Heat shock suppresses human NK cell cytotoxicity via regulation of perforin.

Human natural killer (NK) cell, which is an important lymphocyte for immune surveillance, is highly sensitive to heat, but the nature of its response to and its mechanistic regulation by heat remain unclear. Here we determined the effect of in vitro heat shock and in vivo hyperthermia on human NK cell cytotoxicity. Human peripheral blood mononuclear cells (PBMC) obtained from healthy volunteers were subjected to heat shock in vitro (42 degrees C, 1 h). PBMC from cancer patients receiving intentional hyperthermia (42 degrees C, 1 h) for cancer therapy were also obtained. NK cytolytic activity was determined in these samples. NK cell cytotoxicity was down-regulated by heat shock in vitro at 5 h, but at 24 h after heat shock, the NK cytotoxicity was comparable to that with its respective control. Furthermore, we observed that the mRNA and protein expression levels of perforin, which is the cytolytic granule of NK cells, were regulated by heat shock in a similar manner as NK cytotoxicity at 5 h and at 24 h after heat shock. Heat regulation involved the perforin protein in CD56(dim) but not in CD56(bright) NK cell subset. Heat shock neither induced cell death nor altered the expression of some NK activating receptors and adhesion molecules. Moreover, whole-body hyperthermia at 42 degrees C for 1 h of cancer patients also suppressed the cytotoxicity of NK cells but recovered to basal level 1 week after hyperthermia. Heat shock in vitro and in vivo temporarily represses the cytotoxicity of human NK cells.     

Coenzyme specificity in fungal 17beta-hydroxysteroid dehydrogenase

The 17beta-hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus is an NADP(H)-dependent member of the short-chain dehydrogenase/reductase superfamily (SDR) that belongs to the cP1 classical subfamily. Here, we have created several mutants by site-directed mutagenesis, and through these we have studied the amino acid residues that are responsible for coenzyme binding and specificity. The Thr202Val and Thr202Ile mutants were inactive, thus confirming the importance of Thr202 for the appropriate orientation of the coenzyme that enables the hydride transfer. The Ala50Arg and Asn51Arg mutants had increased rates of NADPH dissociation, and thus an enhanced substrate oxidation with NADP+, while the Asn51Arg mutant also showed an increased rate of NADP+ dissociation, and thus an enhanced substrate reduction with NADPH. Addition of a negatively-charged amino acid residue at the first position after the second beta-strand (Tyr49Asp) affected the coenzyme specificity and turned the enzyme into an NAD+-dependent oxidase resembling the cD1d subfamily members.     

On-Pump Beating Heart Surgery Offers an Alternative for Unstable Patients Undergoing Coronary Artery Bypass Grafting

Abstract Background: Cardiac surgery has expanded the available approaches to aortocoronary artery bypass grafting to include approaches from minimally invasive surgery to full sternotomy. The heart can be arrested, left beating, or assisted with a right ventricular assist device or cardiopulmonary bypass pump. We have examined the 4 surgical modes that we use routinely in our large multisurgeon practice to determine our selection biases and the outcomes of the different techniques. Methods: Of the 4733 coronary artery bypass grafting (CABG) patients we studied from January 2000 through December 2002, 2332 (49.3%) operations were done on-pump on the arrested heart, 1908 (40.3%) were performed off-pump, 364 (7.7%) were performed on-pump on the beating heart, and 129 (2.7%) were performed with right heart assist. The preoperative risk factors, operative variables, and postoperative outcomes of the groups were analyzed. Results: Patients selected for on-pump beating heart procedures tended to be sicker with the highest predicted risk of death. We also selected patients who were in cardiogenic shock, in resuscitation, in emergent or salvage status, on dialysis, and with preoperative intra-aortic balloon pump (IABP) use for on-pump beating heart procedures at higher than expected rates. Patients with renal failure with or without dialysis, and those having a history of cerebrovascular accident tended not to be chosen for on-pump arrested heart procedures. Off-pump beating heart procedures were avoided for patients with cardiogenic shock or resuscitation, in emergent or salvage status, and with preoperative IABP use. The mortality rate in these patients was slightly worse in the on-pump beating heart group (4.4%) than in the on-pump arrested heart (3.5%) and off-pump (2.3%) groups (analysis of variance [ANOVA], P =.04). Atrial fibrillation occurred more frequently in both the on-pump beating heart (20.1%) and on-pump arrested heart (23.8%) groups (ANOVA, P <.001). The on-pump groups had higher rates of blood product use and reoperation for bleeding and a prolonged ventilation rate, compared with the other procedures. On-pump patients had a statistically longer length of stay than either off-pump or right heart-assisted patients ( P <.05) and required longer times on the ventilator and in the intensive care unit. Conclusions: Normothermic cardiopulmonary bypass with a beating heart is safe and efficacious and may be the method of choice for patients in cardiogenic shock, requiring resuscitation, or with previous CABG surgery, recent myocardial infarction, a low ejection fraction, or unstable arrhythmias.     

