The number of episodes of diabetic ketoacidosis (DKA) is a significant outcome measure for diabetes care. We ascertained patterns of admission due to DKA over 15 yr to determine whether this indicator of diabetes care had improved in parallel with clinical practices. Between 1 January 1985 and 31 December 1999, 630 admissions were reviewed. We subanalyzed these admissions according to whether the patient was newly diagnosed, had infrequent episodes of DKA (non-relapsers) or had frequent (> or = 2/yr) episodes of DKA (relapsers). Overall there was a slight downward trend in the incidence of DKA admissions over the study period. There was a proportionate increase in the incidence of DKA amongst newly diagnosed patients, with a proportionate decrease in the incidence of DKA seen in relapsers. DKA occurring in non-relapsers remained relatively stable. Adverse clinical events during the admission were relatively uncommon and occurred in all three subgroups. There was no significant difference in HbA1C prior to admission between the relapser and non-relapser groups and there was similarity in the degree of acidosis between all three subgroups. The frequency of significant complications associated with DKA remained unchanged over the study period. Slower rehydration policies were not associated with decreases in either cerebral edema or death rates. DKA remains a significant complication of type 1 diabetes associated with a variety of significant adverse events. Our experience indicates that further efforts to reduce the occurrence of DKA must be focused upon earlier diagnosis and intervention in newly diagnosed patients.

Beta-ketothiolase (T2) deficiency is an inherited disease of isoleucine and ketone body metabolism caused by mutations in the ACAT1 gene. Between 2005 and 2016, a total of 41 patients with T2 deficiency were identified at a medical center in northern Vietnam, with an estimated incidence of one in 190,000 newborns. Most patients manifested ketoacidotic episodes of varying severity between 6 and 18 months of age. Remarkably, 28% of patients showed high blood glucose levels (up to 23.3 mmol/L). Ketoacidotic episodes recurred in 43% of patients. The age of onset, frequency of episodes, and identified genotype did not affect patient outcomes that were generally favorable, with the exception of seven cases (five died and two had neurological sequelae). Custom-tailored acute and follow-up management was critical for a positive clinical outcome. Two null mutations, c.622C>T (p.Arg208*) and c.1006-1G>C (p.Val336fs), accounted for 66% and 19% of all identified ACAT1 mutant alleles, respectively. Most patients showed characteristic biochemical abnormalities. A newborn screening program could be expected to have a high yield in Vietnam. Investigation findings of haplotypes linked to the most common ACAT1 mutation (c.622C>T) are consistent with an ancient common founder of mutation-bearing chromosomes belonging to the Kinh ethnic population. The direct management and long-term follow-up of a large number of T2-deficient patients enabled us to study the natural history of this rare disease.

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Raffinose family oligosaccharides (RFOs) are major soluble carbohydrates in soybean seeds that cannot be digested by human and other monogastric animals. Hence, a major goal is to reduce RFO levels to improve the nutritional quality of soybean. In this study, we utilized a dual gRNAs CRISPR/Cas9 system to induce knockouts in two soybean galactinol synthase (GOLS) genes, GmGOLS1A and its homeolog GmGOLS1B. Genotyping of T0 plants showed that the construct design was efficient in inducing various deletions in the target sites or sequences spanning the two target sites of both GmGOLS1A and GmGOLS1B genes. A subset of induced alleles was successfully transferred to progeny and, at the T2 generation, we identified null segregants of single and double mutant genotypes without off-target induced mutations. The seed carbohydrate analysis of double mutant lines showed a reduction in the total RFO content of soybean seed from 64.7 mg/g dry weight to 41.95 mg/g dry weight, a 35.2% decrease. On average, the stachyose content, the most predominant RFO in soybean seeds, decreased by 35.4% in double mutant soybean, while the raffinose content increased by 41.7%. A slight decrease in verbascose content was also observed in mutant lines. Aside from changes in soluble carbohydrate content, some mutant lines also exhibited increased protein and fat contents. Otherwise, no difference in seed weight, seed germination, plant development and morphology was observed in the mutants. Our findings indicate that GmGOLS1A and GmGOLS1B contribute to the soybean oligosaccharide profile through RFO biosynthesis pathways, and are promising targets for future investigation, as well as crop improvement efforts. Our results also demonstrate the potential in using elite soybean cultivars for transformation and targeted genome editing.

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Acute pancreatitis (AP) is acute inflammation of the pancreas, mainly caused by gallstones and alcohol, driven by changes in communication between cells. Heparin-binding proteins (HBPs) play a central role in health and diseases. Therefore, we used heparin affinity proteomics to identify extracellular HBPs in pancreas and plasma of normal mice and in a caerulein mouse model of AP. Many new extracellular HBPs (360) were discovered in the pancreas, taking the total number of HBPs known to 786. Extracellular pancreas HBPs form highly interconnected protein-protein interaction networks in both normal pancreas (NP) and AP. Thus, HBPs represent an important set of extracellular proteins with significant regulatory potential in the pancreas. HBPs in NP are associated with biological functions such as molecular transport and cellular movement that underlie pancreatic homeostasis. However, in AP HBPs are associated with additional inflammatory processes such as acute phase response signalling, complement activation and mitochondrial dysfunction, which has a central role in the development of AP. Plasma HBPs in AP included known AP biomarkers such as serum amyloid A, as well as emerging targets such as histone H2A. Other HBPs such as alpha 2-HS glycoprotein (AHSG) and histidine-rich glycoprotein (HRG) need further investigation for potential applications in the management of AP. Pancreas HBPs are extracellular and so easily accessible and are potential drug targets in AP, whereas plasma HBPs represent potential biomarkers for AP. Thus, their identification paves the way to determine which HBPs may have potential applications in the management of AP.

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Melia azedarach L. is a multipurpose timber tree that is important in forestry plantations. Genetic modification to improve the growth rate and wood quality is important to produce new cultivars of M. azedarach. In this study, the effects of AtGA20ox gene expression on plant growth, wood formation, and biomass production in transgenic M. azedarach plants were investigated. AtGA20ox was inserted into the binary vector pBI101 under the control of the xylem specific promoter CAD4 and transferred to M. azedarach via Agrobacterium-mediated method. The integration and expression of the transgenes were confirmed using PCR, RT-PCR, and Southern blotting. The specificity of CAD4 in the xylem was indicated by the higher expression of the AtGA20ox gene in the stem tissues compared to the leaves. The larger stem diameter resulted from higher xylem cell numbers and a wider xylem zone in all the transgenic plants. In addition, the transgenic plants also grew more quickly. Consequently, transgenic M. azedarach increased its biomass production 2-to 4-fold compared to the wild-type plants at 90 days under greenhouse conditions. Our results suggest that the expression of ectopic AtGA20ox under the control of a xylem-specific promoter is an appropriate method to improve plant growth, biomass production, and wood formation in M. azedarach and other forestry plants.