Herbal remedies improve the strength of repairing ligament in a rat model.

Herbal remedies have been reported to be effective in controlling inflammation for acute soft tissue injuries. There exist, however, no reports of their effects on collagen production and remodeling; thus mechanical strength studies of the tissues have not been reported. This study tested the effects of a herbal remedy on the strength of healing medial collateral ligaments (MCL) in rats. Sixteen rats receiving surgical transection to their right MCLs and eight receiving sham operation were tested. Eight of the MCL-injured animals were treated with an adhesive herbal plaster application to their right knees, while the other eight in the MCL injured group and the sham group were treated with plain adhesive plaster to their right knees. The MCLs were harvested and tested at either 3 or 6 weeks post-operation. The ultimate tensile strength (UTS) and stiffness normalized to the uninjured side of each animal of the herb and sham groups were significantly larger than those of the control at both 3 and 6 weeks (p = 0.001). No significant difference was found in stiffness between the herb and sham groups (p > 0.05). We concluded that the herbal remedy improves the UTS and stiffness of repairing MCLs at 3 and 6 weeks after injury.     

Diabetogenic action of human growth hormone. Synthesis and activity of C-terminal fragments.

Three peptides corresponding to the C-terminal region of human growth hormone have been synthesized by the solid-phase method: HGH-(177--191), HGH-(178--191) and HGH-(179--191). The diabetogenic activities of these synthetic peptides are reported. The data indicate that extension of HGH-(179--191) at its NH2-terminus is required for in vivo activity. The reduced and S-carbamidomethylated form of HGH-(177--191) was also active, indicating that the disulphide bond is possibly not a prerequisite for biological activity.     

Spatial Dependency of Tuberculosis Incidence in Taiwan

Tuberculosis (TB) disease can be caused by either recent transmission from infectious patients or reactivation of remote latent infection. Spatial dependency (correlation between nearby geographic areas) in tuberculosis incidence is a signature for chains of recent transmission with geographic diffusion. To understand the contribution of recent transmission in the TB endemic in Taiwan, where reactivation has been assumed to be the predominant mode of pathogenesis, we used spatial regression analysis to examine whether there was spatial dependency between the TB incidence in each township and in its neighbors. A total of 90,661 TB cases from 349 townships in 2003–2008 were included in this analysis. After adjusting for the effects of confounding socioeconomic variables, including the percentages of aboriginals and average household income, the results show that the spatial lag parameter remains positively significant (0.43, p<0.001), which indicates that the TB incidences of neighboring townships had an effect on the TB incidence in each township. Townships with substantial spatial spillover effects were mainly located in the northern, western and eastern parts of Taiwan. Spatial dependency implies that recent transmission plays a significant role in the pathogenesis of TB in Taiwan. Therefore, in addition to the current focus on improving the cure rate under directly observed therapy programs, more resource need to be allocated to active case finding in order to break the chain of transmission.     

Computer‐aided assessment of the chemokine receptors CXCR3, CXCR4 and CXCR7 expression in gallbladder carcinoma

Gallbladder carcinoma (GBC) is a vicious and invasive disease. The major challenge in the clinical treatment of GBC is the lack of a suitable prognosis method. Chemokine receptors such as CXCR3, CXCR4 and CXCR7 play vital roles in the process of tumour progression and metastasis. Their expression levels and distribution are proven to be indicative of the progression of GBC, but are hard to be decoded by conventional pathological methods, and therefore, not commonly used in the prognosis of GBC. In this study, we developed a computer‐aided image analysis method, which we used to quantitatively measure the expression levels of CXCR3, CXCR4 and CXCR7 in the nuclei and cytoplasm of glandular and interstitial cells from a cohort of 55 GBC patients. We found that CXCR3, CXCR4 and CXCR7 expressions are associated with the clinicopathological variables of GBC. Cytoplasmic CXCR3, nuclear CXCR7 and cytoplasmic CXCR7 were significant predictive factors of histology invasion, whereas cytoplasmic CXCR4 and nuclear CXCR4 were significantly correlated with T and N stage and were associated with the overall survival and disease‐free survival. These results suggest that the quantification and localisation of CXCR3, CXCR4 and CXCR7 expressions in different cell types should be considered using computer‐aided assessment to improve the accuracy of prognosis in GBC.     

