Dendritic cells derived from murine colonic mucosa have unique functional and phenotypic characteristics

Dendritic cells (DCs) residing in different tissues and exposed to different organisms are likely to have different reactivities to their surrounding environment. Many studies use in vitro generated DCs to examine functions of these cells, but such cells may not truly reflect the nature of DCs and their in situ activities in vivo. We have used magnetic label-based technique to isolate colonic DCs to conduct derailed characterization of these cells. Colonic DCs comprise mainly CD11b+ DCs with few CD8alpha+ DCs or plasmacytoid DCs. Functionally, isolated colonic DCs are able to endocytose and process proteins, undergo maturation, and stimulate T cells to proliferate. Importantly, expression of TLRs by colonic DCs is significantly lower than that of their spleen counterparts; however, they appear to be as, or more, responsive to stimulation by oligodeoxynucleotides containing CpG motif based on their cytokine production. We speculate that colonic DCs have unique reactivities differing from DCs residing in other lymphoid tissues and are adapted for the unique microenvironment of the colonic mucosa and that these cells react uniquely to their environment.     

PECAM-1 shedding during apoptosis generates a membrane-anchored truncated molecule with unique signaling characteristics.

Shedding of cell surface molecules, including growth factor receptors, provides a mechanism by which cells regulate signal transduction events. Here we show that platelet-endothelial cell adhesion molecule (PECAM)-1 is shed from the endothelial cell surface during apoptosis and accumulates in the culture medium as a approximately 100 kDa soluble protein. The cleavage mediating the shedding is matrix metalloproteinase (MMP) dependent, as GM6001, a broad-spectrum MMP inhibitor, inhibits PECAM-1 accumulation in the culture medium in a dose-responsive manner. In addition to the 100 kDa soluble fragment, PECAM-1 cleavage generates the formation of a truncated (Tr.) approximately 28 kDa molecule, composed of the transmembrane and the cytoplasmic PECAM-1 domains. Transfections of the full-length (Fl) and the Tr. PECAM-1 gene constructs into endothelial and nonendothelial cells were performed. We found 1) significantly more gamma-catenin and SHP-2 bound to the truncated than to the full-length PECAM-1; 2) stable expression of the truncated PECAM-1 in SW480 colon carcinoma cells resulted in a dramatic decrease in cell proliferation, whereas expression of comparable levels of the full-length PECAM-1 had no effect; 3) the decrease observed in cell proliferation is due, in part, to an increase in programmed cell death (apoptosis) and correlated with continuous caspase 8 cleavage and p38/JNK phosphorylation. These results support the intimate involvement of PECAM-1 in signal transduction cascades and also suggest that caspase substrates (e.g., PECAM-1) may possess distinct and unique functions on cleavage.     

Predictors of mortality of patients newly diagnosed with clinical type 2 diabetes: a 5-year follow up study

Background

At diabetes diagnosis major decisions about life-style changes and treatments are made based on characteristics measured shortly after diagnosis. The predictive value for mortality of these early characteristics is widely unknown. We examined the predictive value of patient characteristics measured shortly after diabetes diagnosis for 5-year all-cause and cardiovascular mortality with special reference to self-rated general health.

Methods

Data were from a population-based sample of 1,323 persons newly diagnosed with clinical diabetes and aged 40 years or over. Possible predictors of mortality were investigated in Cox regression models.

Results

Multivariately patients who rated their health less than excellent experienced increased all-cause and cardiovascular mortality. These end-points also increased with sedentary life-style, relatively young age at diagnosis and presence of cardiovascular disease (CVD) at diagnosis. Further predictors of all-cause mortality were male sex, low body mass index and cancer, while cardiovascular mortality increased with urinary albumin concentration.

Conclusions

We found that patients who rated their health as less than excellent had increased 5-year mortality, similar to that of patients with prevalent CVD, even when biochemical, clinical and life-style variables were controlled for. This finding could motivate doctors to discuss perceptions of health with newly diagnosed diabetic patients and be attentive to patients with suboptimal health ratings. Our findings also confirm that life-style changes and optimizing treatment are particularly relevant for relatively young and inactive patients and those who already have CVD or (micro)albuminuria at the time of diabetes diagnosis.     

