Gene conversion is a likely cause of mutation in PKD1

Approximately 70% of the gene responsible for the most common form of autosomal dominant polycystic kidney disease ( PKD1 ) is replicated in several highly homologous copies located more proximally on chromosome 16. We recently have described a novel technique for mutation detection in the duplicated region of PKD1 that circumvents the difficulties posed by these homologs. We have used this method to identify two patients with a nearly identical cluster of base pair substitutions in exon 23. Since pseudogenes are known to be reservoirs for mutation via gene conversion events for a number of other diseases, we decided to test whether these sequence differences in PKD1 could have arisen as a result of this mechanism. Using changes in restriction digest patterns, we were able to show that these sequence substitutions are also present in N23HA, a rodent-human somatic cell hybrid that contains only the PKD1 homologs. Moreover, these changes were also detected in total DNA from several affected and unaffected individuals that did not harbor this mutation in their PKD1 gene copy. This is the first example of gene conversion in PKD1 , and our findings highlight the importance of using gene-specific reagents in defining PKD1 mutations.      

Endoplasmic reticulum chaperone gp96 in macrophages is essential for protective immunity during Gram‐negative pneumonia

Klebsiella pneumoniae is among the most common Gram‐negative bacteria that cause pneumonia. Gp96 is an endoplasmic reticulum chaperone that is essential for the trafficking and function of Toll‐like receptors (TLRs) and integrins. To determine the role of gp96 in myeloid cells in host defence during Klebsiella pneumonia, mice homozygous for the conditional Hsp90b1 allele encoding gp96 were crossed with mice expressing Cre‐recombinase under control of the LysM promoter to generate LysMcre‐Hsp90b1‐flox mice. LysMcre‐Hsp90b1‐flox mice showed absence of gp96 protein in macrophages and partial depletion in monocytes and granulocytes. This was accompanied by almost complete absence of TLR2 and TLR4 on macrophages. Likewise, integrin subunits CD11b and CD18 were not detectable on macrophages, while being only slightly reduced on monocytes and granulocytes. Gp96‐deficient macrophages did not release pro‐inflammatory cytokines in response to Klebsiella and displayed reduced phagocytic capacity independent of CD18. LysMcre‐Hsp90b1‐flox mice were highly vulnerable to lower airway infection induced by K. pneumoniae, as reflected by enhanced bacterial growth and a higher mortality rate. The early inflammatory response in Hsp90b1‐flox mice was characterized by strongly impaired recruitment of granulocytes into the lungs, accompanied by attenuated production of pro‐inflammatory cytokines, while the inflammatory response during late‐stage pneumonia was not dependent on the presence of gp96. Blocking CD18 did not reproduce the impaired host defence of LysMcre‐Hsp90b1‐flox mice during Klebsiella pneumonia. These data indicate that macrophage gp96 is essential for protective immunity during Gram‐negative pneumonia by regulating TLR expression. Copyright © 2015 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Somatic cell hybrid analyses of hematopoietic differentiation

A differentiated cell expresses an entire set of specialized features. Somatic cell hybridization provides a method to examine control of gene regulation. We studied the expression of tissue-specific features in hybrids between human promyelocytes (HL-60) and human Burkitt's lymphoma cells (P3HR-1). Two hybrid lines, HP-1 and HP-2, and 18 hybrid clones were established and confirmed by karyotype, isozyme, and surface antigen analyses. The hybrids extinguished the 10 myeloid (HL- 60) features that we examined including myeloid morphology, histochemistry, and functions that included response to colony- stimulating factor and ability to differentiate to granulocytes or macrophages. In contrast, the hybrids synthesized immunoglobulin and expressed Epstein-Barr nuclear, early, and viral capsid antigens similar to the P3HR-1 lymphoid parental line. Results are contrasted to the findings when P3HR-1 lymphocytes are fused to human erythroid- myeloid cells (K562). Taken together, our results suggest that phenotypic differences between human myeloid and lymphoid cells in the hematopoietic lineage involve mutually exclusive programs and may possibly be mediated by the activity of diffusible, transacting molecules.     

Renal thorium deposition associated with transitional cell carcinoma: radiologic demonstration in two patients

Abdominal radiography, excretory urography, retrograde pyelography, and computed tomography were performed in two patients who had undergone retrograde pyelography with thorium dioxide (Thorotrast) approximately 40 years ago. Both patients developed a transitional cell carcinoma due to suburothelial thorium deposition. Typical thorium densities were demonstrated at CT in the peripelvicalyceal area as well as in retroperitoneal lymph nodes. Elderly patients in whom radiographic examination reveals retained Thorotrast in the kidney should be followed up because of the high risk of renal carcinoma.     

替硝唑软膏的研制与临床应用

目的:研究替硝唑软膏,并应用于临床。方法:参照同类制剂,确定最优组方和制备方法,采用紫外分光光度法测定含量,用比较法进行临床疗效观察。结果:制剂制备简单,含量测定方法简便,结果可靠,经临床应用牙周炎、牙龈炎患者62例,总有效率95.3％。结论:本品是治疗牙周炎、牙龈炎的理想制剂,值得推广应用。     

Reliable and valid NEWS for Chinese seniors: measuring perceived neighborhood attributes related to walking

Background

The effects of the built environment on walking in seniors have not been studied in an Asian context. To examine these effects, valid and reliable measures are needed. The aim of this study was to develop and validate a questionnaire of perceived neighborhood characteristics related to walking appropriate for Chinese seniors (Neighborhood Environment Walkability Scale for Chinese Seniors, NEWS-CS). It was based on the Neighborhood Environment Walkability Scale - Abbreviated (NEWS-A), a validated measure of perceived built environment developed in the USA for adults. A secondary study aim was to establish the generalizability of the NEWS-A to an Asian high-density urban context and a different age group.

Methods

A multidisciplinary panel of experts adapted the original NEWS-A to reflect the built environment of Hong Kong and needs of seniors. The translated instrument was pre-tested on a sample of 50 Chinese-speaking senior residents (65+ years). The final version of the NEWS-CS was interviewer-administered to 484 seniors residing in four selected Hong Kong districts varying in walkability and socio-economic status. Ninety-two participants completed the questionnaire on two separate occasions, 2-3 weeks apart. Test-rest reliability indices were estimated for each item and subscale of the NEWS-CS. Confirmatory factor analysis was used to develop the measurement model of the NEWS-CS and cross-validate that of the NEWS-A.

Results

The final version of the NEWS-CS consisted of 14 subscales and four single items (76 items). Test-retest reliability was moderate to good (ICC > 50 or % agreement > 60) except for four items measuring distance to destinations. The originally-proposed measurement models of the NEWS-A and NEWS-CS required 2-3 theoretically-justifiable modifications to fit the data well.

Conclusions

The NEWS-CS possesses sufficient levels of reliability and factorial validity to be used for measuring perceived neighborhood environment in Chinese seniors. Further work is needed to assess its construct validity and generalizability to other Asian locations. In general, the measurement model of the original NEWS-A was generalizable to this study context, supporting the feasibility of cross-country and age-group comparisons of the effect of the neighborhood environment on walking using the NEWS-A as a tool to measure the perceived built environment.