Is part of the molecular basis of the perineurial barrier function the lack of endogenous carbohydrate-binding proteins?

The sugar part of cellular glycoconjugates and specific endogenous sugar receptors, i.e., lectins, can establish a system of biological recognition based on protein-carbohydrate interactions. An assortment of labelled (neo)glycoproteins, carrying different types of sugar moieties, is synthesized to localize respective sugar receptors. With these tools, the histochemical patterns of endogenous carbohydrate-binding receptors of the epi-, peri-, and endoneurium were analyzed in human sural and accessory nerves and in swine sciatic nerve. This approach is complementary to the application of plant lectins, focusing on endogenous carbohydrate-binding proteins (lectins). In contrast to the epi- and endoneurium, which bound certain types of carbohydrates, such endogenous sugar receptors were histochemically not detectable in the perineurial cells. Moreover, no histochemical reaction was present in the "connective tissue septa" localized in the endoneurium in which the endoneurial vessels were embedded. This common property supplies evidence that these septa are composed of perineurial cells. They may represent a barrier in addition to the capillary endothelium. Our observations suggest histogenetical differences between the cell populations of epi- and endoneurium vs. perineurium. This significant difference in the ability to bind carbohydrate residues, conjugated to a carrier protein, is contradictory to the assumption that perineurial cells and fibroblasts are functional variants of the same cell type. The histochemical patterns of endogenous carbohydrate-binding receptors found in human and swine nerves were similar but not identical, with exception of the perineurium, reflecting phylogenetic differences in the expression of sugar-binding proteins. The absence of specific sugar receptors in perineurial cells, however, seems to be a more general phenomenon.(ABSTRACT TRUNCATED AT 250 WORDS)     

Direct mapping of MHC class II epitopes

Despite technical improvements, the mapping of MHC class II epitopes within complex antigens by genetic or biochemical methods is still laborious and expensive. Here, we describe a simple and fast procedure to directly map T helper cell epitopes within known antigens by bacterial expression cloning. Short antigenic fragments, created by digestion of the coding sequence of the antigen with frequently cutting restriction enzymes, are randomly ligated to the coding sequence of GFP in a bacterial expression vector. Bacteria expressing antigen-GFP fusion proteins are then fed directly to MHC II+ antigen-presenting cells and probed with antigen-specific T cells. Bacterial colonies recognized by T cells are expanded, and the antigenic fragments identified by plasmid extraction and sequence analysis. This direct epitope identification (DEPI) approach offers several advantages. First, bacterial colonies expressing the antigen in frame with GFP are easily detectable by virtue of their green appearance and thereby reduce the screening effort significantly. Second, short antigenic peptides normally unstable in bacteria are highly expressed when fused to GFP. Third, the uniformly high level of expression of short antigenic peptides fused to GFP permits the identification of epitopes even within proteins which are difficult to express in bacteria. Furthermore, by fusing double-stranded oligonucleotides to the GFP gene, crucial amino acids within T cell epitopes may be defined. Thus, this method not only facilitates the identification of T cell epitopes, but also makes it possible to assess the role of individual amino acids for MHC binding or T cell recognition.     

Improving Breastfeeding Environments and Feeding Practices in Family Child Care Homes with the Go NAP SACC Program

Maternal and Child Health Journal - Breastfeeding and responsive feeding are important practices that support the health of infants and women. In the United States, breastfeeding continuation rates...     

Lysosomal phospholipase A2: A novel player in host immunity to Mycobacterium tuberculosis

Phospholipases catalyze the cleavage of membrane phospholipids into smaller bioactive molecules. The lysosomal phospholipase A₂ (LPLA₂) is specifically expressed in macrophages. LPLA₂ gene deletion in mice causes lysosomal phospholipid accumulation in tissue macrophages leading to phospholipidosis. This phenotype becomes most prominent in alveolar macrophages where LPLA₂ contributes to surfactant phospholipid degradation. High expression of LPLA₂ in alveolar macrophages prompted us to investigate its role in host immunity against the respiratory pathogen Mycobacterium tuberculosis, the causative agent of tuberculosis. Here we report that adaptive immune responses to M. tuberculosis were impaired in LPLA₂ deficient mice. Upon aerosol infection with M. tuberculosis, LPLA₂ deficient mice showed enhanced mycobacterial counts but less lung immunopathology and pulmonary inflammatory responses. Compromised T‐cell priming in the lymph nodes was associated with impaired pulmonary T‐cell recruitment and activation. Together with reduced Th1 type cytokine production, these results indicate that LPLA₂ is indispensable for the induction of adaptive T‐cell immunity to M. tuberculosis. Taken together, we identified an unexpected and novel function of a lysosomal phospholipid‐degrading enzyme.     

Biliverdin IX is an endogenous inhibitor of soluble guanylyl cyclase

Heme oxygenase (HO) converts heme to carbon monoxide (CO) and biliverdin IX. CO is a weak activator of soluble guanylyl cyclase (SGC), the enzyme that catalyzes the conversion of GTP to the second messenger cGMP. HO overexpression has recently been shown to inhibit production of cGMP by SGC in vivo. The aim of the present study was to investigate a possible influence of biliverdin IX on SGC activity. Using recombinant alpha(1)/beta(1) isoform of SGC, we show an inhibitory effect of biliverdin IX in the micromolar range both on basal and NO stimulated guanylyl cyclase activity. Bilirubin IX which differs from biliverdin IX in two hydrogen atoms had no effect. Biliverdin IX reduced maximal guanylyl cyclase activity (V(max) values) while it had no effect on the K(M) values indicating unchanged affinity towards the substrate GTP. Concentration response experiments using the NO donor, 2,2-diethyl-1-nitroso-oxyhydrazine (DEA/NO), showed that enzyme activities at maximal DEA/NO concentration were reduced by biliverdin IX. The affinity of the NO-donor, DEA/NO, towards SGC was significantly reduced in the presence of biliverdin IX. Biliverdin IX lowered enzyme activity at maximal activator concentrations of YC-1 and protoporphyrin IX (PPIX) while it had no significant effect on the EC(50) values of these two NO independent activators. The inhibitory effect of biliverdin IX on PPIX activated enzyme activity is not shared by ODQ, which indicates that the inhibitory mechanism of biliverdin IX is different from ODQ.     

