Elevated levels of IgG and IgG4 to Malassezia allergens in atopic eczema patients with IgE reactivity to Malassezia

BACKGROUND: The opportunistic yeast Malassezia is considered to be one of the factors that can contribute to atopic eczema (AE). Elevated serum IgE levels, T-cell proliferation and positive skin prick test (SPT) and atopy patch test (APT) reactions to Malassezia are found among AE patients. METHODS: Sera from 127 AE patients, 14 patients with seborrheic dermatitis (SD) and 33 healthy controls were investigated for IgE and IgG4 to M. sympodialis extract and four recombinant Malassezia allergens; rMala s 1, rMala s 5, rMala s 6, and rMala s 9. In addition, IgG to the recombinant allergens was analyzed. The IgG and IgG4 levels were compared to IgE levels and in vivo reactions (SPT and APT) to Malassezia. RESULTS: AE patients with serum IgE levels >0.35 kU/l to M. sympodialis extract had significantly higher IgG4 levels to M. sympodialis extract than AE patients without detectable serum IgE to M. sympodialis extract, SD patients and healthy controls. Among the AE patients with and without detectable serum IgE to M. sympodialis extract, respectively, there were no differences in IgG4 levels between patients with positive or negative in vivo reactions to M. sympodialis extract. IgG4 to the rMala s allergens was almost exclusively found among patients with IgE to the same allergen. Within the four tested rMala s allergens, most IgG4 reactions were found to rMala s 6, an allergen with homology to cyclophilin. CONCLUSIONS: Elevated serum IgG4 to M. sympodialis extract accompanies elevated serum IgE to the extract. This is further confirmed by the association between IgG/IgG4 and IgE to recombinant Malassezia allergens.     

Formation of 6-nitro-norepinephrine from nitric oxide and norepinephrine in the spinal cord and its role in spinal analgesia

Spinally released norepinephrine is thought to produce analgesia in part by stimulating alpha(2)-adrenergic receptors, which in turn leads to nitric oxide synthesis. Also, nitric oxide is known to react with norepinephrine in vivo in the brain to form 6-nitro-norepinephrine, which inhibits neuronal norepinephrine reuptake. In the present study, we tested the hypothesis that formation of 6-nitro-norepinephrine occurs in the spinal cord and that intrathecal administration of 6-nitro-norepinephrine produces analgesia by stimulating norepinephrine release. 6-Nitro-norepinephrine was present in rat spinal cord tissue and microdialysates of the dorsal horn and intrathecal space. Intrathecal norepinephrine injection increased 6-nitro-norepinephrine. 6-Nitro-norepinephrine also stimulated norepinephrine release in dorsal spinal cord in vitro. Intrathecal injection of 6-nitro-norepinephrine produced antinociception and interacted additively with norepinephrine for antinociception. Spinal noradrenergic nerve destruction increased antinociception from intrathecally injected norepinephrine, but decreased antinociception from 6-nitro-norepinephrine.These results suggest a functional interaction between spinal nitric oxide and norepinephrine in analgesia, mediated in part by formation of 6-nitro-norepinephrine. Stimulation of auto-inhibitory alpha(2)-adrenergic receptors at noradrenergic synapses decreases norepinephrine release. Paradoxically, alpha(2)-adrenergic agonist injection increases and alpha(2)-adrenergic antagonist injection decreases norepinephrine release in the spinal cord. 6-Nitro-norepinephrine may be an important regulator of spinal norepinephrine release and could explain the positive feedback on norepinephrine release after activation of spinal alpha(2)-adrenergic receptors.     

Kinetoplast DNA signatures of Trypanosoma cruzi strains obtained directly from infected tissues.