甘精胰岛素联用瑞格列奈治疗初诊2型糖尿病的疗效研究

目的:通过甘精胰岛素联用瑞格列奈与预混人工合成胰岛素(诺和灵30R)治疗初诊2型糖尿病患者的比较,探讨其疗效与安全性.方法:将初诊2型糖尿病患者随机分为甘精胰岛素+瑞格列奈组(A组)和诺和灵30R组(B组),根据血糖情况调整用药剂量.治疗12周后,比较两组的空腹血糖、餐后2小时血糖、糖化血红蛋白(HbA1c)、体重指数(BMI)和低血糖发生率.结果:A组低血糖事件明显少于B组,在餐后2小时血糖方面也优于B组,差异有统计学意义(P<0.05);在空腹血糖、HbA1c和BMI方面差异无统计学意义(p0.05).结论:甘精胰岛素与瑞格列奈联用对于初诊2型糖尿病患者,其血糖控制满意,餐后血糖更加平稳,低血糖发生率低,是一种针对初诊2型糖尿病患者安全、有效、方便的治疗方案.     

New human liver alcohol dehydrogenase forms with unique kinetic characteristics

All adult and infant human liver homogenates studied thus far show two previously unreported forms of alcohol dehydrogenase on starch gel electrophoresis. Under the conditions employed, these forms migrate toward the anode and readily stain for pentanol but virtually not for ethanol oxidizing activity. In contrast, all human ADH isoenzymes identified previously are cathodic and react equally well with either substrate. These new ADH forms have been separated from the other known ones by DEAE-cellulose chromatography and are then purified on Agarose-hexane-AMP. Although the physical characteristics of the new anodic ADH forms are similar to those of the known human ADH isoenzymes, the former are not inhibited by 12 mM 4-methyl pyrazole, oxidize ethanol very poorly and appear to prefer longer chain alcohols as substrates.     

Barrett esophagus diagnosed after appendectomy in a patient with diabetes mellitus]

A pathogenetic outline of oesophageal inflammatory lesions with special emphasis on Barrett's oesophagitis has been presented. As recorded in a history of a 65-year old patient at first the stenocardia-mimicking pains occurred a few days after appendectomy. Gastrobulboscopy performed (after coronary insufficiency had been excluded) revealed extensive inflammatory and erosive lesions of the oesophagus and multiple ulcerations of the duodenum. According to the several authors the rapid manifestation of the inflammatory lesions of the oesophagus may be related to the intubation necessary for laparotomy. It may refer particularly to diabetic patients. Control endoscopies showed that a six-week treatment resulted in a total disappearance of mucosal changes. It might be thoughtful to take into consideration a possibility of oesophageal inflammation in clinical differentiation of stenocardia mimicking pains in diabetics who underwent the intubation.     

Palm tree peroxidase-based biosensor with unique characteristics for hydrogen peroxide monitoring

Three amperometric enzyme electrodes have been constructed by adsorbing anionic royal palm tree peroxidase (RPTP), anionic sweet potato peroxidase (SPP), or cationic horseradish peroxidase (HRP-C) on spectroscopic graphite electrodes. The resulting H(2)O(2)-sensitive biosensors were characterized both in a flow injection system and in batch mode to evaluate their main bioelectrochemical parameters, such as pH dependency, I(max), K(M)(app), detection limit, linear range, operational and storage stability. The obtained results showed a distinctly different behavior for the plant peroxidase electrodes, demonstrating uniquely superior characteristics of the RPTP-based sensors. The broader linear range observed for the RPTP-based biosensor is explained by a high stability of this enzyme in presence of H(2)O(2). The higher storage and operational stability of RPTP-based biosensor as well as its capability to measure hydrogen peroxide under acidic conditions connect with an extremely high thermal and pH-stability of RPTP.     

Annexin V staining during reperfusion detects cardiomyocytes with unique properties

With the use of markers of sarcolemmal membrane permeability, cardiomyocyte models of ischemic injury have primarily addressed necrotic death during ischemia. In the present study, we used annexin V-propidium iodide staining to examine apoptosis and necrosis after simulated ischemia and simulated reperfusion in rat ventricular myocytes. Annexin V binds phosphatidylserine, a phosphoaminolipid thought to be externalized during apoptosis or programmed cell death. Propidium iodide is a marker of cell necrosis. Under baseline conditions, <1% of cardiomyocytes stained positive for annexin V. After 20 or 60 min of simulated ischemia, there was no increase in annexin V staining, although 60-min simulated ischemia resulted in significant propidium iodide staining. Twenty minutes of simulated ischemia, followed by 20 or 60 min of simulated reperfusion, resulted in 8-10% of myocytes staining positive for annexin V. Annexin V-positive cells retained both rod-shaped morphology and contractile function but exhibited the decreased cell width indicative of cell shrinkage. Baseline mitochondrial free Ca2+ (111 +/- 14 nM) was elevated in reperfused annexin V-negative cells (214 +/- 22 nM), and further elevated in annexin V-positive myocytes (382 +/- 9 nM). After 60 min of simulated reperfusion, caspase-3-like activity was observed in approximately 3% of myocytes, which had a rounded appearance and membrane blebs. These results suggest that the use of annexin V after simulated ischemia-reperfusion uncovers a population of cardiomyocytes whose characteristics appear to be consistent with cells undergoing apoptosis.     

