Application of the SpCas9 inhibitor BRD0539 for CRISPR/Cas9-based genetic tools in .

Although the clustered regularly interspaced short palindromic repeat (CRISPR)-Cas system has been extensively developed since its discovery for eukaryotic and prokaryotic genome editing and other genetic manipulations, there are still areas warranting improvement, especially regarding bacteria. In this study, BRD0539, a small-molecule inhibitor of Cas9 (SpCas9), was used to suppress the activity of the nuclease during genetic modification of , as well as to regulate CRISPR interference (CRISPRi). First, we developed and validated a CRISPR-SpCas9 system targeting the gene of .

Preoperative difficulty factors in delayed laparoscopic cholecystectomy: Tokyo Guidelines 2018 surgical difficulty score analysis.

Introduction: Tokyo Guidelines 2018 (TG18) recommend early laparoscopic cholecystectomy (LC) for low-risk acute cholecystitis (AC); however, some patients undergo delayed LC (DLC) after conservative treatment. DLC, influenced by chronic inflammation, is a difficult procedure. Previous studies on LC difficulty lacked objective measures.

[Two Cases of Stage Ⅳ Colorectal Cancer with Long-Term Survival following Single-Agent Chemotherapy with Capecitabine-A Case Reports].

We experienced 2 cases of Stage Ⅳ colorectal cancer obtained long-term survival by chemotherapy with only capecitabine. Case 1: Seventy-one-years-old male was performed open sigmoid colectomy, D2 dissection for sigmoid colon cancer. Pathological diagnosis was pT4aN2aM0, pStage Ⅲc.

Disulfide bond formation in microtubule-associated tau protein promotes tau accumulation and toxicity in vivo.

Accumulation of microtubule-associated tau protein is thought to cause neuron loss in a group of neurodegenerative diseases called tauopathies. In diseased brains, tau molecules adopt pathological structures that propagate into insoluble forms with disease-specific patterns. Several types of posttranslational modifications in tau are known to modulate its aggregation propensity in vitro, but their influence on tau accumulation and toxicity at the whole-organism level has not been fully elucidated.

[A Case of Histological Complete Remission after Chemotherapy for Paraaortic Lymph Node Recurrence after Colon Cancer Surgery].

In September 2013, a 50-year-old woman presented to our hospital with right abdominal pain as the main complaint. Careful examinations led to the diagnosis of ascending colon cancer. In October 2013, resection of the right half colon and removal of the D3 lymph nodes were performed.

[Two Cases of Locally Advanced Rectal Cancer and Lower Rectal Cancer Resected Successfully That Enabled Anus Preservation after Preoperative Chemotherapy].

We reported 2 cases of colorectal cancer receiving neoadjuvant chemotherapy(NAC)with the aim of curative resection or anal preservation. Case 1: A 50-year-old man was diagnosed with locally advanced rectal cancer with sacral invasion. Because of the sacral invasion, we performed preoperative chemotherapy.

S6K/p70S6K1 protects against tau-mediated neurodegeneration by decreasing the level of tau phosphorylated at Ser262 in a Drosophila model of tauopathy.

Abnormal accumulation of the microtubule-associated protein tau is thought to cause neuronal cell death in a group of age-associated neurodegenerative disorders. Tau is phosphorylated at multiple sites in diseased brains, and phosphorylation of tau at Ser262 initiates tau accumulation and toxicity. In this study, we sought to identify novel factors that affect the metabolism and toxicity of tau phosphorylated at Ser262 (pSer262-tau).

Postherpetic pseudohernia: delayed onset of paresis of abdominal muscles due to herpes zoster causing an ipsilateral abdominal bulge.

Postherpetic pseudohernia causes an abdominal bulge as well as an abdominal wall herniation. This disease is one of the neurological complications of herpes zoster and essentially consists of paresis of ipsilateral abdominal muscles. Postherpetic pseudohernia may be mistaken for abdominal wall herniation because it is not well known.

[Two Cases of Rectal Cancer Resected Curatively after Chemotherapy with CapeOX plus Bmab].

We report 2 cases of locally far-advanced rectosigmoid cancer that were initially unresectable, but were successfully excised after treatment with CapeOX plus Bmab chemotherapy(capecitabine, L-OHP, and bevacizumab). Case 1: A 72-year-old man who complained of severe constipation initially received sigmoid colostomy because of far-advanced rectosigmoid cancer. After 4 courses of CapeOX plus Bmab chemotherapy administration, the size of the primary tumor remarkably decreased and curative resection could be performed.

Assay methods for H2S biogenesis and catabolism enzymes.

H2S is produced from sulfur-containing amino acids, cysteine and homocysteine, or a catabolite, 3-mercaptopyruvate, by three known enzymes: cystathionine β-synthase, γ-cystathionase, and 3-mercaptopyruvate sulfurtransferase. Of these, the first two enzymes reside in the cytoplasm and comprise the transsulfuration pathway, while the third enzyme is found both in the cytoplasm and in the mitochondrion. The following mitochondrial enzymes oxidize H2S: sulfide quinone oxidoreductase, sulfur dioxygenase, rhodanese, and sulfite oxidase.

