Xylobiose treatment triggers a defense-related response and alters cell wall composition.

Plant cell wall-derived oligosaccharides, i.e., damage-associated molecular patterns (DAMPs), could be generated after pathogen attack or during normal plant development, perceived by cell wall receptors, and can alter immunity and cell wall composition. Therefore, we hypothesised that xylo-oligosaccharides (XOS) could act as an elicitor and trigger immune responses. To test this, we treated Arabidopsis with xylobiose (XB) and investigated different parameters. XB-treatment significantly triggered the generation of reactive oxygen species (ROS), activated MAPK protein phosphorylation, and induced callose deposition. The combination of XB (DAMP) and flg22 a microbe-associated molecular pattern (MAMP) further enhanced ROS response and gene expression of PTI marker genes. RNA sequencing analysis revealed that more genes were differentially regulated after 30 min compared to 24 h XB-treated leaves, which correlated with ROS response. Increased xylosidase activity and soluble xylose level after 30 min and 3 h of XB-treatment were observed which might have weakened the DAMP response. However, an increase in total cell wall sugar and a decrease in uronic acid level was observed at both 30 min and 24 h. Additionally, arabinose, rhamnose, and xylose levels were increased in 30 min, and glucose was increased in 24 h compared to mock-treated leaves. The level of jasmonic acid, abscisic acid, auxin, and cytokinin were also affected after XB treatment. Overall, our data revealed that the shortest XOS can act as a DAMP, which triggers the PTI response and alters cell wall composition and hormone level.

Arabidopsis GELP7 functions as a plasma membrane-localized acetyl xylan esterase, and its overexpression improves saccharification efficiency.

Acetyl substitution on the xylan chain is critical for stable interaction with cellulose and other cell wall polymers in the secondary cell wall. Xylan acetylation pattern is governed by Golgi and extracellular localized acetyl xylan esterase (AXE). We investigated the role of Arabidopsis clade Id from the GDSL esterase/lipase or GELP family in polysaccharide deacetylation. The investigation of the AtGELP7 T-DNA mutant line showed a decrease in stem esterase activity and an increase in stem acetyl content. We further generated overexpressor AtGELP7 transgenic lines, and these lines showed an increase in AXE activity and a decrease in xylan acetylation compared to wild-type plants. Therefore, we have named this enzyme as AtAXE1. The subcellular localization and immunoblot studies showed that the AtAXE1 enzyme is secreted out, associated with the plasma membrane and involved in xylan de-esterification post-synthesis. The cellulose digestibility was improved in AtAXE1 overexpressor lines without pre-treatment, after alkali and xylanases pre-treatment. Furthermore, we have also established that the AtGELP7 gene is upregulated in the overexpressor line of AtMYB46, a secondary cell wall specific transcription factor. This transcriptional regulation can drive AtGELP7 or AtAXE1 to perform de-esterification of xylan in a tissue-specific manner. Overall, these data suggest that AtGELP7 overexpression in Arabidopsis reduces xylan acetylation and improves digestibility properties of polysaccharides of stem lignocellulosic biomass.

First record of off-season flowering in Populus deltoides from India: paradigm of climate change indicator.

Populus deltoides is a fast-growing woody species possessing plethora of industrial applications. This species evolutionarily developed unisexual male and female catkin inflorescence on separate trees. Flowering usually occurs during early spring before the development of foliage, where buds appear near axils or at the extending shoots. In 2019, surveys were undertaken to study the flowering pattern of P. deltoides in the states of Punjab, Haryana, Uttar Pradesh and Uttarakhand in northern India. Interestingly, an anomalous flowering behaviour (appearance of off-season male catkins during autumn, i.e. October) was observed in a plantation trial at Kapurthala, Punjab. The male catkins were 2.7-3.1 ± 0.07 cm long and 0.3-0.5 ± 0.03 cm wide, which is significant for flowering and liberation of pollen grains. Preliminary results suggested that climatic factors, such as episodes of high or low temperature and the precipitation variation forcing the tree species to behave differently. Unearthing the climate-driven off-season flowering in other tree species alluded the stimulation of phytohormones, such as gibberellic and salicylic acid concentrations influencing the flowering time, therefore, needs further investigation in case of P. deltoides. Overall, this work provides early clues of changing climatic scenario altering the flowering pattern of a tropical forestry tree species.

