细菌Ⅲ型分泌系统

1 .细菌分泌机制概述致病细菌和宿主细胞相互作用的特点是将致病因子定位到细菌表面或分泌到胞外。虽然其分泌的细菌蛋白质数量和种类都很多且具有范围很广的作用 (包括蛋白质水解、溶血、细胞毒性、蛋白质的磷酸化和去磷酸化 ) ,但是只存在几种途径把这些蛋白质从细菌的细胞质运输到胞外。在革兰氏阴性细菌中已发现四种蛋白质分泌途径 ,称为Ｉ、ＩＩ、ＩＩＩ、ＩＶ型分泌机制。另外 ,第五种大分子分泌途径和质粒的接合转移有关。根癌农杆菌Ｔ ＤＮＡ的转移 ,百日咳杆菌毒素的分泌 ,称之为Ｖ型分泌 ,这方面的研究很少[1] 。ＩＩ型和ＩＶ型是…     

Immunohistochemical Video-microdensitometry of Myocardial Collagen Type I and Type III

Collagen is an essential part of the cardiac interstitium. Collagen subtypes, their location, total amount and the architecture of the fibrillar network are of functional importance. Architecture in terms of density of the fibrillar network is assumed to be reflected by the intensity of immunohistochemical staining of collagen. The aim of this study was to evaluate a video-based microdensitometric method for quantifying density expressed as absorbance of collagen subtypes I and III stained with an indirect immunoperoxidase method in myectomy specimens of patients with hypertrophic obstructive cardiomyopathy. Various factors influencing the immunohistochemical staining product and the technical properties of the image analysis system were investigated. Linearity between collagen concentration and the absorbance of the immunohistochemical staining product was demonstrated for collagen I using a dot-blot technique. Immunohistochemical collagen staining and density measure ment were easily reproducible. The cardiac disability of the patients was assessed according to the New York Heart Association (NYHA) criteria. There was a significant increase in collagen type I density with higher NYHA class, whereas no significant association was found for total collagen area fraction. Thus, video-based microdensitometry gives further insight into the structural remodelling of myocardial collagens and reveals their significance in the process of heart failure in hypertrophic cardiomyopathy.     

Blood Metals Concentration in Type 1 and Type 2 Diabetics

Mechanisms for the onset of diabetes and the development of diabetic complications remain under extensive investigations. One of these mechanisms is abnormal homeostasis of metals, as either deficiency or excess of metals, can contribute to certain diabetic outcomes. Therefore, this paper will report the blood levels of chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), mercury (Hg), nickel (Ni), lead (Pb), selenium (Se), and zinc (Zn) in subjects with type 1 diabetes (n?=?192, mean age 48.8 years, mean disease duration 20.6 years), type 2 diabetes (n?=?68, mean age 68.4 years, mean disease duration 10.2 years), and in control subjects (n?=?59, mean age 57.2 years), and discuss the results indicating their possible role in diabetes. The metal concentrations were measured by sector field inductively coupled plasma mass spectrometry after microwave-induced acid digestion of blood samples. The accuracy was checked using a blood-based certified reference material, and recoveries of all elements were in the range of 92–101 % of certified values. Type 1 diabetes was found to be associated with Cr (p?=?0.02), Mn (p?<?0.001), Ni (p?<?0.001), Pb (p?=?0.02), and Zn (p?<?0.001) deficiency, and type 2 diabetes with Cr (p?=?0.014), Mn (p?<?0.001), and Ni (p?<?0.001) deficiency. These deficiencies were appreciated also subdividing the understudied patients for gender and age groups. Furthermore, in type 1 diabetes, there was a positive correlation between Pb and age (p?<?0.001, ρ?=?0.400) and Pb and BMI (p?<?0.001, ρ?=?0.309), while a negative correlation between Fe and age (p?=?0.002, ρ?=??0.218). In type 2 diabetes, there was a negative correlation between Fe and age (p?=?0.017, ρ?=??0.294) and Fe and BMI (p?=?0.026, ρ?=??0.301). Thus, these elements may play a role in both forms of diabetes and combined mineral supplementations could have beneficial effects.     

Type III Hyperlipoproteinaemia

Eighteen patients with type III hyperlipoproteinaemia, diagnosed on the basis of skin lesions, serum lipids, and lipoprotein electrophoresis, have been fully investigated over a period of 15 years. The incidence of coronary artery disease was only slightly increased, and was not increased at all among first-degree relatives. Peripheral occlusive arterial disease was probably more common. An increased incidence of carbohydrate intolerance was found in neither the patients nor their relatives. The effects of treatment on the skin were uniformly good.     

IL-10 Dependent Suppression of Type 1, Type 2 and Type 17 Cytokines in Active Pulmonary Tuberculosis

Background

Although Type 1 cytokine responses are considered protective in pulmonary tuberculosis (PTB), their role as well as those of Type 2, 17 and immunoregulatory cytokines in tuberculous lymphadenitis (TBL) and latent tuberculosis (LTB) have not been well studied.

Aim and Methods

To identify cytokine responses associated with pulmonary tuberculosis (TB), TB lymphadenitits and latent TB, we examined mycobacterial antigen-specific immune responses of PTB, TBL and LTB individuals. More specifically, we examined ESAT-6 and CFP-10 induced Type 1, Type 2 and Type 17 cytokine production and their regulation using multiplex ELISA.

Results

PTB individuals exhibited a significantly lower baseline as well as antigen-specific production of Type 1 (IFNγ, TNFα and IL-2); Type 2 (IL-4) and Type 17 (IL-17A and IL-17F) cytokines in comparison to both TBL and LTB individuals. TBL individuals exhibited significantly lower antigen-specific IFNγ responses alone in comparison to LTB individuals. Although, IL-10 levels were not significantly higher, neutralization of IL-10 during antigen stimulation resulted in significantly enhanced production of IFNγ, IL-4 and IL-17A in PTB individuals, indicating that IL-10 mediates (at least partially) the suppression of cytokine responses in PTB.

