. In both selection lines we found each generation a declining GH peak maximum concentration and area under the GH curve. GH pulse width was not associated with generation number. In both lines generation number was associated with a declined pulse interval, indicating that the number of pulses per day increased on average with 1 pulse per 24 h per generation. Second, plasma concentration of GH axis related Insulin-like growth factor-I (IGF-I) and insulin were investigated. Plasma IGF-I concentration was not associated with generation number in the F-line. Mean plasma insulin concentration declined each generation in both lines. Third, we investigated changes in GH and Pit-1 mRNA levels. In both selection lines GH and Pit-1 mRNA levels increased approximately 50% each generation. The high SD of the GH mRNA levels in both lines may suggest that the GH mRNA levels are pulsatile in vivo. We postulate a molecular mechanism that may explain how selection is associated with increased GH mRNA levels and GH pulse numbers, while lowering GH release per pulse.