Mitochondrial cytochrome oxidase C subunit III (cox3) gene as a sensitive and specific target for molecular detection of Babesia gibsoni infection in dogs.

A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsonicox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100  µl of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia canis and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.

Cardiac resynchronization therapy device implantation in a patient with persistent left superior vena cava without communicating innominate vein - should we proceed from the same side? - A dilemma revisited.

Persistent left superior vena cava (PLSVC) is an uncommon congenital anomaly. We report a case of implantation of cardiac resynchronization therapy - pacemaker (CRT-P) device in a 38-year-old lady with idiopathic dilated cardiomyopathy. After left axillary vein puncture, we faced an unexpected entry of left subclavian to PLSVC draining into the coronary sinus (CS). The target posterolateral vein which had been identified before, seemed to have an acute angle at its entry into the CS. Hence, at this stage we were in a dilemma, whether to switch to the right side or to continue from the same side. We continued the procedure from the left side and completed it successfully after some manipulation and improvisation.

Downregulation of RWA genes in hybrid aspen affects xylan acetylation and wood saccharification.

High acetylation of angiosperm wood hinders its conversion to sugars by glycoside hydrolases, subsequent ethanol fermentation and (hence) its use for biofuel production. We studied the REDUCED WALL ACETYLATION (RWA) gene family of the hardwood model Populus to evaluate its potential for improving saccharification. The family has two clades, AB and CD, containing two genes each. All four genes are expressed in developing wood but only RWA-A and -B are activated by master switches of the secondary cell wall PtNST1 and PtMYB21. Histochemical analysis of promoter::GUS lines in hybrid aspen (Populus tremula × tremuloides) showed activation of RWA-A and -B promoters in the secondary wall formation zone, while RWA-C and -D promoter activity was diffuse. Ectopic downregulation of either clade reduced wood xylan and xyloglucan acetylation. Suppressing both clades simultaneously using the wood-specific promoter reduced wood acetylation by 25% and decreased acetylation at position 2 of Xylp in the dimethyl sulfoxide-extracted xylan. This did not affect plant growth but decreased xylose and increased glucose contents in the noncellulosic monosaccharide fraction, and increased glucose and xylose yields of wood enzymatic hydrolysis without pretreatment. Both RWA clades regulate wood xylan acetylation in aspen and are promising targets to improve wood saccharification.

Expression of fungal acetyl xylan esterase in Arabidopsis thaliana improves saccharification of stem lignocellulose.

Cell wall hemicelluloses and pectins are O-acetylated at specific positions, but the significance of these substitutions is poorly understood. Using a transgenic approach, we investigated how reducing the extent of O-acetylation in xylan affects cell wall chemistry, plant performance and the recalcitrance of lignocellulose to saccharification. The Aspergillus niger acetyl xylan esterase AnAXE1 was expressed in Arabidopsis under the control of either the constitutively expressed 35S CAMV promoter or a woody-tissue-specific GT43B aspen promoter, and the protein was targeted to the apoplast by its native signal peptide, resulting in elevated acetyl esterase activity in soluble and wall-bound protein extracts and reduced xylan acetylation. No significant alterations in cell wall composition were observed in the transgenic lines, but their xylans were more easily digested by a ß-1,4-endoxylanase, and more readily extracted by hot water, acids or alkali. Enzymatic saccharification of lignocellulose after hot water and alkali pretreatments produced up to 20% more reducing sugars in several lines. Fermentation by Trametes versicolor of tissue hydrolysates from the line with a 30% reduction in acetyl content yielded ~70% more ethanol compared with wild type. Plants expressing 35S:AnAXE1 and pGT43B:AnAXE1 developed normally and showed increased resistance to the biotrophic pathogen Hyaloperonospora arabidopsidis, probably due to constitutive activation of defence pathways. However, unintended changes in xyloglucan and pectin acetylation were only observed in 35S:AnAXE1-expressing plants. This study demonstrates that postsynthetic xylan deacetylation in woody tissues is a promising strategy for optimizing lignocellulosic biomass for biofuel production.

Squamous cell carcinoma of tongue and buccal mucosa: clinico-pathologically different entities.