Pseudomonads in a new hospital building

Testing the piping of a new hospital showed that the drinking water was contaminated with Pseudomonas aeruginosa. Because of pollution the plumbing system was treated with oxidizing biocides and air-water flushing for several times, unfortunately without stopping the bacterial recontamination. The reasons were firstly stagnation due to dead legs in the piping system, oversized pipes, very low water exchange rates and the installation of a catch water system, secondly low flow velocity because of low water pressure. After these sources of recontamination had been repaired, chlorine dioxide sanitization could be stopped successfully     

Posterior cerebral artery Wada test: sodium amytal distribution and functional deficits

Inadequate sodium amytal delivery to the posterior hippocampus during the intracarotid Wada test has led to development of selective tests. Our purpose was to show the sodium amytal distribution in the posterior cerebral artery (PCA) Wada test and to relate it to functional deficits during the test. We simultaneously injected 80 mg sodium amytal and 14.8 MBq ^{99 m}Tc-hexamethylpropyleneamine oxime (HMPAO) into the P2-segment of the PCA in 14 patients with temporal lobe epilepsy. To show the skull, we injected 116 MBq ^{99 m}Tc-HDP intravenously. Sodium amytal distribution was determined by high-resolution single-photon emission computed tomography (SPECT). In all patients, HMPAO was distributed throughout the parahippocampal gyrus and hippocampus; it was also seen in the occipital lobe in all cases and in the thalamus in 11. Eleven patients were awake and cooperative; one was slightly uncooperative due to speech comprehension difficulties and perseveration. All patients showed contralateral hemianopia during the test. Four patients had nominal dysphasia for 1–3 min. None developed motor deficits or had permanent neurological deficits. Neurological deficits due to inactivation of extrahippocampal areas thus do not grossly interfere with neuropsychological testing during the test. Received: 16 May 2000 Accepted: 21 November 2000     

Dynamic imaging in diagnosis of dysphagia and globus sensation

BACKGROUND: Videofluoroscopy has gained high significance for the evaluation of deglutition disorders. Imaging alone cannot clarify whether the subjective symptom is represented morphologically or whether it is the substrate of a functional disorder. METHOD: The videofluoroscopies of 101 patients with dysphagia (n = 55) and globus pharyngitis (n = 46) were evaluated. Morphologic abnormalities were registered as well as sequential movement patterns. These data were compared with clinical and endoscopic findings. RESULTS: In 87% of the dysphagia and 74% of the globus patients videofluoroscopy revealed pathologic findings. Functional disorders were seen significantly more often than morphologic abnormalities. Highest incidence was found for cricopharyngeal dyskinesia (42%). Hypopharyngeal pouches and degeneration signs of the cervical spine with bolus impression less than 40% are common but functionally not important. Additional esophago-gastroduodenoscopy was pathological in 83% in the dysphagia group and 96% in the globus group. CONCLUSION: Videofluoroscopy is indispensable for the differential diagnosis of dysphagia and globus sensation, especially for the detection of functional disorders in the pharyngoesophageal segment thus documenting the dynamic aspect of deglutition. Videofluoroscopy should be completed by a gastroenterologic examination in order to improve diagnosis.     

Enhanced reduction of myocardial infarct size by combined ACE inhibition and AT(1)-receptor antagonism

The effects of the angiotensin-converting-enzyme inhibitor (ACEI) ramiprilat, the angiotensin II type 1 receptor antagonist (AT(1)A) candesartan, and the combination of both drugs on infarct size (IS) resulting from regional myocardial ischaemia were studied in pigs. Both ACEI and AT(1)A reduce myocardial IS by a bradykinin-mediated process. It is unclear, however, whether the combination of ACEI and AT(1)A produces a more pronounced IS reduction than each of these drugs alone. Forty-six enflurane-anaesthetized pigs underwent 90 min low-flow ischaemia and 120 min reperfusion. Systemic haemodynamics (micromanometer), subendocardial blood flow (ENDO, microspheres) and IS (TTC-staining) were determined. The decreases in left ventricular peak pressure by ACEI (by 9+/-2 (s.e. mean) mmHg), AT(1)A (by 11+/-2 mmHg) or their combination (by 18+/-3 mmHg, P<0.05 vs ACEI and AT(1)A, respectively) were readjusted by aortic constriction prior to ischaemia. With placebo (n=10), IS averaged 20.0+/-3.3% of the area at risk. IS was reduced to 9.8+/-2.6% with ramiprilat (n=10) and 10.6+/-3.1% with candesartan (n=10). Combined ramiprilat and candesartan (n=10) reduced IS to 6.7+/-2.1%. Blockade of the bradykinin-B(2)-receptor with icatibant prior to ACEI and AT(1)A completely abolished the reduction of IS (n=6, 22.8+/-6.1%). The relationship between IS and ischaemic ENDO with placebo was shifted downwards by each ACEI and AT(1)A and further shifted downwards with their combination (P<0.05 vs all groups); icatibant again abolished such downward shift. The combination of ACEI and AT(1)A enhances the reduction of IS following ischaemia/reperfusion compared to a monotherapy by either drug alone; this effect is mediated by bradykinin.