Peculiar properties of tuber starch in a potato mutant lacking the α-glucan water dikinase 1 gene GWD1 created by targeted mutagenesis using the CRISPR/dMac3-Cas9 system.

Glucose chains in starch are phosphorylated and contribute to structural stabilization. Phosphate groups contained in starch also play a role in retaining moisture. α-Glucan water dikinase 1 (GWD1) is involved in the phosphorylation of glucose chains in starch. In this study, we generated potato mutants of the GWD1 gene using the CRISPR/dMac3-Cas9 system. Observation of the phenotypes of the GWD1-deficient mutants revealed their physiological roles in tuber starch formation. The 4-allele mutants showed growth retardation and a delay in tuber formation. A significant decrease in phosphorus content was detected in the tuber starch of the gwd1 mutant. This mutant starch showed a higher amylose content than the wild-type starch, whereas its gelatinization temperature was slightly lower than that of the WT starch. The peak viscosity of the mutant starch was lower than that of the WT starch. These observations revealed that the starch of the gwd1 mutants had peculiar and unique properties compared to those of WT, sbe3 and gbss1 mutant starches. The amount of tissue-released water due to freeze-thawing treatment was determined on tubers of the gwd1 mutant and compared with those of WT and the other mutants. Significantly less water loss was found in the gwd1, sbe3 and gbss1 mutant tubers than in the WT tubers. Our results indicate that the GWD1 gene is not only important for potato growth, but also largely effective for the traits of tuber starch.

Partial stapled hemorrhoidopexy: clinical aspects and impact on anorectal physiology. / Hemorroidopexia por grampeamento parcial: aspectos clínicos e impacto sob a fisiologia anorretal.

OBJECTIVE:: to evaluate the impact of partial stapled hemorrhoidopexy on anorectal physiology, the complications related to this surgical technique, pain, postoperative bleeding and recurrence of hemorrhoidal disease one year after surgery. METHODS:: this is a prospective, descriptive study in consecutive patients with mixed or internal hemorrhoidal disease, the internal component being classified as grade III or IV, undergoing partial stapled hemorrhoidopexy. RESULTS:: we studied 17 patients, 82% of them with internal hemorrhoids grade III and 18% grade IV. The mean operative time was 09:09 minutes (07:03 to 12:13). The median pain in the immediate postoperative period evaluated by the numerical pain scale was one (0 to 7). The median time to return to work was nine days (4 to 19). No patient had anal stenosis and 76% were satisfied with the surgery 90 days postoperatively. When comparing the preoperative manometry data with that measured 90 days after surgery, none of the variables studied showed statistically significant difference. There was no recurrence of hemorrhoidal disease with one year of postoperative follow-up. CONCLUSION:: partial stapled hemorrhoidopexy showed no impact on anorectal physiology, presenting low levels of complications and postoperative pain, without recurrence of hemorrhoidal disease in one year of follow-up.

Hemorroidopexia por grampeamento parcial: aspectos clínicos e impacto sob a fisiologia anorretal / Partial stapled hemorrhoidopexy: clinical aspects and impact on anorectal physiology

RESUMO Objetivo: avaliar o impacto na fisiologia anorretal da hemorroidopexia por grampeamento parcial, das complicações relacionadas à técnica cirúrgica, dor e sangramento pós-operatório e recidiva de doença hemorroidária após um ano de cirurgia. Métodos: estudo prospectivo, descritivo, em pacientes consecutivos, portadores de doença hemorroidária do tipo mista ou interna, com componente interno classificado como grau III ou IV, submetidos à hemorroidopexia por grampeamento parcial. Resultados: foram estudados 17 pacientes, dos quais 82% apresentavam hemorroidas internas grau III, e 18% grau IV. A média de tempo operatório foi de 09:09 minutos (07:03 a 12:13 minutos). A mediana de dor no pós-operatório imediato avaliada pela escala numérica de dor foi de 1 (0 a 7). A mediana de retorno ao trabalho foi de nove dias (4 a 19). Nenhum paciente apresentou estenose de canal anal e 76% ficaram satisfeitos com a cirurgia com 90 dias de pós-operatório. Comparando-se os dados manométricos pré-operatórios e após 90 dias, nenhuma das variáveis avaliadas apresentou diferença com significância estatística. Não houve recidiva da doença hemorroidária com um ano de acompanhamento pós-operatório. Conclusão: a hemorroidopexia por grampeamento parcial não demonstrou impacto na fisiologia anorretal, apresentando baixos níveis de complicações e de dor pós-operatória, e sem recidivas após um ano de acompanhamento.

ABSTRACT Objective: to evaluate the impact of partial stapled hemorrhoidopexy on anorectal physiology, the complications related to this surgical technique, pain, postoperative bleeding and recurrence of hemorrhoidal disease one year after surgery. Methods: this is a prospective, descriptive study in consecutive patients with mixed or internal hemorrhoidal disease, the internal component being classified as grade III or IV, undergoing partial stapled hemorrhoidopexy. Results: we studied 17 patients, 82% of them with internal hemorrhoids grade III and 18% grade IV. The mean operative time was 09:09 minutes (07:03 to 12:13). The median pain in the immediate postoperative period evaluated by the numerical pain scale was one (0 to 7). The median time to return to work was nine days (4 to 19). No patient had anal stenosis and 76% were satisfied with the surgery 90 days postoperatively. When comparing the preoperative manometry data with that measured 90 days after surgery, none of the variables studied showed statistically significant difference. There was no recurrence of hemorrhoidal disease with one year of postoperative follow-up. Conclusion: partial stapled hemorrhoidopexy showed no impact on anorectal physiology, presenting low levels of complications and postoperative pain, without recurrence of hemorrhoidal disease in one year of follow-up.