TGF-β regulates LARG and GEF-H1 during EMT to affect stiffening response to force and cell invasion

Recent studies implicate a role for cell mechanics in cancer progression. The epithelial-to-mesenchymal transition (EMT) regulates the detachment of cancer cells from the epithelium and facilitates their invasion into stromal tissue. Although classic EMT hallmarks include loss of cell–cell adhesions, morphology changes, and increased invasion capacity, little is known about the associated mechanical changes. Previously, force application on integrins has been shown to initiate cytoskeletal rearrangements that result in increased cell stiffness and a stiffening response. Here we demonstrate that transforming growth factor β (TGF-β)–induced EMT results in decreased stiffness and loss of the normal stiffening response to force applied on integrins. We find that suppression of the RhoA guanine nucleotide exchange factors (GEFs) LARG and GEF-H1 through TGF-β/ALK5–enhanced proteasomal degradation mediates these changes in cell mechanics and affects EMT-associated invasion. Taken together, our results reveal a functional connection between attenuated stiffness and stiffening response and the increased invasion capacity acquired after TGF-β–induced EMT.     

Extensive Miocene speciation in and out of Indochina: The biogeographic history of Typhonium sensu stricto (Araceae) and its implication for the assembly of Indochina flora

The Asian paleotropical flora is characterized by abundance of endemic species, high biodiversity, and complex geological and climatic histories. However, the main driving mechanism underlying such high tropical biodiversity remains unclear. Hence, the present study aimed to investigate the biogeographic origin of the Asian paleotropical flora by tracking the speciation and diversification history of a typical tropical perennial, Typhonium sensu stricto (s.s.) (Araceae), using a time-calibrated whole-plastome phylogeny. In particular, we tested whether the Asian paleotropic region is a macroevolutionary source or sink. We observed that Typhonium s.s. originated in Indochina during the early–middle Miocene epoch, ca. 17.24 Ma (95% highest posterior density [HPD]: 12.83 ̶ 21.99 Ma). Most of the in situ diversification within the genus Typhonium s.s. has been underway since 14.73 Ma, with an accelerated lineage diversification at ca. 15−17 Ma, which may have been triggered by the intensification of the Asian monsoon system around the middle Miocene. Furthermore, the underground tuberous stem of Typhonium s.s. might have played an essential role in the adaptation to the seasonality caused by the monsoon in Indochina. Our results also suggested that peripatric speciation may be important in the diversification of T. trilobatum and T. roxburghii. This study provides a framework for studies in biogeography and evolution of the Asian paleotropical flora.     

The stingless bee honey protects against hydrogen peroxide-induced oxidative damage and lipopolysaccharide-induced inflammation in vitro

Oxidative stress, DNA damage, and unresolved inflammation are the predisposing factors of many chronic and degenerative diseases, including cancer. Stingless bee honey (SBH) is recognized to have high medicinal value by traditional medicine practitioners and has been used to treat various illnesses traditionally. This study aimed to determine the antioxidant, anti-inflammatory, and genoprotective effects of SBH by using in vitro cell culture models. The sugar content, total phenolic content, radical scavenging activity, and ferric reducing antioxidant power (FRAP) of SBH were determined in this study. Then, the protective effect of SBH against hydrogen peroxide (H₂O₂)-induced cell death and DNA damage was studied by using WIL2-NS human lymphoblastoid cell line, while the lipopolysaccharide (LPS)-induced RAW 264.7 murine macrophages cell line was used to study the anti-inflammatory effects of SBH. Results from this present study showed that the major sugar contents of SBH were fructose (19.39 + 0.01%) and glucose (14.03 ± 0.03%). Besides, the total phenolic content, the radical scavenging activity, and the FRAP value of SBH were 15.38 ± 0.02 mg GAE/100 g of honey, 34.04 ± 0.21%, and 206.77 + 1.76 μM AAE/100 g honey respectively. Pretreatment with SBH protected WIL2-NS cells from H₂O₂-induced cell death and DNA damage (p < 0.001). Moreover, SBH was also able to attenuate the production of nitric oxide by inhibiting the expression of inducible nitric oxide synthase in LPS-induced RAW 264.7 cells (p < 0.001). In conclusion, SBH is rich in total phenolic content and possesses strong antioxidant, anti-inflammatory, and genoprotective properties. Our current findings suggest that SBH might be useful in the prevention and treatment of many diseases caused by oxidative stress and inflammation assuming the observed effects are also achievable in vivo.     

Preparation of large temporomandibular joint specimens for pathological study

Ten fresh temporomandibular joint (TMJ) specimens about 5 X 4.5 X 3.5 cm in size were removed at autopsy by 5 cuts according to appropriate anatomical landmarks. After routine formalin fixation, the whole-TMJ specimens were wrapped with a thin layer of self-curing resin and then cut with a low speed bone saw along the parasagittal plane predetermined by x-ray guidance. Each specimen was serially cut into 4 to 5 parallel slices of 3 mm thickness, which were then decalcified with 14% EDTA and embedded in paraffin. Histological sections of 5 microns were stained with hematoxylin and eosin. The procedures were accomplished within 20 to 24 days after autopsy. With this technique, the anatomical interrelationships among the various joint components could be maintained and the macroscopic and microscopic topography of the TMJ could be studied in the desired reference plane. Therefore, the corresponding changes among the joint components in a diseased TMJ could be thoroughly examined. This technique was also applicable for the study of large specimens containing both hard and soft tissues.     

Examination of protein sequence homologies: IV. Twenty-seven bacterial ferredoxins

Summary Sequence homologies of 27 bacterial ferredoxins were examined using a computer program that quantitatively evaluates extent of similarity as a correlation coefficient. The results of a similarity search among the sequences demonstrated that the basal sequence consists of a pair of extremely similar segments of 26 amino acids connected by a three-amino acid group. The segment pairs, which would have arisen from gene duplication, are termed the first and second units. Because of the gene duplication, the connector sequence appears to have been introduced as a structurally important chain reversal. Each of the two units contains four cysteine residues, which are inserted one by one among seven, two, two, three, and eight amino acid alignments, respectively. The bacterial ferredoxins were categorized with regard to basal constitution as follows: group 1, in which both units closely conform to the basal structure; group 2, in which the second unit is modified in a characteristic manner among members; group 3, in which the first unit is modified in a characteristic manner, while the conforming second unit is accompanied by a long accessory sequence; group 4, in which there are modifications before and/or after the units, of which the respective central domains remain nearly intact; and group 5, where only the former of two Fe:S cluster ligation sets of four cysteines is estimated to remain intact, whereas the latter set is extremely modified. It is noteworthy that throughout all bacterial ferredosins, one of two cysteine sets never fails to be completely intact and, moreover, the connector of three amino acids also exists intact. Based on this grouping and on the correspondences among the groups, average correlation coefficients among all members were computed, and the respective evolutionary relationships were examined. The results supported the proposition that transposition had occurred in theAzotobacter-type ferredoxins of group 3.