Wheat TaNACα18 functions as a positive regulator of high-temperature adaptive responses and improves cell defense machinery.

Global wheat production amounted to >780 MMT during 2022-2023 whose market size are valued at >$128 billion. Wheat is highly susceptible to high-temperature stress (HTS) throughout the life cycle and its yield declines 5-7% with the rise in each degree of temperature. Previously, we reported an array of HTS-response markers from a resilient wheat cv. Unnat Halna and described their putative role in heat acclimation. To complement our previous results and identify the key determinants of thermotolerance, here we examined the cytoplasmic proteome of a sensitive cv. PBW343. The HTS-triggered metabolite reprograming highlighted how proteostasis defects influence the formation of an integrated stress-adaptive response. The proteomic analysis identified several promising HTS-responsive proteins, including a NACα18 protein, designated TaNACα18, whose role in thermotolerance remains unknown. Dual localization of TaNACα18 suggests its crucial functions in the cytoplasm and nucleus. The homodimerization of TaNACα18 anticipated its function as a transcriptional coactivator. The complementation of TaNACα18 in yeast and overexpression in wheat demonstrated its role in thermotolerance across the kingdom. Altogether, our results suggest that TaNACα18 imparts tolerance through tight regulation of gene expression, cell wall remodeling and activation of cell defense responses.

A comparative study between bone transport technique using Ilizarov/LRS fixator and induced membrane (Masquelet) technique in management of bone defects in the long bones of lower limb.

Introduction: In earlier times due to difficulty in managing segmental long bone defects, amputation was the preferred treatment. Nonunion with bone loss of long bones is a challenging problem, requiring serious attention. Post-traumatic segmental bone defects can have severe long-term ill impact on patient's lives. Reconstruction is more difficult and functional outcome is usually less satisfactory compared to bony outcome. Distraction osteogenesis and induced membrane technique are the techniques that can be used. Aims and Objectives: To find out and compare clinical, radiological, and functional outcome of bone transport technique and induced membrane technique in management of bone defects in the long bones of lower limb. Materials and Methods: A comparative study was conducted on 24 patients (22 males and 2 females) of lower extremity fractures with bone defect more than 3 cm. Patients were divided into two groups according to the method of reconstruction used, that is, either bone transport technique in 12 patients (group A) or masquelet in the other 12 patients (group B). The mean age of the patients was 44 years in group A and was 38 years in group B. Regular follow-up was done with a mean period of follow up of 18.35 ± 5.58 months in group A and 18.25 ± 3.95 months in group B. Result: In group A (bone transport), 67% showed union, 25% showed union with bone graft and 8% showed delayed union. In group B (masquelet), 75% showed union and 25% showed delayed union. bone transport technique showed excellent results in 58.3% and good in 41.7% while Masquelet technique showed excellent result in 50% and good in 50%. Conclusion: For an orthopaedic surgeon, long bones defects with a substantial loss of bone volume are one of the most challenging bone defects encountered in clinical practice. Induced membrane technique and bone transport both offer successful options for filling of bone defects. Both techniques have its own pros and cons and provide varied option for healing. In our study, both methods have comparable results statistically although induced membrane technique required soft tissue reconstructive procedures.

Comparison of the Efficacy of Platelet-Rich Plasma (PRP) and Local Corticosteroid Injection in Periarthritis Shoulder: A Prospective, Randomized, Open, Blinded End-Point (PROBE) Study.

