Purinosome formation as a function of the cell cycle

The de novo purine biosynthetic pathway relies on six enzymes to catalyze the conversion of phosphoribosylpyrophosphate to inosine 5′-monophosphate. Under purine-depleted conditions, these enzymes form a multienzyme complex known as the purinosome. Previous studies have revealed the spatial organization and importance of the purinosome within mammalian cancer cells. In this study, time-lapse fluorescence microscopy was used to investigate the cell cycle dependency on purinosome formation in two cell models. Results in HeLa cells under purine-depleted conditions demonstrated a significantly higher number of cells with purinosomes in the G₁ phase, which was further confirmed by cell synchronization. HGPRT-deficient fibroblast cells also exhibited the greatest purinosome formation in the G₁ phase; however, elevated levels of purinosomes were also observed in the S and G₂/M phases. The observed variation in cell cycle-dependent purinosome formation between the two cell models tested can be attributed to differences in purine biosynthetic mechanisms. Our results demonstrate that purinosome formation is closely related to the cell cycle.Enzymes have been shown to form clusters in a cell to regulate metabolic processes (1–3). More recently, the concept of multienzyme complexes has been expanded to include mesoscale protein assemblies that appear to be substantially larger than a single protein (4). Depending on the metabolic or developmental state of the cells, these types of protein clusters range from transiently associated to very rigid and well defined (4). Examples of such enzyme clusters include the EF-Tu cytoskeleton, RNA degradosome, CTP synthase, carboxysomes, and nucleolus (5–9).In humans, purine nucleotides are synthesized by two different mechanisms. The first mechanism, de novo purine biosynthesis, converts phosphoribosylpyrophosphate (PRPP) to inosine 5′-phosphate (IMP) in 10 highly conserved steps catalyzed by six enzymes. These six enzymes include one trifunctional enzyme (TrifGART: GARS, GART, and AIRS domains), two bifunctional enzymes (PAICS: CAIRS and SAICARS domains; ATIC: AICART and IMPCH domains), and three monofunctional enzymes (PPAT, FGAMS, and ASL). ASL is also necessary for the conversion of IMP to AMP and may be classified as bifunctional (10). The second mechanism uses nucleotide salvage pathways to either phosphorylate a nucleoside (e.g., thymidine kinase) or add a purine base to ribose 5′-phosphate to regenerate the respective monophosphate. For example, hypoxanthine/guanine phosphoribosyl transferase (HGPRT) catalyzes the conversion of hypoxanthine to IMP and the conversion of guanine to guanosine 5′-phosphate (GMP). The de novo pathway is more energy-intensive, with the synthesis of 1 mole of IMP requiring 5 moles of ATP. Therefore, the salvage pathway is the preferred pathway for purine biosynthesis. Cells deficient in HGPRT, such as those that exhibit a Lesch–Nyhan disease (LND) phenotype, rely primarily on the de novo purine biosynthetic pathway to generate purine nucleotides (11–15).Recently, enzymes in the de novo purine biosynthetic pathway were shown to organize and reversibly assemble into punctate cellular bodies known as “purinosomes” under purine-depleted conditions (16). Further investigations into the organization of the purinosome showed that several of the enzymes form a core structure (PPAT, TrifGART, and FGAMS), whereas others appear to interact peripherally (PAICS, ASL, and ATIC) (17). The presence of this protein assembly and its putative function(s) clearly suggest that the spatial organization of pathway enzymes into purinosomes within a cell play an important role in meeting the cellular demand for purines (18). Although many studies have implied up-regulation of the de novo purine biosynthetic pathway during cell cycle progression (14, 19–21), here we used time-lapse microscopy to determine whether a correlation exists between the cell cycle stage and the number of cells with purinosomes or the cells’ purinosome content. Two different cell types, HeLa and LND cells, were used, with the latter deficient in purine salvage, to assess the effect of increased demand on the de novo pathway for purine biosynthesis and the attendant consequences for purinosome assembly.     

A case of abdominal bleeding after mitral valvuloplasty assisted by da Vinci robotic surgery

A 66-year-old female patient was admitted with abdominal bleeding as an unexpected complication of robotic surgery. Assessments included the patient's medical history, physical examination, laboratory data, and abdominal ultrasound scan results. In our case, laparotomy revealed an injury to the diaphragm and liver of the patient caused by the previous robotic surgery. In conclusion, although abdominal bleeding is a rare condition, it should be taken into consideration as a complication of robotic cardiac surgery.     

