Human relaxin-2: historical perspectives and role in cancer biology

One of the most recognised and studied family of peptide hormones is the insulin superfamily. Within this family is the relaxin subfamily which comprises seven members: relaxin-1, -2 and -3 and insulin-like peptides 3, 4, 5 and 6. Besides exhibiting sequence similarities, each member exists as an active A-B heterodimer linked by three disulfide bonds. This mini-review is divided into three broad themes: an overview of all insulin superfamily members (including structural similarities); roles of each superfamily member and finally, a focus on the pleiotropic peptide hormone, human relaxin-2. In addition to promoting vasodilatory effects leading to evaluation in Phase III clinical trials for the treatment of acute heart failure, relaxin has recently been shown to be highly expressed by cancer cells, aiding in their proliferation, invasiveness and metastasis. These contrary effects of relaxin are discussed together with current efforts in the development of relaxin antagonists that may possess future therapeutic potential for the treatment of certain cancers.     

Complete observation of all structural,conformational, and fibrillation transitions of monomeric globular proteins at submicellar sodium dodecyl sulfate concentrations

Although considerable information is available regarding protein–sodium dodecyl sulfate (SDS) interactions, it is still unclear as to how much SDS is needed to denature proteins. The role of protein charge and micellar surfactant concentration on amyloid fibrillation is also unclear. This study reports on equilibrium measurements of SDS interaction with six model proteins and analyzes the results to obtain a general understanding of conformational breakdown, reorganization and restructuring of secondary structure, and entry into the amyloid fibrillar state. Significantly, all of these responses are entirely resolved at much lower than the critical micellar concentration (CMC) of SDS. Electrostatic interaction of the dodecyl sulfate anion (DS⁻) with positive surface potential on the protein can completely unfold both secondary and tertiary structures, which is followed by protein chain restructuration to α-helices. All SDS-denatured proteins contain more α-helices than the corresponding native state. SDS interaction stochastically drives proteins to the aggregated fibrillar state.     

Site-specific conjugation of a lanthanide chelator and its effects on the chemical synthesis and receptor binding affinity of human relaxin-2 hormone

Diethylenetriamine pentaacetic acid (DTPA) is a popular chelator agent for enabling the labeling of peptides for their use in structure-activity relationship study and biodistribution analysis. Solid phase peptide synthesis was employed to couple this commercially available chelator at the N-terminus of either the A-chain or B-chain of H2 relaxin. The coupling of the DTPA chelator at the N-terminus of the B-chain and subsequent loading of a lanthanide (europium) ion into the chelator led to a labeled peptide (Eu-DTPA-(B)-H2) in low yield and having very poor water solubility. On the other hand, coupling of the DTPA and loading of Eu at the N-terminus of the A-chain led to a water-soluble peptide (Eu-DTPA-(A)-H2) with a significantly improved final yield. The conjugation of the DTPA chelator at the N-terminus of the A-chain did not have any impact on the secondary structure of the peptide determined by circular dichroism spectroscopy (CD). On the other hand, it was not possible to determine the secondary structure of Eu-DTPA-(B)-H2 because of its insolubility in phosphate buffer. The B-chain labeled peptide Eu-DTPA-(B)-H2 required solubilization in DMSO prior to carrying out binding assays, and showed lower affinity for binding to H2 relaxin receptor, RXFP1, compared to the water-soluble A-chain labeled peptide Eu-DTPA-(A)-H2. The mono-Eu-DTPA labeled A-chain peptide, Eu-DTPA-(A)-H2, thus can be used as a valuable probe to study ligand-receptor interactions of therapeutically important H2 relaxin analogs. Our results show that it is critical to choose an approriate site for incorporating chelators such as DTPA. Otherwise, the bulky size of the chelator, depending on the site of incorporation, can affect yield, solubility, structure and pharmacological profile of the peptide.     

Development of an expressed sequence tag (EST) resource for wheat (Triticum aestivum L.): EST generation, unigene analysis, probe selection and bioinformatics for a 16,000-locus bin-delineated map

This report describes the rationale, approaches, organization, and resource development leading to a large-scale deletion bin map of the hexaploid (2n = 6x = 42) wheat genome (Triticum aestivum L.). Accompanying reports in this issue detail results from chromosome bin-mapping of expressed sequence tags (ESTs) representing genes onto the seven homoeologous chromosome groups and a global analysis of the entire mapped wheat EST data set. Among the resources developed were the first extensive public wheat EST collection (113,220 ESTs). Described are protocols for sequencing, sequence processing, EST nomenclature, and the assembly of ESTs into contigs. These contigs plus singletons (unassembled ESTs) were used for selection of distinct sequence motif unigenes. Selected ESTs were rearrayed, validated by 5′ and 3′ sequencing, and amplified for probing a series of wheat aneuploid and deletion stocks. Images and data for all Southern hybridizations were deposited in databases and were used by the coordinators for each of the seven homoeologous chromosome groups to validate the mapping results. Results from this project have established the foundation for future developments in wheat genomics.     