The objective of this study was to examine the clinical and pathological features of squamous cell carcinoma of the Tongue and Buccal Mucosa and understand their differences. This is a retrospective analysis of prospectively collected data of 735 patients with squamous cell carcinoma of the tongue and 665 cases of carcinoma of the buccal mucosa treated by surgery at our hospital. Statistical analysis was done to examine clinical and pathological differences between carcinoma of the tongue and the buccal mucosa with regards to age, gender, clinical T stage/N stage, pathological T stage/N stage, overall stage, grade, thickness, perinodal extension (PNE), lymphovascular emboli (LVE) and perineural invasion (PNI). Statistically significant differences were found for factors like age (p < 0.001), gender (p < 0.001), clinical T staging (p < 0.001) and pathological stage (p < 0.001), grade of tumor (p < 0.001) and perineural invasion (p < 0.001) between carcinoma of the tongue and the buccal mucosa. Forty-eight percent patients in either subsite had pathologically proven node negative necks (pN0, p = 0.88). Multivariate analysis for occult nodal metastases revealed that predictive factors were different for the two subsites. There are significant differences between cancers of the tongue and buccal mucosa for various clinical and pathological factors. This may be a reflection of the underlying differences in their causation and pathophysiology. Squamous cell carcinoma in these two subsites should therefore be regarded as clinico-pathologically distinct entities.

Populus GT43 family members group into distinct sets required for primary and secondary wall xylan biosynthesis and include useful promoters for wood modification.

The plant GT43 protein family includes xylosyltransferases that are known to be required for xylan backbone biosynthesis, but have incompletely understood specificities. RT-qPCR and histochemical (GUS) analyses of expression patterns of GT43 members in hybrid aspen, reported here, revealed that three clades of the family have markedly differing specificity towards secondary wall-forming cells (wood and extraxylary fibres). Intriguingly, GT43A and B genes (corresponding to the Arabidopsis IRX9 clade) showed higher specificity for secondary-walled cells than GT43C and D genes (IRX14 clade), although both IRX9 and IRX14 are required for xylosyltransferase activity. The remaining genes, GT43E, F and G (IRX9-L clade), showed broad expression patterns. Transient transactivation analyses of GT43A and B reporters demonstrated that they are activated by PtxtMYB021 and PNAC085 (master secondary wall switches), mediated in PtxtMYB021 activation by an AC element. The high observed secondary cell wall specificity of GT43B expression prompted tests of the efficiency of its promoter (pGT43B), relative to the CaMV 35S (35S) promoter, for overexpressing a xylan acetyl esterase (CE5) or downregulating REDUCED WALL ACETYLATION (RWA) family genes and thus engineering wood acetylation. CE5 expression was weaker when driven by pGT43B, but it reduced wood acetyl content substantially more efficiently than the 35S promoter. RNAi silencing of the RWA family, which was ineffective using 35S, was achieved when using GT43B promoter. These results show the utility of the GT43B promoter for genetically engineering properties of wood and fibres.

Reduced Wall Acetylation proteins play vital and distinct roles in cell wall O-acetylation in Arabidopsis.

The Reduced Wall Acetylation (RWA) proteins are involved in cell wall acetylation in plants. Previously, we described a single mutant, rwa2, which has about 20% lower level of O-acetylation in leaf cell walls and no obvious growth or developmental phenotype. In this study, we generated double, triple, and quadruple loss-of-function mutants of all four members of the RWA family in Arabidopsis (Arabidopsis thaliana). In contrast to rwa2, the triple and quadruple rwa mutants display severe growth phenotypes revealing the importance of wall acetylation for plant growth and development. The quadruple rwa mutant can be completely complemented with the RWA2 protein expressed under 35S promoter, indicating the functional redundancy of the RWA proteins. Nevertheless, the degree of acetylation of xylan, (gluco)mannan, and xyloglucan as well as overall cell wall acetylation is affected differently in different combinations of triple mutants, suggesting their diversity in substrate preference. The overall degree of wall acetylation in the rwa quadruple mutant was reduced by 63% compared with the wild type, and histochemical analysis of the rwa quadruple mutant stem indicates defects in cell differentiation of cell types with secondary cell walls.

A Prospective Study on Fixation of Syndesmotic Ankle Injury: Tight Rope Versus Screw Fixation.