Exosome-delivered miR-221/222 exacerbates tumor liver metastasis by targeting SPINT1 in colorectal cancer

MicroRNAs (miRNAs) are involved in the progression of many cancers through largely unelucidated mechanisms. The results of our present study identified a gene cluster, miR-221/222, that is constitutively upregulated in serum exosome samples of patients with colorectal carcinoma (CRC) with liver metastasis (LM); this upregulation predicts a poor overall survival rate. Using an in vitro cell coculture model, we demonstrated that CRC exosomes harboring miR-221/222 activate liver hepatocyte growth factor (HGF) by suppressing SPINT1 expression. Importantly, miR-221/222 plays a key role in forming a favorable premetastatic niche (PMN) that leads to the aggressive nature of CRC, which was further shown through in vivo studies. Overall, our results show that exosomal miR-221/222 promotes CRC progression and may serve as a novel prognostic marker and therapeutic target for CRC with LM.     

Utility of feeding jejunostomy in patients with esophageal cancer undergoing esophagectomy with a high risk of anastomotic leakage

BackgroundFeeding jejunostomy is widely used for enteral nutrition (EN) after esophagectomy; however, its risks and benefits are still controversial. We aimed to evaluate the short-term and long-term outcomes of feeding jejunal tube (FJT) in patients undergoing esophagectomy for esophageal squamous cell carcinoma (ESCC) who were deemed high-risk for anastomotic leakage.MethodsWe retrospectively analyzed 716 patients who underwent esophagectomy with (FJT group, n=68) or without (control group, n=648) intraoperative placement of FJT. Propensity score matching (PSM) was used for the adjustment of confounding factors. Risk level for anastomotic leakage was determined for every patient after PSM.ResultsPatients in the FJT group were at higher risk of anastomotic leakage (14.9% vs. 11.3%), and had a statistically non-significant increase of postoperative complications [31.3% vs. 21.8%, odds ratio (OR) =1.139, 95% confidence interval (CI), 0.947–1.370, P=0.141] after PSM. Medical expenditure, length of postoperative hospital stay, and short-term mortality were similar between the FJT and control groups. Placement of FJT appeared to accelerate the recovery of anastomotic leakage (27.2 vs. 37.4 d, P=0.073). Patients in FJT group achieved comparable overall survival (OS) both before [hazard ratio (HR) =0.850, P=0.390] and after (HR =0.797, P=0.292) PSM.ConclusionsFJT showed acceptable safety profile along with potential benefits for ESCC patients with a high presumed risk of anastomotic leakage. While FJT does not impact OS, placement of FJT should be considered in esophagectomy patients and tailored to individual patients based on their leak-risk profile.     

γ-羟基丁酸对缺血/再灌注损伤后神经细胞凋亡及相关基因Bcl-2、Bax表达的影响

目的探讨γ-羟基丁酸(GHBA)对缺血/再灌注神经细胞凋亡及相关基因Bcl-2、Bax表达的影响及其可能的作用机制。方法采用大鼠局灶性缺血/再灌注损伤模型,大脑中动脉阻塞1h,分别于再灌注后1.5、3、6、12、24、48、72h,用原位末端缺口标记法(TUNEL)和免疫组织化学方法检测假手术组、生理盐水组和GHBA组海马区的凋亡细胞、Bcl-2、Bax的表达。结果与生理盐水组比较,GHBA组细胞凋亡显著减少(P<0.05),Bcl-2表达增多,Bax表达减少,且Bcl-2/Bax的比率上调(P<0.05)。结论GHBA可能通过增加Bcl-2表达、抑制Bax表达,抑制神经细胞凋亡,并可影响Bcl-2、Bax的平衡。     

Semantic processing of Chinese in left inferior prefrontal cortex studied with reversible words

This study utilized fast event-related fMRI with reversible words to examine the role of left inferior prefrontal cortex (PFC) in semantic processing of Chinese. As a special linguistic phenomenon in Chinese, a reversible word is a two-character word (AB) that, when read from right to left (BA), opposite to the normal left to right reading direction, is also a real word. The two words, AB and BA, can have very different meanings. Fourteen native Chinese saw a reversible word (BA) and were asked to read it backward silently to obtain the meaning of AB, defined as the target meaning. They then saw two test words and decided which of the two was semantically related to the target meaning. Activity in a subregion of BA47 was found to be modulated by the extent to which irrelevant semantic activation of the distractor word BA interfered with semantic retrieval of the target word AB. This finding demonstrated the involvement of the left inferior PFC in the control processes of semantic retrieval in Chinese. In addition, comparing conditions using reversible with that using nonreversible words, we found evidence suggesting a semantic/phonological functional subdivision in left inferior PFC, consistent with that in English.