Elevated intracellular cyclic-di-GMP level in Shewanella oneidensis increases expression of c-type cytochromes

Electrochemically active biofilms are capable of exchanging electrons with solid electron acceptors and have many energy and environmental applications such as bioelectricity generation and environmental remediation. The performance of electrochemically active biofilms is usually dependent on c-type cytochromes, while biofilm development is controlled by a signal cascade mediated by the intracellular secondary messenger bis-(3ʹ-5ʹ) cyclic dimeric guanosine monophosphate (c-di-GMP). However, it is unclear whether there are any links between the c-di-GMP regulatory system and the expression of c-type cytochromes. In this study, we constructed a S. oneidensis MR-1 strain with a higher cytoplasmic c-di-GMP level by constitutively expressing a c-di-GMP synthase and it exhibited expected c-di-GMP-influenced traits, such as lowered motility and increased biofilm formation. Compared to MR-1 wild-type strain, the high c-di-GMP strain had a higher Fe(III) reduction rate (21.58 vs 11.88 pM of Fe(III)/h cell) and greater expression of genes that code for the proteins involved in the Mtr pathway, including CymA, MtrA, MtrB, MtrC and OmcA. Furthermore, single-cell Raman microspectroscopy (SCRM) revealed a great increase of c-type cytochromes in the high c-di-GMP strain as compared to MR-1 wild-type strain. Our results reveal for the first time that the c-di-GMP regulation system indirectly or directly positively regulates the expression of cytochromes involved in the extracellular electron transport (EET) in S. oneidensis, which would help to understand the regulatory mechanism of c-di-GMP on electricity production in bacteria.     

Inhibition of Cdc42 activity extends lifespan and decreases circulating inflammatory cytokines in aged female C57BL/6 mice

Cdc42 is a small RhoGTPase regulating multiple functions in eukaryotic cells. The activity of Cdc42 is significantly elevated in several tissues of aged mice, while the Cdc42 gain‐of‐activity mouse model presents with a premature aging‐like phenotype and with decreased lifespan. These data suggest a causal connection between elevated activity of Cdc42, aging, and reduced lifespan. Here, we demonstrate that systemic treatment of aged (75‐week‐old) female C57BL/6 mice with a Cdc42 activity‐specific inhibitor (CASIN) for 4 consecutive days significantly extends average and maximum lifespan. Moreover, aged CASIN‐treated animals displayed a youthful level of the aging‐associated cytokines IL‐1β, IL‐1α, and INFγ in serum and a significantly younger epigenetic clock as based on DNA methylation levels in blood cells. Overall, our data show that systemic administration of CASIN to reduce Cdc42 activity in aged mice extends murine lifespan.     

Maximizing antibody production in a targeted integration host by optimization of subunit gene dosage and position

Historically, therapeutic protein production in Chinese hamster ovary (CHO) cells has been accomplished by random integration (RI) of expression plasmids into the host cell genome. More recently, the development of targeted integration (TI) host cells has allowed for recombination of plasmid DNA into a predetermined genomic locus, eliminating one contributor to clone-to-clone variability. In this study, a TI host capable of simultaneously integrating two plasmids at the same genomic site was used to assess the effect of antibody heavy chain and light chain gene dosage on antibody productivity. Our results showed that increasing antibody gene copy number can increase specific productivity, but with diminishing returns as more antibody genes are added to the same TI locus. Random integration of additional antibody DNA copies in to a targeted integration cell line showed a further increase in specific productivity, suggesting that targeting additional genomic sites for gene integration may be beneficial. Additionally, the position of antibody genes in the two plasmids was observed to have a strong effect on antibody expression level. These findings shed light on vector design to maximize production of conventional antibodies or tune expression for proper assembly of complex or bispecific antibodies in a TI system.