Conclusion

Pulmonary TB is characterized by an IL-10 dependent antigen-specific suppression of Type 1, Type 2 and Type 17 cytokines, reflecting an important association of these cytokines in the pathogenesis of active TB.     

Genetic Relatedness of Type 1 and Type 2 Herpes Simplex Viruses

The extent of homology between herpes simplex virus(1) and(2) (HSV-1 and HSV-2) deoxyribonucleic acid (DNA) was measured in two ways: (i) by determination of the relative rate of hybridization of labeled HSV-1 and HSV-2 DNA to excess unlabeled HSV-1 or HSV-2 DNA immobilized on filters and (ii) by determination of the rate of hybridization of labeled HSV-1 and HSV-2 DNA to excess unlabeled HSV-1 or HSV-2 DNA in solution. Approximately 40% of HSV-1 and HSV-2 DNA is homologous at hybridization temperatures 25 C below the melting temperature (T(m)) of HSV DNA (liquid-filter annealing). Lowering the temperature to 34 C below the T(m) increased the extent of homology to 46% (liquid annealing). The extent of base-pairing in HSV-1-HSV-2 heteroduplex DNA was determined by thermal chromatography on hydroxyapatite. Heteroduplexes of HSV-1 and HSV-2 DNA eluted in a single peak whose midpoint (Te(50)) was 10 C below that of the homoduplex. Conspicuously absent were heteroduplexes that eluted at more than 15 C below the Te(50) of the homoduplex. The data indicate the existence of a variable region of DNA (54%) with very little, if any, homology and an invariable region (46%) with relatively good (85%) matching of base pairs.     

Comparison of Coding Capabilities of Type I and Type II Neurons

We consider the dependence of information transfer by neurons on the Type I vs. Type II classification of their dynamics. Our computational study is based on Type I and II implementations of the Morris-Lecar model. It mainly concerns neurons, such as those in the auditory or electrosensory system, which encode band-limited amplitude modulations of a periodic carrier signal, and which fire at random cycles yet preferred phases of this carrier. We first show that the Morris-Lecar model with additive broadband noise ("synaptic noise") can exhibit such firing patterns with either Type I or II dynamics, with or without amplitude modulations of the carrier. We then compare the encoding of band-limited random amplitude modulations for both dynamical types. The comparison relies on a parameter calibration that closely matches firing rates for both models across a range of parameters. In the absence of synaptic noise, Type I performs slightly better than Type II, and its performance is optimal for perithreshold signals. However, Type II performs well over a slightly larger range of inputs, and this range lies mostly in the subthreshold region. Further, Type II performs marginally better than Type I when synaptic noise, which yields more realistic baseline firing patterns, is present in both models. These results are discussed in terms of the tuning and phase locking properties of the models with deterministic and stochastic inputs.     

Harnessing Type I and Type III CRISPR-Cas systems for genome editing

CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) systems are widespread in archaea and bacteria, and research on their molecular mechanisms has led to the development of genome-editing techniques based on a few Type II systems. However, there has not been any report on harnessing a Type I or Type III system for genome editing. Here, a method was developed to repurpose both CRISPR-Cas systems for genetic manipulation in Sulfolobus islandicus, a thermophilic archaeon. A novel type of genome-editing plasmid (pGE) was constructed, carrying an artificial mini-CRISPR array and a donor DNA containing a non-target sequence. Transformation of a pGE plasmid would yield two alternative fates to transformed cells: wild-type cells are to be targeted for chromosomal DNA degradation, leading to cell death, whereas those carrying the mutant gene would survive the cell killing and selectively retained as transformants. Using this strategy, different types of mutation were generated, including deletion, insertion and point mutations. We envision this method is readily applicable to different bacteria and archaea that carry an active CRISPR-Cas system of DNA interference provided the protospacer adjacent motif (PAM) of an uncharacterized PAM-dependent CRISPR-Cas system can be predicted by bioinformatic analysis.     

nifH Sequences and Nitrogen Fixation in Type I and Type II Methanotrophs

Some methane-oxidizing bacteria (methanotrophs) are known to be capable of expressing nitrogenase and utilizing N₂ as a nitrogen source. However, no sequences are available for nif genes in these strains, and the known nitrogen-fixing methanotrophs are confined mainly to a few genera. The purpose of this work was to assess the nitrogen-fixing capabilities of a variety of methanotroph strains. nifH gene fragments from four type I methanotrophs and seven type II methanotrophs were PCR amplified and sequenced. Nitrogenase activity was confirmed in selected type I and type II strains by acetylene reduction. Activities ranged from 0.4 to 3.3 nmol/min/mg of protein. Sequence analysis shows that the nifH sequences from the type I and type II strains cluster with nifH sequences from other gamma proteobacteria and alpha proteobacteria, respectively. The translated nifH sequences from three Methylomonas strains show high identity (95 to 99%) to several published translated environmental nifH sequences PCR amplified from rice roots and a freshwater lake. The translated nifH sequences from the type II strains show high identity (94 to 99%) to published translated nifH sequences from a variety of environments, including rice roots, a freshwater lake, an oligotrophic ocean, and forest soil. These results provide evidence for nitrogen fixation in a broad range of methanotrophs and suggest that nitrogen-fixing methanotrophs may be widespread and important in the nitrogen cycling of many environments.     

Type II restriction--modification systems

The genes for increasing numbers of type II restriction and modification enzymes are being cloned and characterized. The gene arrangements and enzyme sequences are quite diverse in spite of the biochemical similarities of the enzymes.