Background Periarthritis or frozen shoulder, also called adhesive capsulitis, is characterized by stiffness and pain along with gradual loss of active and passive movement in the glenohumeral joint. More than 2-5% of the population suffers from periarthritis with a higher incidence in the age group of 40-60 years. The various treatment modalities used for its management include simple physiotherapy, short-wave therapy, ultrasonic therapy, transcutaneous electrical nerve stimulation, hydrotherapy, analgesics, intra-articular injections, manipulation under general anesthesia (MUA), and surgical management. The application of intra-articular steroid injection has been a common and efficacious option in rapidly diminishing shoulder pain and disability. Some recent studies reported a better outcome using platelet-rich plasma (PRP) injections in frozen shoulder cases. Hence, this randomized controlled trial was conducted to compare the efficacy of intra-articular injections of PRP and triamcinolone in patients of shoulder periarthritis in a population from the eastern region of India Methodology A total of 60 patients with periarthritis shoulder were allocated into two groups after randomization. Group A received 2 mL autologous PRP, and Group B received 2 mL of triamcinolone (40 mg/mL) intra-articular injection. Patients were followed up on the 4th week, 12th week, and 24th week. The assessment of pain and function using the visual analog scale (VAS) score and the Disabilities of Arm, Shoulder, and Hand (DASH) score, respectively, was done at each follow-up. The primary analyses of both primary and secondary outcomes were conducted in the intention-to-treat (ITT) population. SPSS version 24 (IBM Corp., Armonk, NY, USA) was used for data analysis. Results The mean VAS score in the PRP and triamcinolone groups was 14.33 ± 3.79 and 31.63 ± 7.62, respectively (p = 0.0001) after 24 weeks. The mean DASH score in the PRP and triamcinolone groups was 18.08 ± 8.08 and 31.76 ± 3.63, respectively (p = 0.0001), which shows significant improvement in both pain and disability scores in the PRP group after 24 weeks. Conclusions The triamcinolone group showed better short-term outcomes whereas PRP showed better long-term outcomes in reducing pain and disability scores in terms of VAS and DASH scores.

Comparative study to evaluate efficacy of prolotherapy using 25% dextrose and local corticosteroid injection in tennis elbow - A prospective study.

Introduction: Tennis elbow is a common painful condition with a prevalence rate of 1-3%. The condition starts as a micro-tear in extensor carpi radialis brevis and its degeneration because of repetitive overload. Conservative measures are undertaken initially because symptoms in most patients improve with time and rest. Adjunctive treatment includes steroid injection, autologous plasma, exercise, and prolotherapy. Dextrose prolotherapy by virtue of its multi-modal mechanism of action has emerged as a cost-effective treatment option for chronic musculoskeletal and arthritic pain. Materials and Methods: A total of 260 patients were included in the study and divided in two groups for administration of steroids or dextrose prolotherapy. The Visual Analog Scale (VAS) and Mayo Elbow Performance Scale (MEPS) scores were taken for patients at regular follow-up. Results: All patients showed improvement in VAS score and MEPS score following dextrose prolotherapy as well as patients injected with steroids. Conclusion: Both prolotherapy and steroid injection offer adequate pain relief and functional outcome, although prolotherapy has been shown to be superior and given long-lasting results compared to steroid injection in management of tennis elbow.

Comparison of Femoral Neck System Versus Cannulated Cancellous Screws for the Fixation of Femoral Neck Fracture in Young Adults: A Systematic Review and Meta-Analysis.

A femoral neck fracture is a very common injury in the elderly. However, its incidence is low among young adults, usually presenting as an emergency. In young adults, implant choice is one of the important factors. This systematic review aims to analyze the femoral neck system (FNS) versus cannulated cancellous (CC) screw for the fixation of femoral neck fractures in young adults through well-defined objectives. A comprehensive search from the electronic database (PubMed, Google Scholar, Web of Science, Cochrane Library) was conducted from the beginning till February 18, 2022. The data regarding study type, authors, year of publication, country, union time, Harris hip score, intraoperative blood loss, operating time, neck shortening, and hospital stay were extracted from the selected articles and analyzed using RevMan 5.4.1 software. For continuous data, e.g., healing time, intraoperative blood loss, operation time, Harris hip score, neck shortening, and hospital stay, the mean difference (MD), either weighted mean difference (WMD) or standardized mean difference (SMD), with a 95% confidence interval (CI) was recorded. A p-value less than 0.05 was taken as statistically significant. The Newcastle Ottawa scale was used for the risk of bias assessment. Six retrospective cohort studies including 427 patients were selected for the meta-analysis. There was significantly less healing time (WMD= -1.10, 95% CI: -1.73 to -0.47), shorter operation duration (WMD=7.70, 95% CI: -0.06 to 15.46), and better Harris hip score (WMD=4.79, 95% CI: 2.12-7.46) in the FNS than CC screw fixation method. However, intraoperative blood loss was significantly less in the CC screw system (WMD=21.27, 95% CI: 8.20-34.35). There was no significant difference between the two approaches in-hospital stay duration and femoral neck shortening. This can be concluded that FNS is better than CC screw fixation for treating neck of femur fractures in adults on the outcome basis of union time, less operation time, and better Harris hip score (HHS) with significant heterogeneity.