GABAA receptor subtypes immunopurified from rat brain with alpha subunit-specific antibodies have unique pharmacological properties.

The unique cytoplasmic loop regions of the alpha 1, alpha 2, alpha 3, and alpha 5 subunits of the GABAA receptor were expressed in bacterial and used to produce subunit-specific polyclonal antisera. Antibodies immobilized on protein A-Sepharose were used to isolate naturally occurring alpha-specific populations of GABAA receptors from rat brain that retained the ability to bind [3H]muscimol, [3H]flunitrazepam, [3H]Ro15-1788, and [125I]iodo-clonazepam with high affinity. Pharmacological characterization of these subtypes revealed marked differences between the isolated receptor populations and was generally in agreement with the reported pharmacological profiles of GABAA receptors in cells transiently transfected with alpha 1 beta 1 gamma 2, alpha 2 beta 1 gamma 2, alpha 3 beta 1 gamma 2, and alpha 5 beta 1 gamma 2 combinations of subunits. Additional subtypes were also identified that bind [3H]muscimol but do not bind benzodiazepines with high affinity. The majority of GABAA receptor oligomers contains only a single type of alpha subunit, and we conclude that alpha 1, alpha 2, alpha 3, and alpha 5 subunits exist in vivo in combination with the beta subunit and gamma 2 subunit.     

Children with type 2 diabetes: the risks of complications

Type 2 diabetes is an increasing problem in children. Two decades ago it had been described only in selected groups, e.g. the Pima Indians. Childhood type 2 diabetes appears to be similar to the metabolic syndrome in adults and is characterized by obesity, hyperglycaemia and insulin resistance. It can present a diagnostic challenge in children, as they can present with diabetic ketoacidosis; the measurement of autoantibodies and C-peptide levels may be helpful. The logarithmic association between the risk of complications with increasing glycaemia which has been established for adults with type 2 diabetes is likely to hold true for children but the conclusions of trials in adults must be extrapolated with caution. Little is known about the onset and progression of macrovascular disease in affected children but it is almost certain that they will develop an excess of premature cardiovascular disease. However, the importance of reducing glycaemia in younger adults with diabetes, in order to minimize the incidence of microvascular complications, has been unequivocally demonstrated in the Diabetes Control and Complications Trial (DCCT). Diet and exercise have a major role to play in the treatment and prevention of type 2 diabetes in children as well as adults - the escalation of type 2 diabetes throughout the developed world is a major public health problem. Extrapolating data from adults, metformin appears to be the logical first-line treatment in children with type 2 diabetes; sulphonylureas are also used but neither of these agents have been evaluated in trials in children and are not licensed for such use. With regard to other newer agents, it seems wise to use well-established drugs with a long track record and for which the long-term safety data are available.     

A homodimeric laccase with unique characteristics from the yellow mushroom Cantharellus cibarius

The aim of the present study was to isolate a laccase from fruiting bodies of the yellow mushroom Cantharellus cibarius. The fruiting body extract was subjected to a purification protocol that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel and Con A-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The laccase was unadsorbed on DEAE-cellulose and Affi-gel blue gel and adsorbed on Con A-Sepharose. The laccase was composed of two identical subunits each with a molecular mass of 46 kDa. The laccase exhibited a temperature-dependent rise in activity over the temperature range 20-50 degrees C. When the temperature was raised above 60 degrees C there was a fall in enzyme activity. The enzyme manifested maximal activity at pH 4. At and above pH 6 there was a dramatic reduction in activity. The unique features of this fruiting body laccase compared with previously reported mycelial laccases include homodimeric nature, a distinctive N-terminal sequence, a higher optimal pH, and adsorption on only ConA-Sepharose among the various chromatographic media tested.     

Children with idiopathic hemihypertrophy and beckwith-wiedemann syndrome have different constitutional epigenotypes associated with wilms tumor

Idiopathic hemihypertrophy (IH) is a congenital overgrowth syndrome associated with an increased risk of embryonal cancers in childhood. A related developmental disorder is Beckwith-Wiedemann syndrome (BWS), which increases risk for embryonal cancers, including Wilms tumor. Constitutional epigenetic alterations associated with BWS have been well characterized and include epigenetic alterations of imprinted genes on 11p15. The frequency of hypermethylation of H19 in children with IH and Wilms tumor, 20% (3/15), was significantly lower than the frequency in children with BWS and Wilms tumor, 79% (11/14; P = .0028). These results indicate that children with IH and Wilms tumor have different constitutional epigenotypes from those of children with BWS and Wilms tumor.