Structure and kinetic analysis of H2S production by human mercaptopyruvate sulfurtransferase.

Mercaptopyruvate sulfurtransferase (MST) is a source of endogenous H2S, a gaseous signaling molecule implicated in a wide range of physiological processes. The contribution of MST versus the other two H2S generators, cystathionine β-synthase and γ-cystathionase, has been difficult to evaluate because many studies on MST have been conducted at high pH and have used varied reaction conditions. In this study, we have expressed, purified, and crystallized human MST in the presence of the substrate 3-mercaptopyruvate (3-MP).

Minimal incision transinguinal repair for incarcerated obturator hernia.

Background: Patients with incarcerated obturator hernia are usually elderly, frail, and physically inactive women with serious comorbidities. Although a laparotomy is standard surgical intervention for emergency incarcerated or strangulated obturator hernia, it is invasive particularly for these high-risk patients. The aim of this study is to show the feasibility of minimum open inguinal approach to reduce surgical risk for preoperatively diagnosed incarcerated obturator hernia.

A case report of undifferentiated carcinoma with osteoclast-like giant cells of the pancreas and literature review.

Osteoclast-like giant cell tumors rarely arise in the pancreas. Here we report the case of a 78-year-old woman who was diagnosed with a well-defined 3 cm multilocular mass in the pancreatic body by the use of ultrasonography, computed tomography and magnetic resonance imaging. The rim and the septa of the tumor were well enhanced.

Relative contributions of cystathionine beta-synthase and gamma-cystathionase to H2S biogenesis via alternative trans-sulfuration reactions.

In mammals, the two enzymes in the trans-sulfuration pathway, cystathionine beta-synthase (CBS) and cystathionine gamma-lyase (CSE), are believed to be chiefly responsible for hydrogen sulfide (H2S) biogenesis. In this study, we report a detailed kinetic analysis of the human and yeast CBS-catalyzed reactions that result in H2S generation. CBS from both organisms shows a marked preference for H2S generation by beta-replacement of cysteine by homocysteine.

H2S biogenesis by human cystathionine gamma-lyase leads to the novel sulfur metabolites lanthionine and homolanthionine and is responsive to the grade of hyperhomocysteinemia.

Although there is a growing recognition of the significance of hydrogen sulfide (H(2)S) as a biological signaling molecule involved in vascular and nervous system functions, its biogenesis and regulation are poorly understood. It is widely assumed that desulfhydration of cysteine is the major source of H(2)S in mammals and is catalyzed by the transsulfuration pathway enzymes, cystathionine beta-synthase and cystathionine gamma-lyase (CSE). In this study, we demonstrate that the profligacy of human CSE results in a variety of reactions that generate H(2)S from cysteine and homocysteine.

A dithio-coupled kinase and ATPase assay.

Kinases and ATPases produce adenosine diphosphate (ADP) as a common product, so an assay that detects ADP would provide a universal means for activity-based screening of enzymes in these families. Because it is known that most kinases accept ATPbetaS (sulfur on the beta-phosphorous) as a substrate in place of adenosine triphosphate (ATP), the authors have developed a continuous assay using this substrate, with detection of the ADPbetaS product using dithio reagents. Such an assay is possible because dithio groups react selectively with ADPbetaS and not with ATPbetaS.

Fluorescence-based detection of thiols in vitro and in vivo using dithiol probes.

Thiols play a central role in maintaining biological homeostasis. Their levels can change dramatically in response to oxidative stress associated with toxic insults, bacterial infection, and disease. Therefore, a reagent that can monitor thiol levels both in vitro and in vivo would be useful for assays and as a biomarker.

Simple way to improve accuracy in diagnosis of quadrant inflammatory disease: how to avoid adverse laparotomy by using plain CT.

Background/aims: It has been reported that contrast computed tomography (CT) provides a rapid and accurate diagnosis of acute appendicitis, and could potentially prevent adverse appendectomy. In the present study, we evaluated the diagnostic value of precontrast CT combined with that of serum C reactive protein (CRP) concentration for right lower quadrant inflammatory diseases.

Methodology: One hundred consecutive patients who had rebound tenderness and muscular rigidity or guarding on right lower quadrant underwent an emergency abdominal CT without contrast medium and laboratory tests.

Clinical effects of long-term use of neutralized dialysate for continuous ambulatory peritoneal dialysis.

The long-term effects of neutralized dialysate used in continuous ambulatory peritoneal dialysis (CAPD) were evaluated in 8 well-controlled patients. Twelve milliliters of 8.4% sodium bicarbonate was added to Dianeal PD-1 immediately before every administration.