Wheat 2-Cys peroxiredoxin plays a dual role in chlorophyll biosynthesis and adaptation to high temperature.

The molecular mechanism of high-temperature stress (HTS) response, in plants, has so far been investigated using transcriptomics, while the dynamics of HTS-responsive proteome remain unexplored. We examined the adaptive responses of the resilient wheat cultivar 'Unnat Halna' and dissected the HTS-responsive proteome landscape. This led to the identification of 55 HTS-responsive proteins (HRPs), which are predominantly involved in metabolism and defense pathways. Interestingly, HRPs included a 2-cysteine peroxiredoxin (2CP), designated Ta2CP, presumably involved in stress perception and adaptation. Complementation of Ta2CP in yeast and heterologous expression in Arabidopsis demonstrated its role in thermotolerance. Both Ta2CP silencing and overexpression inferred the involvement of Ta2CP in plant growth and chlorophyll biosynthesis. We demonstrated that Ta2CP interacts with protochlorophyllide reductase b, TaPORB. Reduced TaPORB expression was found in Ta2cp-silenced plants, while upregulation was observed in Ta2CP-overexpressed plants. Furthermore, the downregulation of Ta2CP in Taporb-silenced plants and reduction of protochlorophyllide in Ta2cp-silenced plants suggested the key role of Ta2CP in chlorophyll metabolism. Additionally, the transcript levels of AGPase1 and starch were increased in Ta2cp-silenced plants. More significantly, HTS-treated Ta2cp-silenced plants showed adaptive responses despite increased reactive oxygen species and peroxide concentrations, which might help in rapid induction of high-temperature acclimation.

Cultivar-specific high temperature stress responses in bread wheat (Triticum aestivum L.) associated with physicochemical traits and defense pathways.

The increasing global temperature by 1°C is estimated to reduce the harvest index in a crop by 6%, and this would certainly have negative impact on overall plant metabolism. Wheat is one of the most important crops with global annual production of over 600million tonnes. We investigated an array of physicochemical and molecular indexes to unravel differential response of nine commercial wheat cultivars to high temperature stress (HTS). The reduced rate in relative water content, higher membrane stability, slow chlorophyll degradation and increased accumulation of proline and secondary metabolites ingrained higher thermotolerance in cv. Unnat Halna, among others. The altered expression of several stress-responsive genes, particularly the genes associated with photosynthesis, heat shock proteins and antioxidants impinge on the complexity of HTS-induced responses over different genetic backgrounds and connectivity of adaptive mechanisms. This may facilitate the targeted manipulation of metabolic routes in crops for agricultural and industrial exploitation.

Ectopic expression of amaranth seed storage albumin modulates photoassimilate transport and nutrient acquisition in sweetpotato.

Storage proteins in plants, because of high nutrient value, have been a subject of intensive investigation. These proteins are synthesized de novo in the cytoplasm and transported to the storage organelles where they serve as reservoir of energy and supplement of nitrogen during rapid growth and development. Sweetpotato is the seventh most important food crop worldwide, and has a significant contribution to the source of nutrition, albeit with low protein content. To determine the behaviour of seed storage proteins in non-native system, a seed albumin, AmA1, was overexpressed in sweetpotato with an additional aim of improving nutritional quality of tuber proteins. Introduction of AmA1 imparted an increase in protein and amino acid contents as well as the phytophenols. The proteometabolomics analysis revealed a rebalancing of the proteome, with no significant effects on the global metabolome profile of the transgenic tubers. Additionally, the slower degradation of starch and cellulose in transgenic tubers, led to increased post-harvest durability. Present study provides a new insight into the role of a seed storage protein in the modulation of photoassimilate movement and nutrient acquisition.

