Folate receptors targeted to clathrin-coated pits cannot regulate vitamin uptake.

Potocytosis is an endocytic process that is specialized for the internalization of small molecules. Recent studies on the uptake of 5-methyltetrahydrofolate by the folate receptor have suggested that the glycosyl-phosphatidylinositol anchor on this protein causes it to cluster and be internalized by caveolae instead of coated pits. To test this hypothesis directly, we have constructed a chimeric folate receptor that has the glycosyl-phosphatidylinositol anchor replaced with the transmembrane domain and cytoplasmic tail of the low density lipoprotein receptor. The cells with wild-type receptors delivered 5-methyltetrahydrofolate to the cytoplasm more rapidly than did cells expressing the chimeric receptor. This suggests that efficient delivery to the cytoplasm depends on caveolae. In sharp contrast to cells with wild-type folate receptors, cells internalizing folate by clathrin-coated pits were unable to decrease vitamin uptake when they were either folate replete or confluent.     

Nasal-continuous positive airway pressure reduces pulmonary morbidity and length of hospital stay following thoracoabdominal aortic surgery

STUDY OBJECTIVES: Patients who undergo surgical repair of thoracoabdominal aortic aneurysms have a high risk for the development of respiratory complications, which cause significant postoperative morbidity and prolong hospitalization, compared to patients who undergo other types of surgery. We studied whether prophylactic noninvasive application of nasal continuous positive airway pressure (nCPAP) administered via a facemask immediately after extubation may reduce pulmonary morbidity and shorten the length of hospitalization. DESIGN: Prospective randomized clinical trial. SETTING: Surgical ICU of a university hospital. PATIENTS: Fifty-six patients following elective prosthetic replacement of the thoracoabdominal aorta, of whom 6 patients were excluded because they had received prolonged mechanical ventilation. INTERVENTIONS: Following extubation in the ICU, nCPAP was applied for 12 to 24 h at an airway pressure of 10 cm H2O to patients in the study group (n = 25). Subjects in the control group (n = 25) received standard treatment including intermittent nCPAP (10 cm H2O for 10 min) every 4 h. MEASUREMENTS AND RESULTS: In the study group, nCPAP was applied for a mean (+/- SD) duration of 23 +/- 3 h at an airway pressure of 10 +/- 1 cm H2O, which improved pulmonary oxygen transfer without altering hemodynamics (ie, heart rate, mean arterial BP, and central venous pressure). The application of nCPAP was associated with fewer pulmonary complications (Pa(O2)/fraction of inspired oxygen [F(IO2)] <100, atelectasis, pneumonia, reintubation rate) compared to the control group (7 of 25 patients vs 24 of 25 subjects, respectively; p = 0.019). The mean duration of intensive care treatment tended to be shorter in the study group compared to the control group (8 +/- 1 vs 12 +/- 2 days, respectively; difference not significant), while the mean length of hospital stay was shorter with nCPAP therapy (22 +/- 2 vs 34 +/- 5 days, respectively; p = 0.048). CONCLUSIONS: The prophylactic application of nCPAP at airway pressures of 10 cm H2O significantly reduced pulmonary morbidity and length of hospital stay following the surgical repair of thoracoabdominal aortic aneurysms. Thus, it can be recommended as a standard treatment procedure for this patient group.     

Detection of Aspergillus species in blood and bronchoalveolar lavage samples from immunocompromised patients by means of 2-step polymerase chain reaction: clinical results.

Bronchoalveolar lavage (BAL) samples from 67 patients who were at high risk for invasive aspergillosis were examined using a recently developed 2-step polymerase chain reaction (PCR) that detects 相似文献   

Hepatitis E in liver transplant recipients in the Rhône-Alpes region in France

Purpose

In developed countries, hepatitis E virus (HEV) is considered an emerging pathogen, but prevalence seems highly variable according to previous European studies. As HEV can lead to chronic infections in immunosuppressed patients, it is thus essential to evaluate the prevalence and incidence of this infection.

Methods

We determined retrospectively, in a cohort of 206 pediatric and adult liver transplant recipients from the Rhône-Alpes region in France, pre-transplant anti-HEV-IgG prevalence and incidence of HEV infections during post-transplant follow-up (HEV IgG and IgM ± HEV-RNA).

Results

Transplantations were carried out between 2005 and 2012 and mean post-transplant follow-up was 32.8 months. Global pre-transplant prevalence of anti-HEV IgG was 29 %, increasing regularly with age from 7 % for children under 15 to 49 % for patients older than 60. From the 142 seronegative patients before transplant, 11 seroconversions (7.7 %) were observed during follow-up (incidence of 2.83 cases per 100 person-years). HEV RNA—tested at transaminases peak or randomly—was detected in only one case of seroconversion. For at least 2 HEV-seropositive patients, who had negative RNAemia before transplantation, viral RNA was detected chronically during follow-up, suggesting reinfection with HEV.

Conclusion

Acute infections were largely more frequent than chronic infections and were asymptomatic or misdiagnosed, suggesting that liver transplant patients may not be particularly prone to developing severe HEV hepatitis. In addition, the presence of IgG anti-HEV may not protect against re-infection. Serological testing, therefore, appears to be of limited interest for the diagnosis of HEV infections in liver transplant recipients.     