The Importance of Tryptophan B28 in H2 Relaxin for RXFP2 Binding and Activation

H2 relaxin (relaxin) is a member of the insulin–relaxin superfamily and exhibits several non-reproductive functions in addition to its well-known properties as a pregnancy hormone. Over the years, the therapeutic potential of relaxin has been examined for a number of conditions. It is currently in phase III clinical trials for the treatment of acute heart failure. The 53 amino acid peptide hormone consists of two polypeptide chains (A and B) which are cross-linked by two inter-chains and one intra-A chain disulfide bridge. Although its cognate receptor is relaxin family peptide receptor (RXFP) 1, relaxin is also able to cross-react with RXFP2, for which the native ligand is INSL3. The “RXXXRXXI” motif in the B-chain of H2 relaxin is responsible for primary binding to LRR of the RXFP1 receptor (Büllesbach and Schwabe, J Biol Chem 280:14051–14056, 2005). Previous RXFP2 receptor mutation and molecular modelling studies strongly suggest that, in addition to this motif, the Trp-B28 residue in the B-chain is responsible for H2–RXFP2 interaction. To confirm this finding, here we have mutated H2 relaxin in which Trp-B28 was replaced with alanine. The synthetic relaxin analogue was then tested on cells expressing either RXFP1 or 2 to determine the affinity and potency for the respective receptors. Our results confirm that Trp-B28 in the B-chain is crucial for binding and activating RXFP2, but not for RXFP1.     

Rapid disruption of gap junctional communication and phosphorylation of connexin43 by platelet-derived growth factor in T51B rat liver epithelial cells expressing platelet-derived growth factor receptor

Gap junctional communication (GJC) between contacting cells has been postulated to be involved in the regulation of cell proliferation. This suggestion stems from numerous studies showing modulation of GJC by agents that influence cellular proliferation. Platelet-derived growth factor (PDGF), a strong mitogen, inhibits GJC in many cell types. To understand the molecular nature of the signal transduction pathway responsible for the GJC blockade, T51B rat liver epithelial cells, which lack endogenous PDGF receptor (PDGFr), were infected with a retrovirus containing either wild-type full-length cDNA of human PDGFrβ (Kin⁺) or a mutant PDGFrβ lacking receptor tyrosine kinase activity (Kin⁻). PDGF caused a complete but transient interruption of cell communication in Kin⁺ cells within 15–20 min of addition. This interruption of GJC was not associated with a gross destabilization of gap junction plaques but with the phosphorylation of connexin43 (Cx43), the only known gap junction protein expressed in these cells. These effects were not exhibited in either control T51B cells or in Kin⁻ cells, indicating a requirement of the receptor tyrosine kinase activity. Further examination revealed that the newly phosphorylated Cx43 then undergoes a rapid degradation utilizing the lysosomal pathway resulting in a decreased total Cx43 protein level. The re-establishment of GJC following PDGF treatment was dependent on protein synthesis. This report describes a suitable cell system which is currently being utilized for the characterization of the PDGF signaling pathway responsible for the inhibition of GJC. J. Cell. Physiol. 174:66–77, 1998. © 1998 Wiley-Liss, Inc.     

Optimization of the demineralization process for the extraction of chitin from Omani Portunidae segnis

Chitin is an organic polymer and it is the most frequent marine natural polysaccharide after cellulose. The main natural sources of chitin are exoskeletons of insects, mollusks, the cell walls of certain fungi and crustaceans such as crabs, shrimps and lobsters. The waste of these marine exoskeletons are pollutant for the environment, but these waste raw materials could be useful for production of commercial products like chitin. Chitin is an important raw material used for water treatment, agricultural, biomedical, biotechnological purposes, food and paper industry and cosmetics. Based on the variety of importance, the present targets of this study are to optimize the demineralization process for the removal of calcium and phosphate contents from the waste of Portunidae segnis (P. segnis) by using acid at ambient temperature and to characterize the isolated demineralized sample as well as the percentage of remaining calcium and phosphorus contents by using Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES). The prepared waste carbs coarse powder samples of P. segnis were demineralized with seven different concentrations of hydrochloric acid at ambient temperature for 1 h. All the demineralization samples by the different concentrations were analyzed by using sensitive ICP-OES. The results based on ICP-OES showed that among the seven different concentrations used in the demineralization process for the isolation of chitin, the best was 2 M of HCl concentration for the production of chitin. The results also showed that the optimized concentration 2 M HCl gave the minimum concentration of calcium and phosphorus compared to other concentrations applied in this experiment. In conclusion, the optimized concentration for demineralization process could be used commercially for the isolation or commercial production of chitin for agricultural, biomedical and biotechnological purposes.     

