Ubiquitin hybrid protein gene expression during human colon cancer progression

Ubiquitin is involved in cell-cycle control and DNA replication through a specific proteolytic pathway. Our previous studies demonstrated selected higher expression of a gene encoding ubiquitin-ribosomal protein S27a in poorly differentiated colon carcinoma cell lines. In this study, we evaluated this ubiquitin hybrid protein gene expression in surgical specimens of colon cancers. Northern blot analysis showed that ubiquitin hybrid protein messenger RNA was overexpressed in primary colon cancers compared with adjacent normal colon mucosae in 17 of 20 patients. Dot blot analysis of RNA of 27 tumor samples revealed significantly greater expression in higher Dukes' stage primary colon tumors and liver metastases. These data imply that protein translation machinery is highly activated during progression and metastasis of colon tumors, and that ubiquitin hybrid protein may be useful as a marker of biological aggressiveness.     

Bouveret meets Boerhaave

Rarely, biliary-enteric fistula can result in duodenal obstruction or Bouveret's syndrome. Boerhaave's syndrome is a distal esophageal rupture in the setting of severe emesis. This case is the first reported successful management of these clinical scenarios occurring simultaneously and highlights important features in presentation, diagnosis, and surgical treatment.     

Exocrine meets endocrine: pancreatic stone protein and regenerating protein--two sides of the same coin

BACKGROUND: Regenerating protein (reg) and pancreatic stone protein (PSP) have been discovered independently in the fields of diabetes and pancreatitis. MATERIALS AND METHODS: These proteins are identical; however, because of the gap between the endocrine and exocrine field, there was never a consensus and the nomenclature has not been rectified. Since the time of the initial discovery, more isoforms have been unified. Historically, PSP was discovered long before reg, yet, in many areas outside of the pancreatitis research field, reg is being used. RESULTS: For PSP/reg, a role in proliferation and regeneration of islet cells has been postulated. A hitherto insufficiently understood phenomenon is the massive up-regulation of PSP/reg in pancreatic tissue and juice under conditions of stress. Similarly, PAP (pancreatitis-associated protein)/reg III has been attributed various functional roles. Structurally, the ability to form fibrils after tryptic cleavage is a striking common features of both proteins. However, this biochemical transformation is in itself not enough to gain functional insight. Thus, physiological and genetic approaches are required to further characterize the role of these proteins in the pancreas. Recently, more evidence has been presented in support of the theory that PSP/reg plays a key role in islet neogenesis/regeneration. CONCLUSIONS: In this review we discuss the debate on the localization and functional roles of PSP/reg and PAP/regIII. Therefore, we have summarized hypotheses and experimental results supporting such hypotheses.     

Ubiquitin reduces fluid shifts after traumatic brain injury

BACKGROUND: Ubiquitin has well-described intracellular properties. Recent data also suggest pleiotropic effects of extracellular ubiquitin, including induction of apoptosis, regulation of immune functions, and therapeutic potential during fluid resuscitation from severe trauma. However, the actions of exogenous ubiquitin after traumatic brain injury (TBI) are unknown. METHODS: Series 1: Thirty-five minutes after TBI and hemorrhage, 1.5 mg ubiquitin/kg (n = 5) or albumin (n = 5) intravenous was followed by fluid resuscitation to maintain mean arterial and cerebral perfusion pressure. Series 2: Ubiquitin (n = 5) or vehicle (n = 6) was administered after TBI only. Ubiquitin was measured with enzyme-linked immunosorbent assay in serum, urine (series 1), and cerebrospinal fluid (series 2) for 300 minutes. RESULTS: Series 1: After intravenous bolus, serum ubiquitin peaked at t = 45 minutes with a half-life of 54 minutes. Recovery in urine was 10%. With albumin versus ubiquitin, 85% more resuscitation fluid was required to stabilize systemic and cerebral hemodynamics (P < .05 for t = 150 to 300 minutes), but hematocrit was similar. With albumin there were progressive increases in intracranial pressure, peak inspiratory pressure, and decreases in oxygenation. All were significantly attenuated by ubiquitin (all P < .05 vs albumin). Series 2: Intravenous ubiquitin altered cerebrospinal fluid ubiquitin with an increased time to peak (t = 88 +/- 13 min vs 45 +/- 7 min, P < .05) and area under the concentration-time curve (82 +/- 22 vs 23 +/- 11 microg/min(1)/mL(-1), P < .05). CONCLUSIONS: After TBI, intravenous ubiquitin crossed the blood-brain barrier and significantly reduced third spacing of fluid into the brain and lung during resuscitation.     

Ubiquitin and malignant transformation of oral mucosa.

BACKGROUND: Ubiquitinylation is a cyclical process operating in all cells to target specific proteins (eg, p53) for degradation. Abnormal accumulations of ubiquitinylated proteins have been identified in colorectal carcinoma. This study investigated abnormal ubiquitinylated proteins and changes in ubiquitinylation patterns associated with malignant transformation of oral tissue. METHODS: Twenty-three fresh specimens of normal, premalignant, oral squamous cell carcinoma (SCC), and its adjacent normal mucosa were collected for immunohistochemistry with anti-ubiquitin and anti-UbcH7 (a human ubiquitin-conjugating enzyme) antibodies using light and electron microscopy and protein analysis by Western blot. RESULTS: Ubiquitin and UbcH7 were observed in both nucleus and cytoplasm of normal, "premalignant" and malignant tissues of oral mucosa using immunoelectron microscopy. However there was no change in their distribution pattern. CONCLUSIONS: The lack of demonstrable difference in distribution pattern of ubiquitin enzymes indicates that at present, ubiquitin cannot be used as a diagnostic marker for malignant transformation of oral SCC.     