We report here a polymerase chain reaction (PCR)-based DNA profiling technique that permits Trypanosoma cruzi strain characterization by direct study of infected tissues. This is based on application of a recently developed method of DNA fragment identification, called low-stringency single specific primer PCR (LSSP-PCR), to the study of the variable region of kinetoplast DNA (kDNA) minicircles from T. cruzi Thus, we can translate the intraspecific polymorphism in the nucleotide sequence of kDNA minicircles into a specific and highly reproducible kDNA signature. Comparison with the phenogram obtained by DNA fingerprinting analysis of a set of T. cruzi strains showed good qualitative correlation between the degree of divergence of the LSSP-PCR profiles and the genetic distance between the strains. kDNA signatures of heart tissue from acutely or chronically infected animals revealed perfect concordance with the patterns obtained from cultured parasites for the CL and Colombiana strains but not for the Y strain, which is known to be multiclonal. However, the match was perfect for studies with two clones of the Y strain. We take this as evidence that in some multiclonal strains there is heterogeneity among the clones in the degree of tropism for the heart tissue. Finally, we showed that it is possible to obtain a T. cruzi kDNA signature from the heart of a human patient with chronic Chagasic myocardiopathy. kDNA signatures obtained by LSSP-PCR of sequences amplified from infected tissues constitute a new tool to study the molecular epidemiology of Chagas' disease.     

Mucosal FOXP3-expressing CD4+ CD25high regulatory T cells in Helicobacter pylori-infected patients

Helicobacter pylori chronically colonizes the stomach and duodenum and causes peptic ulcers or gastric adenocarcinoma in 10 to 20% of infected individuals. We hypothesize that the inability of patients to clear H. pylori infections is a consequence of active suppression of the immune response. Here we show that H. pylori-infected individuals have increased frequencies of CD4(+) CD25(high) T cells in both the stomach and duodenal mucosa compared to uninfected controls. These cells have the phenotype of regulatory T cells, as they express FOXP3, a key gene for the development and function of regulatory T cells, as well as high levels of the cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) protein. In contrast, mucosal CD4(+) CD25(low) and CD4(+) CD25(-) cells express little FOXP3 mRNA and low levels of the CTLA-4 protein. Mucosal CD4(+) CD25(high) T cells are present in individuals with asymptomatic H. pylori infections as well as in duodenal ulcer patients. The frequencies of CD4(+) CD25(high) cells are also increased in the stomachs of H. pylori-infected patients with gastric adenocarcinoma, particularly in cancer-affected tissues. These findings suggest that regulatory T cells may suppress mucosal immune responses and thereby contribute to the persistence of H. pylori infections.     

Changes in the surface electromyogram during increasing isometric shoulder forward flexions

Summary When using electromyographic techniques in the evaluation of muscular load it is necessary to determine the mathematical relationship between the torque and the amplitude of the electromyographic signal. Isometric gradually increasing contractions up to 100% MVC can then be used. Often more than linear increases for the amplitude (RMS) — force regression have been reported. The present study was designed to test whether changes in power spectral density function take place during a gradually increasing isometric contraction (duration 10 s). Twenty-two clinically healthy females performed an increasing isometric shoulder forward flexion for 10 s using an isokinetic dynamometer. Electromyographic activity was measured in trapezius, deltoid, infraspinatus and biceps brachii using surface electrodes. Mean torque values were determined together with mean power frequency (MPF) and root mean square values (RMS) from the EMG signals for each 256 ms period. The RMS-torque regressions showed higher regression coefficients during the 6th to 9th sec than during the first 5 s. No significant correlation existed between MPF for the four muscles and the torque. A gradual decrease in MPF was generally found from the 6th s. It is concluded that this decrease in power spectral density function might have contributed to the significantly higher regression coefficient for the RMS torque regression at the high output part of the gradually increasing isometric contraction.     