Functional relevance of NLRP3 inflammasome-mediated interleukin (IL)-1β during acute allergic airway inflammation

Overall asthmatic symptoms can be controlled with diverse therapeutic agents. However, certain symptomatic individuals remain at risk for serious morbidity and mortality, which prompts the identification of novel therapeutic targets and treatment strategies. Thus, using an adjuvant-free T helper type 2 (Th2) murine model, we have deciphered the role of interleukin (IL)-1 signalling during allergic airway inflammation (AAI). Because functional IL-1β depends on inflammasome activation we first studied asthmatic manifestations in specific inflammasome-deficient [NACHT, LRR and PYD domains-containing protein 3 (NLRP3^−/−) and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC^−/−)] and IL-1 receptor type 1^−/− (IL-1R1^−/−) mice on the BALB/c background. To verify the onset of disease we assessed cellular infiltration in the bronchial regions, lung pathology, airway hyperresponsiveness and ovalbumin (OVA)-specific immune responses. In the absence of NLRP3 inflammasome-mediated IL-1β release all symptoms of AAI were reduced, except OVA-specific immunoglobulin levels. To address whether manipulating IL-1 signalling reduced asthmatic development, we administered the IL-1R antagonist anakinra (Kineret®) during critical immunological time-points: sensitization or challenge. Amelioration of asthmatic symptoms was only observed when anakinra was administered during OVA challenge. Our findings indicate that blocking IL-1 signalling could be a potential complementary therapy for allergic airway inflammation.     

Effect of channel mutations on the uptake and release of the retinal ligand in opsin

In the retinal binding pocket of rhodopsin, a Schiff base links the retinal ligand covalently to the Lys296 side chain. Light transforms the inverse agonist 11-cis-retinal into the agonist all-trans-retinal, leading to the active Meta II state. Crystal structures of Meta II and the active conformation of the opsin apoprotein revealed two openings of the 7-transmembrane (TM) bundle towards the hydrophobic core of the membrane, one between TM1/TM7 and one between TM5/TM6, respectively. Computational analysis revealed a putative ligand channel connecting the openings and traversing the binding pocket. Identified constrictions within the channel motivated this study of 35 rhodopsin mutants in which single amino acids lining the channel were replaced. 11-cis-retinal uptake and all-trans-retinal release were measured using UV/visible and fluorescence spectroscopy. Most mutations slow or accelerate both uptake and release, often with opposite effects. Mutations closer to the Lys296 active site show larger effects. The nucleophile hydroxylamine accelerates retinal release 80 times but the action profile of the mutants remains very similar. The data show that the mutations do not probe local channel permeability but rather affect global protein dynamics, with the focal point in the ligand pocket. We propose a model for retinal/receptor interaction in which the active receptor conformation sets the open state of the channel for 11-cis-retinal and all-trans-retinal, with positioning of the ligand at the active site as the kinetic bottleneck. Although other G protein-coupled receptors lack the covalent link to the protein, the access of ligands to their binding pocket may follow similar schemes.     

Accuracy of cone beam computed tomography in assessing peri-implant bone defect regeneration: a histologically controlled study in dogs

Objective: To assess the accuracy of cone‐beam computed tomography (CBCT) in terms of buccal bone‐wall configuration and peri‐implant bone defect regeneration after guided bone regeneration (GBR). Material and methods: Titanium implants were inserted into standardized box‐shaped defects in the mandible of 12 foxhounds. Defects of one side were augmented following the principle of GBR, while the other side was left untreated. Radiological evaluation was performed using CBCT and compared with histomorphometrical measurements of the respective site serving as a validation method. Results: Non‐augmented control sites providing a horizontal bone width (BW) of<0.5 mm revealed a significantly lower accuracy between the radiological and the histological evaluation of the buccal defect depth (1.93 ± 1.59 mm) compared with the group providing a BW of >0.5 mm (0.7 ± 0.7 mm) (P<0.05, Mann–Whitney U‐test). In GBR‐treated defects, the subgroup <0.5 mm (1.49 ± 1.29 mm) revealed a significantly higher difference between CBCT and histology compared with >0.5 mm (0.82 ± 1.07) (P>0.05, Mann–Whitney U‐test). However, a radiological discrimination between original bone, integrated and non‐integrated bone substitute material was not reliable. Additionally, it was found that a minimum buccal BW of 0.5 mm was necessary for the detection of bone in radiology. Conclusion: The evaluation of peri‐implant bone defect regeneration by means of CBCT is not accurate for sites providing a BW of <0.5 mm. Moreover, a safe assessment of the success of the GBR technique is not possible after the application of a radiopaque bone substitute material. To cite this article:
Fienitz T, Schwarz F, Ritter L, Dreiseidler T, Becker J, Rothamel D. Accuracy of cone beam computed tomography in assessing peri‐implant bone defect regeneration: a histologically controlled study in dogs.
Clin. Oral Impl. Res. 23 , 2012; 882–887.
doi: 10.1111/j.1600‐0501.2011.02232.x