Background Syndesmotic injury can result in significant instability and long-term complications if not treated correctly. Traditional management has involved transyndesmotic screw fixation, but a newer technique, the tight rope system, has been developed to mitigate some of the issues related to screw fixation, such as hardware discomfort and the necessity for hardware removal. Methods In this randomized, prospective study, 32 patients with ankle injuries requiring syndesmotic fixation were equally divided into two groups: one receiving the tight rope system (n=16) and the other undergoing screw fixation (n=16). The patients were monitored for six months following surgery. The study measured outcomes such as time to weight-bearing, range of motion, pain levels, functional outcomes using the American Orthopaedic Foot & Ankle Society (AOFAS) Ankle-Hindfoot Scale, and complication rates. Results Both groups had comparable demographic and injury profiles. The tight rope group achieved weight-bearing significantly earlier (6.19 ± 0.9 weeks vs. 7.13 ± 0.95 weeks, p=0.008) and had better functional outcomes at six months (87.5% excellent AOFAS scores vs. 37.5%, p=0.003) compared to the screw fixation group. The range of motion and pain scores were similar between the groups. Different complications were observed: screw breakage was more common in the screw fixation group, while the tight rope group experienced more laxity. Overall complication rates were similar. Conclusion Both techniques were effective in reducing pain and maintaining range of motion. However, the tight rope system allowed for earlier weight-bearing and better functional outcomes at six months. These results indicate that the tight rope system may provide certain advantages in treating syndesmotic injuries, although the choice of technique should be tailored to the specific injury and patient factors.

Combining Fourier-transform infrared spectroscopy and multivariate analysis for chemotyping of cell wall composition in Mungbean (Vigna radiata (L.) Wizcek).

BACKGROUND: Dissection of complex plant cell wall structures demands a sensitive and quantitative method. FTIR is used regularly as a screening method to identify specific linkages in cell walls. However, quantification and assigning spectral bands to particular cell wall components is still a major challenge, specifically in crop species. In this study, we addressed these challenges using ATR-FTIR spectroscopy as it is a high throughput, cost-effective and non-destructive approach to understand the plant cell wall composition. This method was validated by analysing different varieties of mungbean which is one of the most important legume crops grown widely in Asia. RESULTS: Using standards and extraction of a specific component of cell wall components, we assigned 1050-1060 cm-1 and 1390-1420 cm-1 wavenumbers that can be widely used to quantify cellulose and lignin, respectively, in Arabidopsis, Populus, rice and mungbean. Also, using KBr as a diluent, we established a method that can relatively quantify the cellulose and lignin composition among different tissue types of the above species. We further used this method to quantify cellulose and lignin in field-grown mungbean genotypes. The ATR-FTIR-based study revealed the cellulose content variation ranges from 27.9% to 52.3%, and the lignin content variation ranges from 13.7% to 31.6% in mungbean genotypes. CONCLUSION: Multivariate analysis of FT-IR data revealed differences in total cell wall (600-2000 cm-1), cellulose (1000-1100 cm-1) and lignin (1390-1420 cm-1) among leaf and stem of four plant species. Overall, our data suggested that ATR-FTIR can be used for the relative quantification of lignin and cellulose in different plant species. This method was successfully applied for rapid screening of cell wall composition in mungbean stem, and similarly, it can be used for screening other crops or tree species.

Modifying lignin composition and xylan O-acetylation induces changes in cell wall composition, extractability, and digestibility.

BACKGROUND: Lignin and xylan are important determinants of cell wall structure and lignocellulosic biomass digestibility. Genetic manipulations that individually modify either lignin or xylan structure improve polysaccharide digestibility. However, the effects of their simultaneous modifications have not been explored in a similar context. Here, both individual and combinatorial modification in xylan and lignin was studied by analysing the effect on plant cell wall properties, biotic stress responses and integrity sensing. RESULTS: Arabidopsis plant co-harbouring mutation in FERULATE 5-HYDROXYLASE (F5H) and overexpressing Aspergillus niger acetyl xylan esterase (35S:AnAXE1) were generated and displayed normal growth attributes with intact xylem architecture. This fah1-2/35S:AnAXE1 cross was named as hyper G lignin and hypoacetylated (HrGHypAc) line. The HrGHypAc plants showed increased crystalline cellulose content with enhanced digestibility after chemical and enzymatic pre-treatment. Moreover, both parents and HrGHypAc without and after pre-treating with glucuronyl esterase and alpha glucuronidase exhibited an increase in xylose release after xylanase digestion as compared to wild type. The de-pectinated fraction in HrGHypAc displayed elevated levels of xylan and cellulose. Furthermore, the transcriptomic analysis revealed differential expression in cell wall biosynthetic, transcription factors and wall-associated kinases genes implying the role of lignin and xylan modification on cellular regulatory processes. CONCLUSIONS: Simultaneous modification in xylan and lignin enhances cellulose content with improved saccharification efficiency. These modifications loosen cell wall complexity and hence resulted in enhanced xylose and xylobiose release with or without pretreatment after xylanase digestion in both parent and HrGHypAc. This study also revealed that the disruption of xylan and lignin structure is possible without compromising either growth and development or defense responses against Pseudomonas syringae infection.