Protective role of tebuconazole and trifloxystrobin in wheat (Triticum aestivum L.) under cadmium stress via enhancement of antioxidant defense and glyoxalase systems

Cadmium (Cd) is a toxic metal and an environmental pollutant that significantly reduces plant growth and productivity. Proper management can ameliorate dysfunction and improve the plant growth and productivity exposed to Cd. Therefore, the present study was conducted to explore the protective role of the fungicides tebuconazole (TEB) and trifloxystrobin (TRI) in helping wheat (Triticum aestivum L. cv. Norin 61) seedlings to tolerate Cd. Five-day-old hydroponically grown seedlings were allowed to mild (0.25 mM CdCl₂) and severe (0.5 mM CdCl₂) Cd stress separately and with the fungicides (2.75 µM TEB + 1.0 µM TRI) for the next four days. Compared to control, the level of H₂O₂ in the seedlings exposed to mild and severe Cd stress alone increased by 81 and 112%, respectively. The accumulation of Cd also increased in the wheat seedlings along with declining mineral nutrients under Cd stress. The protective effect of TEB and TRI was observed with the enhancement of the antioxidant defense and methylglyoxalase systems and reduction in oxidative damage. Applying TEB and TRI reduced MDA (by 9 and 18%), EL (by 21 and 17%), MG (by 12 and 17%), and LOX activity (by 37 and 27%), respectively, relative to Cd stress alone. Cadmium uptake also decreased in the shoots (by 48 and 50%, respectively) and roots (by 23 and 25%, respectively) of the fungicide-treated wheat seedlings under mild and severe Cd stress, relative to stress alone. These results indicate the exogenous application of TEB and TRI is a promising approach to improve Cd tolerance in wheat plants. Further investigation is needed under field conditions and for other crop species to determine the Cd-tolerance induced by TEB and TRI application.     

Development of social sustainability assessment method and a comparative case study on assessing recycled construction materials

Purpose

Sustainability analysis should include the assessment of the environmental, social, and economic impacts throughout the life cycle of a product. However, the social sustainability performance assessment is seldom carried out during materials selection due to its complex nature and the lack of a social life cycle assessment tool. This study presents a single score-based social life cycle assessment methodology, namely social sustainability grading model, for assessing and comparing the social sustainability performance of construction materials using a case study on recycled and natural construction materials.

Methods

The proposed method is developed based on the methodological framework provided by the United Nations Environment Programme/Society of Environmental Toxicology and Chemistry guidelines published in 2009 and the methodological sheets published in 2013, the indicators and sustainability reporting guidelines provided by the Global Reporting Initiatives and ISO 26000 for social responsibility of products, and the indicators provided by the Hong Kong Business Environment Council Limited for construction sustainability. A twofold research approach is proposed in this model: the first one is the qualitative research based on expert interviews to identify, select, and prioritize the relevant subcategories and indicators, and the second one is the operational research based on the case-specific survey to collect the required data. A social sustainability index was proposed for the interpretation of the results effectively. A case study on construction materials was conducted to illustrate the implementation of the method using case-specific first-hand data.

Results and discussion

The major outcome of this study is the systematic development of a social sustainability assessment tool based on the established standards and guidelines. The case study showed that four subcategories are crucial social concerns for construction materials (i.e., health and safety issues of the materials, health and safety of workers, company’s commitment to sustainability, and company’s policies on energy and water consumption). Based on the sustainability index proposed, using recycled aggregates from locally generated waste materials scored higher (about 31–34%) social sustainability than using imported natural aggregates. In addition, recycled aggregates and natural aggregates achieved “sustainable” and “neutral” rating sustainability levels, respectively. However, several subcategories (e.g., health and safety, working hour, forced work, training and social benefits of workers, and quality of the materials and information disclosing to public) are still needed to improve the social sustainability performance of recycled aggregates.

Conclusions

An integrated social life cycle assessment method is presented in this study for assessing the social sustainability of construction materials. In addition, the reported case study in this paper is one of the first attempts for social sustainability assessment of recycled construction materials, and the method can be applied to other recycled materials/products for comparative analysis. However, several critical factors, such as integration in other life cycle methods and software, sensitivity analysis, and more case studies, are still needed for further improvement of the developed method.

     

Length‐weight and length‐length relationships of three small indigenous fishes from the Payra River,southern Bangladesh

Length‐weight relationships (LWRs) and length‐length relationships (LLRs) for three small indigenous fishes (Esomus danrica, Pachypterus atherinoides and Salmostoma bacaila) were reported from the Payra River, southern Bangladesh. Samples were collected using traditional fishing gear including cast net (mesh size ranges from 1.0 to 2.0 cm), seine net (mesh size ranges from 1.5 to 2.5 cm) and square lift net (mesh size ~ 1.0 cm) in August to September 2017. Allometric coefficient (b) values were 2.66 for E. danrica, 3.08 for P. atherinoides and 3.06 for S. bacaila. The LLRs were also highly significant with r² ≥ .956.