Identifying hydric, electrolytic and acid-base imbalances through traditional and quantitative approaches in dogs with hemorrhagic gastroenteritis / Identificação de desequilíbrios hídricos, eletrolíticos e ácido-base pelas abordagens tradicional e quantitativa em cães com gastroenterite hemorrágica

Vomiting and diarrhea are two important clinical signs that can cause significant electrolytic and acid-base imbalances. The purposes of this study were to characterize hydric, electrolytic and acid-base disorders presented by puppies with hemorrhagic gastroenteritis and to compare the traditional and quantitative approaches to acid-base status interpretation. Sixty-one animals with a history of vomiting and/or diarrhea were used in this study and the following tests were performed: complete blood count, total plasma protein concentration and hemogasometry. Mean, standard deviation and Kappa values were calculated. The imbalances characterized by both approaches were: 42 (69%) animals without imbalance, 17 (28%) with metabolic alkalosis and 2 (3%) with metabolic acidosis by the traditional approach and 17 (28%) dogs without imbalance, 26 (43%) with metabolic alkalosis and 18 (29%) with metabolic acidosis by the quantitative approach. The agreement calculated between two approaches coincide in 28 cases, with a moderate Kappa value equivalent to 0.459. The most found imbalances were metabolic alkalosis, hypokalemia, and mild dehydration. Most of acid-base disturbances were not identified by the traditional approach, whereas by the quantitative approach, they were easily determined. Thus quantitative approach proved to be superior in identification of possible acid-base imbalances.(AU)

Vômito e diarreia são dois sinais clínicos importantes, que podem causar desequilíbrios eletrolíticos e ácido-base importantes. Os objetivos deste trabalho foram caracterizar os desequilíbrios hídrico, eletrolítico e ácido-base apresentados por filhotes de cães com gastroenterite hemorrágica e comparar as abordagens de interpretação do equilíbrio ácido-base tradicional e quantitativa. Sessenta e um animais com histórico de vômito e/ou diarreia foram utilizados neste estudo, e os seguintes testes foram realizados: hemograma, concentração de proteína total plasmática e hemogasometria. Média, desvio-padrão e valor de Kappa foram calculados. Os desequilíbrios caracterizados pelas duas abordagens foram: 42 (69%) animais sem desequilíbrio, 17 (28%) com alcalose metabólica e dois (3%) com acidose metabólica, pela abordagem tradicional, e 17 (28%) cães sem desequilíbrio, 26 (43%) com alcalose metabólica e 18 (29%) com acidose metabólica, pela abordagem quantitativa. A concordância calculada entre as duas abordagens coincidiu em 28 casos, com um valor de Kappa moderado, equivalente a 0,459. Os desequilíbrios mais encontrados foram alcalose metabólica, hipocalemia e desidratação leve. A maioria dos distúrbios não foram identificados pela abordagem tradicional, enquanto, pela abordagem quantitativa, foram facilmente determinados. Portanto, a abordagem quantitativa provou-se superior na identificação de possíveis desequilíbrios ácido-base.(AU)

PDX1 -MODY and dorsal pancreatic agenesis: New phenotype of a rare disease.

Maturity-Onset Diabetes of the Young (MODY) type 4 or PDX1 -MODY is a rare form of monogenic diabetes caused by heterozygous variants in PDX1 . Pancreatic developmental anomalies related to PDX1 are reported only in neonatal diabetes cases. Here, we describe dorsal pancreatic agenesis in 2 patients with PDX1 -MODY. The proband presented with diabetes since 14 years of age and maintained regular glycemic control with low doses of basal insulin and detectable C-peptide levels after 38 years with diabetes. A diagnosis of MODY was suspected. Targeted next-generation sequencing identified a heterozygous variant in PDX1 : c.188delC/p.Pro63Argfs*60. Computed tomography revealed caudal pancreatic agenesis. Low fecal elastase indicated exocrine insufficiency. His son had impaired glucose tolerance, presented similar pancreatic agenesis, and harbored the same allelic variant. The unusual presentation in this Brazilian family enabled expansion upon a rare disease phenotype, demonstrating the possibility of detecting pancreatic malformation even in cases of PDX1 -related diabetes diagnosed after the first year of life. This finding can improve the management of MODY4 patients, leading to precocious investigation of pancreatic dysgenesis and exocrine dysfunction.

Clinical application of ACMG-AMP guidelines in HNF1A and GCK variants in a cohort of MODY families.

Maturity-onset diabetes of the young (MODY) is a form of monogenic diabetes with autosomal dominant inheritance. GCK -MODY and HNF1A -MODY are the prevalent subtypes. Currently, there is growing concern regarding the correct interpretation of molecular genetic findings. The American College of Medical Genetics and Genomics (ACMG) updated guidelines to interpret and classify molecular variants. This study aimed to determine the prevalence of MODY ( GCK / HNF1A ) in a large cohort of Brazilian families, to report variants related to phenotype, and to classify them according to ACMG guidelines. One hundred and nine probands were investigated, 45% with clinical suspicion of GCK -MODY and 55% with suspicion of HNF1A -MODY. Twenty-five different variants were identified in GCK gene (30 probands-61% of positivity), and 7 variants in HNF1A (10 probands-17% of positivity). Fourteen of them were novel (12- GCK /2- HNF1A ). ACMG guidelines were able to classify a large portion of variants as pathogenic (36%- GCK /86%- HNF1A ) and likely pathogenic (44%- GCK /14%- HNF1A ), with 16% (5/32) as uncertain significance. This allows us to determine the pathogenicity classification more efficiently, and also reinforces the suspected associations with the phenotype among novel variants.

TcCYS4, a cystatin from cocoa, reduces necrosis triggered by MpNEP2 in tobacco plants.

In Brazil, most cocoa bean production occurs in Southern Bahia. Witches' broom disease arrived in this area in 1989 and has since caused heavy losses in production. The disease is caused by the basidiomycete fungus Moniliophthora perniciosa, a hemibiotrophic fungus that produces the necrosis and ethylene-inducting protein (MpNEP2) during infection; this protein can activate cysteine proteases and induce programmed cell death. Cysteine proteases can be modulated by cystatin. In this study, we overexpressed TcCYS4, a cocoa cystatin, in tobacco plants and evaluated the effect on MpNEP2 in model plants. Tccys4 cDNA was cloned into the pCAMBIA 1390 vector and inserted into the tobacco plants via Agrobacterium tumefaciens. Transgene expression was analyzed by reverse transcription-quantitative PCR and Western blot analysis. Transcript and protein levels in Tcccys4:tobacco lines were 8.9- and 1.5-fold higher than in wild-type plants (wt). Tcccys4:tobacco lines showed no change in growth compared to wt plants. CO2 net assimilation (A) increased in Tcccys4:tobacco lines compared to wt plants. Only one line showed statistically significant stomatal conductance (gs) and transpiration rate (E) changes. MpNEP2 was infiltered into the foliar mesophyll of Tcccys4:tobacco lines and wt plants, and necrotic lesions were attenuated in lines highly expressing Tccys4. Our results suggest that cocoa cystatin TcCYS4 affects MpNEP2 activity related to the progression of programmed cell death in tobacco plants. This may occur through the action of cystatin to inhibit cysteine proteases activated by MpNEP2 in plant tissues. Further studies are necessary to examine cystatin in the Theobroma cacao-M. perniciosa pathosystem.