Biochemical investigation with regard to infection and immunity of Eimeria acervulina in the fowl

Summary Biochemical investigation of the fluid surrounding the sporocysts in the oocysts (oocyst fluid) showed the presence of the common amino acids, -isoaminobutyric acid, glycerol, an unidentified carbohydrate and proteins.Incubation experiments with D-glucose-¹⁴C (U) revealed the presence of enzymes able to convert glucose into lactic acid and other acids. Inside the sporocysts the common amino acids, glycerol and the unidentified carbohydrate were also present, but -isoaminobutyric acid did not occur. The carbohydrate binding protein carboglutelin, carbohydrate phosphate and small amounts of glucose and fructose were mainly found inside the sporocysts.Incubation experiments of oocyst fluid mixed with sporocysts with D-glucose-C¹⁴ (U) demonstrated an interchange between the fluid and the sporocysts. In the oocyst fluid lactic acid was formed, but inside the sporocysts the conversion appeared to stop mainly with formation of carbohydrate phosphate.Incubation experiments of intestinal pieces both from immunized and not-immunized birds with oocyst fluid, sporocysts and labelled glucose showed that a stronger reaction took place in immunized birds than in not-immunized ones.Similar experiments were performed with not-immunized birds at different days after a primary infection. The reaction of the intestinal wall, which seemed quite normal again since day 19 after infection, was comparable with that observed in immunized birds.Finally the hypothesis is given, that in the first part of the intestine of immunized birds compounds are present originating from the first infection. These compounds might enhance the reaction between the oocyst fluid and the present feed to such an extent that the pH decreases and much epithelial cells are pushed off possibly together with the sporocysts. Then leakage of serum proteins might occur.     

Clinical Concept-Based Radiology Reports Classification Pipeline for Lung Carcinoma

Rising incidence and mortality of cancer have led to an incremental amount of research in the field. To learn from preexisting data, it has become important to capture maximum information related to disease type, stage, treatment, and outcomes. Medical imaging reports are rich in this kind of information but are only present as free text. The extraction of information from such unstructured text reports is labor-intensive. The use of Natural Language Processing (NLP) tools to extract information from radiology reports can make it less time-consuming as well as more effective. In this study, we have developed and compared different models for the classification of lung carcinoma reports using clinical concepts. This study was approved by the institutional ethics committee as a retrospective study with a waiver of informed consent. A clinical concept-based classification pipeline for lung carcinoma radiology reports was developed using rule-based as well as machine learning models and compared. The machine learning models used were XGBoost and two more deep learning model architectures with bidirectional long short-term neural networks. A corpus consisting of 1700 radiology reports including computed tomography (CT) and positron emission tomography/computed tomography (PET/CT) reports were used for development and testing. Five hundred one radiology reports from MIMIC-III Clinical Database version 1.4 was used for external validation. The pipeline achieved an overall F1 score of 0.94 on the internal set and 0.74 on external validation with the rule-based algorithm using expert input giving the best performance. Among the machine learning models, the Bi-LSTM_dropout model performed better than the ML model using XGBoost and the Bi-LSTM_simple model on internal set, whereas on external validation, the Bi-LSTM_simple model performed relatively better than other 2. This pipeline can be used for clinical concept-based classification of radiology reports related to lung carcinoma from a huge corpus and also for automated annotation of these reports.

     

Trypanosomicidal kaurane diterpenes from Wedelia paludosa

The diterpenes ent-kaur-16-en-19-oic acid, ent-kaur-9(11),16(17)-dien-19-oic acid and 3 alpha-angeloiloxy-ent-kaur-16-en-19-oic acid were identified as trypanosomicidal compounds of the ethanolic extract from the aerial parts of Wedelia paludosa D.C. (Asteraceae), showing activity up to the lowest dose of 0.68 mg/mL in the in vitro assay against trypomastigotes of T. cruzi, the causative agent of Chagas' disease (American trypanosomiasis). The other isolates, friedelan-3 beta-ol, ent-kaur-16 alpha-ol-19-oic acid, beta-amyrin acetate and (22-E)-stigmasta-5,22-dien-3 beta-ol, were inactive. This is the first report on the trypanosomicidal activity of ent-kaur-9(11),16(17)-dien-19-oic acid and 3 alpha-angeloiloxy-ent-kaur-16-en-19-oic acids; this effect was already known for ent-kaur-16-en-19-oic acid.