Cytosolic malate dehydrogenase regulates senescence in human fibroblasts

Carbohydrate metabolism changes during cellular senescence. Cytosolic malate dehydrogenase (MDH1) catalyzes the reversible reduction of oxaloacetate to malate at the expense of reduced nicotinamide adenine dinucleotide (NADH). Here, we show that MDH1 plays a critical role in the cellular senescence of human fibroblasts. We observed that the activity of MDH1 was reduced in old human dermal fibroblasts (HDFs) [population doublings (PD) 56], suggesting a link between decreased MDH1 protein levels and aging. Knockdown of MDH1 in young HDFs (PD 20) and the IMR90 human fibroblast cell line resulted in the appearance of significant cellular senescence features, including senescence-associated β-galactosidase staining, flattened and enlarged morphology, increased population doubling time, and elevated p16^INK4A and p21^CIP1 protein levels. Cytosolic NAD/NADH ratios were decreased in old HDFs to the same extent as in MDH1 knockdown HDFs, suggesting that cytosolic NAD depletion is related to cellular senescence. We found that AMP-activated protein kinase, a sensor of cellular energy, was activated in MDH1 knockdown cells. We also found that sirtuin 1 (SIRT1) deacetylase, a controller of cellular senescence, was decreased in MDH1 knockdown cells. These results indicate that the decrease in MDH1 and subsequent reduction in NAD/NADH ratio, which causes SIRT1 inhibition, is a likely carbohydrate metabolism-controlled cellular senescence mechanism.     

A 12-Mb Complete Coverage BAC Contig Map in Human Chromosome 16p13.1–p11.2

We have constructed a complete coverage BAC contig map that spans a 12-Mb genomic segment in the human chromosome 16p13.1-p11.2 region. The map consists of 68 previously mapped STSs and 289 BAC clones, 51 of which-corresponding to a total of 7.721 Mb of genomic DNA-have been sequenced, and provides a high resolution physical map of the region. Contigs were initially built based mainly on the analysis of STS contents and restriction fingerprint patterns of the clones. To close the gaps, probes derived from BAC clone ends were used to screen deeper BAC libraries. Clone end sequence data obtained from chromosome 16-specific BACs, as well as from public databases, were used for the identification of BACs that overlap with fully sequenced BACs by means of sequence match. This approach allowed precise alignment of clone overlaps in addition to restriction fingerprint comparison. A freehand contig drawing software tool was developed and used to manage the map data graphically and generate a real scale physical map. The map we present here is approximately 3.5 x deep and provides a minimal tiling path that covers the region in an array of contigous, overlapping BACs.     

Prevalence of sensitivity to cockroach allergens and IgE cross- reactivity between cockroach and house dust mite allergens in Chinese patients with allergic rhinitis and asthma

Background Cockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of IgE between cockroach and house dust mite allergen in Chinese patients.Methods The cockroach sensitization pattern was based on a skin prick test （SPT） obtained from a national multicenter prevalence study, in which 6304 patients from 25 allergy centers across China participated. Factors, including different regions of China, age, gender and the correlations between the American and German cockroaches and house dust mite Der p were investigated. Eighteen out of 1236 clinical sera from south China were selected to perform the cross-inhibition assay between house dust mites and cockroaches.Results Totally 25.7% of patients were SPT positive to the American cockroach （Periplaneta Americana, Per a） and 18.7% SPT positive to the German cockroach （Blattella germanica, Bla g）. The prevalence of positive cockroach SPT was higher in southern than in northern China, higher in adults than in children, and higher in males than in females.Patients had relatively low levels of cockroach SPT reactions, mainly class 1 or 2. Of the SPT positive cockroach patients,88% were also SPT positive to house dust mite （Dermatophagoides pteronyssinus, Der p）. An IgE cross-inhibition study confirmed that Der P sensitization could cause false positive SPT reactions against cockroach.Conclusions A relatively high prevalence of cockroach sensitivity was found in mainland of China. However, a cross-inhibition study showed that only a small number of patients appear to have Bla g and/or Per a as primary sensitizing source. The importance of cockroaches as a risk factor for sensitization and triggers of allergic symptoms in mainland of China needs to be further investigated.     

cAMP-inducible chloride conductance in mouse fibroblast lines stably expressing the human cystic fibrosis transmembrane conductance regulator.

A cAMP-inducible chloride permeability has been detected in mouse fibroblast (L cell) lines upon stable integration of a full-length cDNA encoding the human cystic fibrosis transmembrane conductance regulator (CFTR). As indicated by a Cl(-)-indicator dye, the Cl- permeability of the plasma membrane increases by 10- to 30-fold within 2 min after treatment of the cells with forskolin, an activator of adenylyl cyclase. The properties of the conductance are similar to those described in secretory epithelial cells; the whole-cell current-voltage relationship is linear and there is no evidence of voltage-dependent inactivation or activation. In contrast, this cAMP-dependent Cl- flux is undetectable in the untransfected cells or cells harboring defective cDNA constructs, including one with a phenylalanine deletion at amino acid position 508 (delta F508), the most common mutation causing cystic fibrosis. These observations are consistent with the hypothesis that the CFTR is a cAMP-dependent Cl- channel. The availability of a heterologous (nonepithelial) cell type expressing the CFTR offers an excellent system to understand the basic mechanisms underlying this CFTR-associated ion permeability and to study the structure and function of the CFTR.     

Histopathological predictors of progression-free survival in atypical meningioma: a single-center retrospective cohort and meta-analysis

Brain Tumor Pathology - To determine the prognostic significance of histopathological features included in the diagnostic criteria of atypical meningioma for progression-free survival (PFS). We...     

Species distribution and in vitro antifungal susceptibility patterns of 75 clinical isolates of Fusarium spp. from northern Italy

Fusarium isolates from 75 Italian patients were identified by molecular methods, and their susceptibilities to antifungals were tested in vitro. Fusarium verticillioides was the species most frequently isolated from deep-seated infections, and F. solani was the species most frequently isolated from superficial infections. F. solani isolates showed high azole MICs, while F. verticillioides isolates showed low posaconazole MICs.