Comparison of proteomic and metabolomic profiles of two contrasting ecotypes of sweetpotato (Ipomoea batata L.).

UNLABELLED: Sweetpotato has long been acknowledged as a significant contributor of global caloric needs, which continues to be of remarkable economic value. It is an important staple and emergency food in many countries and its annual world production hovers to about 130 million tons. The tubers act as sink and compete for the available photoassimilates eventually leading to the acquisition of nutrients and phytochemicals. Differential display of genes or gene-products, and metabolites causes differences in nutritive value of closely related ecotypes. To better understand the molecular basis for differential nutrient availability and phytochemicals, and exploit the natural genetic variation(s), we aimed at developing proteometabolic profiles of two contrasting ecotypes of sweetpotato. Proteomic analyses led to the identification of 1541 and 1201 proteins in orange fleshed and white fleshed sweetpotato ecotypes, respectively, presumably associated with binding, followed by catalytic, transferase, hydrolase, kinase and transporter activities. Furthermore, metabolome profiling revealed 148 and 126 metabolites in cv. OFSP and WFSP, respectively. This study would provide a basis for future comparative proteometabolomic efforts for sweetpotato, in particular and tuber crops in general. The results would expand our understanding of the proteome as well as metabolome and give new insights into how ecotype-specific traits are developed. BIOLOGICAL SIGNIFICANCE: Sweetpotato, the potato of the tropics, is the seventh most important crop worldwide in terms of production for food and additional industrial resources. Over 95% of the global sweet potato is produced in developing countries where it is considered as emergency food. It is also a vegetable, a snack food and confectionery item in most countries. It greatly contributes as a phytochemical source of nutrition and can produce more edible energy per hectare per day than wheat or rice. The adaptability to a wide range of agroecological conditions with least growth requirements makes it a preferred tuber crop of high commercial significance. Despite its nutritional merits, it has always remained outside the realm of large-scale functional genomics. Therefore, this study was aimed at constructing the proteomics and metabolomics shared resource for sweetpotato. These data are particularly significant, at least partially due to the fact that the currently available information about sweet potato is under-represented.

Comparative analysis of phytochemicals and nutrient availability in two contrasting cultivars of sweet potato (Ipomoea batatas L.).

Sweet potato ranks as the world's seventh most important food crop, and has major contribution to energy and phytochemical source of nutrition. To unravel the molecular basis for differential nutrient availability, and to exploit the natural genetic variation(s) of sweet potato, a series of physiochemical and proteomics experiment was conducted using two contrasting cultivars, an orange-fleshed sweet potato (OFSP) and a white-fleshed sweet potato (WFSP). Phytochemical screening revealed high percentage of carbohydrate, reducing sugar and phenolics in WFSP, whereas OFSP showed increased levels of total protein, flavonoids, anthocyanins, and carotenoids. The rate of starch and cellulose degradation was found to be less in OFSP during storage, indicating tight regulation of gene(s) responsible for starch-degradation. Comparative proteomics displayed a cultivar-dependent expression of proteins along with evolutionarily conserved proteins. These results suggest that cultivar-specific expression of proteins and/or their interacting partners might play a crucial role for nutrient acquisition in sweet potato.

Comparative proteomics reveals a role for seed storage protein AmA1 in cellular growth, development, and nutrient accumulation.