Hirschsprung's disease: healthcare meets the needs

Objective

The objective of this study was to examine the types of healthcare services used by children, adolescents, and adults with Hirschsprung Disease (HD) in relation to the severity of the initial defect (mild, severe), whether additional care was needed, the provision of information, transfer to adult care, and satisfaction with the care provided.

Methods

Three hundred twenty (71%) HD patients, ages 6 to 54, completed a questionnaire that assessed the use of healthcare services, the need for more healthcare, the provision of information, the transfer to adult care and satisfaction with the provided care.

Results

In 6 months, 45% of the children, 14% of the adolescents, and 15% of the adults consulted a medical specialist. Compared with patients with a mild form of HD in the age range of 6 to 16 years, only the more severely afflicted adult patients visited medical professionals more often (10% vs 29%) (P < .05). Of the children, the adolescents, and the adults 23%, 8%, and 6% respectively consulted a nonmedical professional. Less than 15% of all patients whould have liked more treatment. In 6 months 51% of the children, 24% of the adolescents, and 21% of the adults received treatment information, of which respectively 14, 8, and 20% wished they had received more information. Three (12%) patients who needed adult care encountered problems with the transfer. Almost all patients were satisfied with the care provided.

Conclusions

There is good access to medical healthcare services, especially for children. The only lacuna in the healthcare system we revealed was a lack of information, particularly for adult patients. Most parents and patients reported to be very satisfied with the provided care.     

Ubiquitin C-terminal hydrolase L1 after out-of-hospital cardiac arrest

Background

We studied the prognostic ability of serum ubiquitin C-terminal hydrolase L1 (UCH-L1) after out-of-hospital cardiac arrest (OHCA), compared to that of neuron-specific enolase (NSE).

Methods

In this post-hoc analysis of the FINNRESUSCI study, we measured serum concentrations of UCH-L1 in 249 OHCA patients treated in 21 Finnish intensive care units in 2010–2011. We evaluated the ability of UCH-L1 to predict unfavourable outcome at 12 months (defined as cerebral performance category 3–5) by assessing the area under the receiver operating characteristic curve (AUROC), in comparison with NSE.

Results

The concentrations of UCH-L1 were higher in patients with unfavourable outcome than for those with favourable outcome: median concentration 10.8 ng/mL (interquartile range, 7.5–18.5 ng/mL) versus 7.8 ng/mL (5.9–11.8 ng/mL) at 24 h (p < .001), and 16.2 ng/mL (12.2–27.7 ng/mL) versus 11.5 ng/mL (9.0–17.2 ng/mL) (p < .001) at 48 h after OHCA. For UCH-L1 as a 12-month outcome predictor, the AUROC was 0.66 (95% confidence interval, 0.60–0.73) at 24 h and 0.66 (0.59–0.74) at 48 h. For NSE, the AUROC was 0.66 (0.59–0.73) at 24 h and 0.72 (0.65–0.80) at 48 h. The prognostic ability of UCH-L1 was not different from that of NSE at 24 h (p = .82) and at 48 h (p = .23).

Conclusion

Concentrations of UCH-L1 in serum were higher in patients with unfavourable outcome than in those with favourable outcome. However, the ability of UCH-L1 to predict unfavourable outcome after OHCA was only moderate and not superior to that of NSE.     

泛素在大鼠慢性同种移植肾病中的表达及作用

目的:观察泛素在慢性移植肾病(CAN)大鼠肾组织中的表达及其与肾功能改变、病理学改变及肾组织正常活化T细胞表达及分泌趋化因子(RANTES)之间的关系,探讨泛素在CAN中的作用.方法:实验组采用近交系大鼠的同种异基因动物间的左肾原位移植,雄性F344大鼠40只为供体,雄性LEW大鼠40只为受体,移植手术后第10 d行右肾切除术,对照组采用单肾切除术的雄性F344和LEW大鼠各25只.分别于4周、8周、12周、16周及24周处死大鼠,做肾功能、移植肾组织学检测,并应用免疫组织化学与免疫印迹方法检测肾组织中泛素的表达,应用免疫组织化学方法检测肾组织RANTES的表达.结果:移植组大鼠尿蛋白定量、血清肌酐水平于移植后16周时显著增高,24周时更为显著.肾脏病理显示:移植后12周之前,主要表现为肾间质单个核细胞浸润,血管平滑肌细胞增殖及血管内膜的断裂;移植后16周时可观察到肾小球硬化、肾间质纤维化及血管硬化等慢性病理改变.免疫组织化学及Western-blot结果显示泛素在CAN病程中的表达呈逐渐升高后又降低的过程,移植12周为其表达高峰,后逐渐降低,24周时降至最低.免疫组织化学结果显示RANTES表达部位,表达时相与泛素表达是一致的.相关分析显示,肾移植大鼠泛素表达水平与24 h尿蛋白定量、血清肌酐水平在CAN早期呈正相关,在CAN晚期呈负相关.与肾间质纤维化程度呈负相关.移植12周组泛素表达水平与RANTES呈显著正相关.结论:泛素在CAN早晚期病变中起连接作用,泛素对上调增殖起基本作用,并可能通过对RANTES的调节,参与了CAN的全过程.