Comparison between leadless and transvenous single-chamber pacemaker therapy in a referral centre for lead extraction

Journal of Interventional Cardiac Electrophysiology - The aim of the study was to compare the long-term clinical and electrical performance of Micra leadless pacemaker with transvenous...     

Ontogeny of hallucal metatarsal rigidity and shape in the rhesus monkey (Macaca mulatta) and chimpanzee (Pan troglodytes)

Life history variables including the timing of locomotor independence, along with changes in preferred locomotor behaviors and substrate use during development, influence how primates use their feet throughout ontogeny. Changes in foot function during development, in particular the nature of how the hallux is used in grasping, can lead to different structural changes in foot bones. To test this hypothesis, metatarsal midshaft rigidity [estimated from the polar second moment of area (J) scaled to bone length] and cross‐sectional shape (calculated from the ratio of maximum and minimum second moments of area, I_max/I_min) were examined in a cross‐sectional ontogenetic sample of rhesus macaques (Macaca mulatta; n = 73) and common chimpanzees (Pan troglodytes; n = 79). Results show the hallucal metatarsal (Mt1) is relatively more rigid (with higher scaled J‐values) in younger chimpanzees and macaques, with significant decreases in relative rigidity in both taxa until the age of achieving locomotor independence. Within each age group, Mt1 rigidity is always significantly higher in chimpanzees than macaques. When compared with the lateral metatarsals (Mt2–5), the Mt1 is relatively more rigid in both taxa and across all ages; however, this difference is significantly greater in chimpanzees. Length and J scale with negative allometry in all metatarsals and in both species (except the Mt2 of chimpanzees, which scales with positive allometry). Only in macaques does Mt1 midshaft shape significantly change across ontogeny, with older individuals having more elliptical cross‐sections. Different patterns of development in metatarsal diaphyseal rigidity and shape likely reflect the different ways in which the foot, and in particular the hallux, functions across ontogeny in apes and monkeys.     

转基因泡桐树叶及其花粉毒性与过敏性试验

目的初步探讨转入抗菌肽基因的泡桐树叶及花粉的毒性与致敏性。方法采用最大灌胃量法进行泡桐树叶的急性经口毒性试验;按照化妆品卫生规范方法进行花粉的皮肤变态反应试验;采用滴鼻和腹腔注射途径给予豚鼠花粉,观察致过敏性哮喘反应。结果转基因泡桐树（树叶）对SPF级Wistar雌性和雄性大鼠急性经口LD50均〉10000mg/kg·BW;转基因泡桐树（花粉）对普通级白毛豚鼠皮肤变态反应积分为0,致敏率为0%,致敏强度为弱,未见皮肤变态反应;转基因泡桐树花粉对豚鼠未见明显过敏性哮喘反应。结论转入抗菌肽基因泡桐树叶属实际无毒级,花粉对豚鼠皮肤致敏性与过敏性哮喘反应均为阴性。     

Molecular cloning and characterization of WD40-repeat protein from Clonorchis sinensis

WD40-repeat proteins have four to eight repeating units flanked by Gly-His (GH) and Trp-Asp (WD) at both termini and folds into a β-propeller. A polypeptide deduced from a Clonorchis sinensis cDNA clone analyzed to have seven WD40-repeats and predicted to form a β-propeller (CsWD1). The CsWD1 protein was expressed stage-specifically in the metacercariae and localized in the tegumental syncytium. The CsWD1 protein is suggested to serve as a platform for interacting partner proteins in the tegumental syncytium of C. sinensis metacercariae.     

Occurrence of Bartonella henselae and Bartonella quintana in a healthy Greek population

The purpose of this study was to determine the prevalence of IgM and IgG antibodies against Bartonella henselae and B. quintana in a healthy Greek population using a commercially available immunofluorescent test (Focus test). Five hundred healthy individuals were divided by sex into four age groups and three groups according to contact with cats. IgM antibodies were not detected in any of the subjects examined, while 99 (19.8%) and 75 (15%) were IgG seropositive to B. henselae and to B. quintana, respectively. No statistical difference in the seropositivity was observed among these groups. The IgG antibody titers ranged from 1/64 to 1/256 for B. henselae and from 1/64 to 1/512 for B. quintana. A high percentage (12.4%) of cross-reactivity between the two species was observed. Our data show that the prevalence of both Bartonella species in Greece is high. However, low IgG antibody levels are not sufficient evidence of active infection.