Seed storage proteins are known to be utilized as carbon and nitrogen source for growing seedlings and thus are considered as potential candidates for nutritional improvement. However, their precise function remains unknown. We have earlier shown that ectopic expression of a seed storage protein, AmA1, leads to increase in protein besides high tuber yield in potato. To elucidate the AmA1-regulated molecular mechanism affecting increased protein synthesis, reserve accumulation, and enhanced growth, a comparative proteomics approach has been applied to tuber life-cycle between wild-type and AmA1 potato. The differential display of proteomes revealed 150 AmA1-responsive protein spots (ARPs) that change their intensities more than 2.5-fold. The LC-ESI-MS/MS analyses led to the identification of 80 ARPs presumably associated with cell differentiation, regulating diverse functions, viz., protein biogenesis and storage, bioenergy and metabolism, and cell signaling. Metabolome study indicated up-regulation of amino acids paralleling the proteomics analysis. To validate this, we focused our attention on anatomical study that showed differences in cell size in the cortex, premedullary zone and pith of the tuber, coinciding with AmA1 expression and localization. Further, we interrogated the proteome data using one-way analysis of variance, cluster, and partial correlation analysis that identified two significant protein modules and six small correlation groups centered around isoforms of cysteine protease inhibitor, actin, heat shock cognate protein 83 and 14-3-3, pointing toward AmA1-regulated overlapping processes of protein enhancement and cell growth perhaps through a common mechanism of function. A model network was constructed using the protein data sets, which aim to show how target proteins might work in coordinated fashion and attribute to increased protein synthesis and storage reserve accumulation in AmA1 tubers on one hand and organ development on the other.

Characterisation of the nuclear proteome of a dehydration-sensitive cultivar of chickpea and comparative proteomic analysis with a tolerant cultivar.

Water deficit or dehydration hampers plant growth and development, and shrinks harvest size of major crop species worldwide. Therefore, a better understanding of dehydration response is the key to decipher the regulatory mechanism of better adaptation. In recent years, nuclear proteomics has become an attractive area of research, particularly to study the role of nucleus in stress response. In this study, a proteome of dehydration-sensitive chickpea cultivar (ICCV-2) was generated from nuclei-enriched fractions. The LC-MS/MS analysis led to the identification of 75 differentially expressed proteins presumably associated with different metabolic and regulatory pathways. Nuclear localisation of three candidate proteins was validated by transient expression assay. The ICCV-2 proteome was then compared with that of JG-62, a tolerant cultivar. The differential proteomics and in silico analysis revealed cultivar-specific differential expression of many proteins involved in various cellular functions. The differential tolerance could be attributed to altered expression of many structural proteins and the proteins involved in stress adaptation, notably the ROS catabolising enzymes. Further, a comprehensive comparison on the abiotic stress-responsive nuclear proteome was performed using the datasets published thus far. These findings might expedite the functional determination of the dehydration-responsive proteins and their prioritisation as potential molecular targets for better adaptation.

Next-generation protein-rich potato expressing the seed protein gene AmA1 is a result of proteome rebalancing in transgenic tuber.

Protein deficiency is the most crucial factor that affects physical growth and development and that increases morbidity and mortality especially in developing countries. Efforts have been made to improve protein quality and quantity in crop plants but with limited success. Here, we report the development of transgenic potatoes with enhanced nutritive value by tuber-specific expression of a seed protein, AmA1 (Amaranth Albumin 1), in seven genotypic backgrounds suitable for cultivation in different agro-climatic regions. Analyses of the transgenic tubers revealed up to 60% increase in total protein content. In addition, the concentrations of several essential amino acids were increased significantly in transgenic tubers, which are otherwise limited in potato. Moreover, the transgenics also exhibited enhanced photosynthetic activity with a concomitant increase in total biomass. These results are striking because this genetic manipulation also resulted in a moderate increase in tuber yield. The comparative protein profiling suggests that the proteome rebalancing might cause increased protein content in transgenic tubers. Furthermore, the data on field performance and safety evaluation indicate that the transgenic potatoes are suitable for commercial cultivation. In vitro and in vivo studies on experimental animals demonstrate that the transgenic tubers are also safe for human consumption. Altogether, these results emphasize that the expression of AmA1 is a potential strategy for the nutritional improvement of food crops.