Combined Deltoid and Spring Ligament Reconstruction Using the Quadrangular Construct: Our Experience and Review of the Literature.

Background A combined reconstruction of chronic deltoid and spring ligament insufficiency is uncommon. Our study aims to share our experience in treating post-traumatic, chronic deltoid, and spring ligament insufficiency using the "quadrangular construct" technique. Material and methods Five patients who had post-traumatic combined deltoid and spring ligament insufficiency were included in the study. All patients reported a "giving-way" sensation. Preoperatively, each patient underwent weight-bearing radiographs of the ankle and foot. The talo-first metatarsal angle and hindfoot alignment angle were noted. The superficial deltoid ligament was repaired using a suture anchor augmented with Internal BraceTM (Arthrex, Naples, USA) FiberTape® to form a quadrangular construct that anatomically mimics various components of the deltoid-spring ligament complex. Due to the associated excessive heel valgus, three patients also underwent medial displacement calcaneum osteotomy. Additionally, one patient required lateral ligament repair, and another patient required syndesmotic stabilization. The American Orthopaedic Foot and Ankle Society (AOFAS) hindfoot score was used to evaluate preoperative and postoperative ankle function. Results All five patients were followed up for a mean of 20 months (range: 12-24 months). The mean preoperative talo-first metatarsal angle improved from 8.46 degrees to 4.84 degrees. The preoperative mean hindfoot alignment angle was reduced from 10.9 to 5.76 degrees postoperatively. One patient had irritation due to the anchor, which needed removal after one year. Postoperatively, no patients re-experienced the feeling of "giving way". The AOFAS scores postoperatively showed two patients as excellent, two as good, and one as fair. All the patients returned to their pre-injury work. Conclusion  We have developed a technique for combined deltoid and spring ligament reconstruction using a quadrangular construct. This technique helps to restore anatomical stability, is safe, easily reproducible, and has shown positive short-term results in follow-up. The level of evidence is one of the methods used to categorize the quality and reliability of research, and our study falls under the category of level IV evidence.

Assessing effectiveness of braille and audio-tactile performance technique in improving oral hygiene status of young adults with visual impairment.

INTRODUCTION: Young adults with Visual impairment (VI) challenge the dentists' skills and knowledge due to a higher risk of developing oral diseases due to difficulty in attaining good oral hygiene (OH). AIMS: To assess the effectiveness of ATP (Audio-Tactile Performance) technique and braille versus braille alone in improving OH status of young adults with VI. METHODS AND MATERIAL: A parallel arm, randomized controlled trial was conducted among 70 young adults with VI. Participants were randomly allocated to either test group (braille combined with ATP) or control group (braille alone). A pre-validated braille questionnaire was used to obtain the baseline data followed by clinical examination. OH status assessment was recorded through Gingival Index (GI) and Plaque Index (PI) proforma followed by a thorough ultrasonic oral prophylaxis. Periodic reinforcement was carried out on the 7th day, after 1 month and 3 months. The outcomes were assessed at the end of 3rd and 6th month. RESULTS: There was an increase in the knowledge scores after 3 and 6 months and attitude scores, GI scores and PI scores after 6 months in the test group compared to the control group and this difference was found to be statistically significant. CONCLUSIONS: The findings of this study showed that a combination of ATP and braille proved to be more effective than braille alone in improving the knowledge and OH status of young adults with VI.

Morpho-biochemical characterization of a RIL population for seed parameters and identification of candidate genes regulating seed size trait in lentil (Lens culinaris Medik.).

The seed size and shape in lentil (Lens culinaris Medik.) are important quality traits as these influences the milled grain yield, cooking time, and market class of the grains. Linkage analysis was done for seed size in a RIL (F5:6) population derived by crossing L830 (20.9 g/1000 seeds) with L4602 (42.13 g/1000 seeds) which consisted of 188 lines (15.0 to 40.5 g/1000 seeds). Parental polymorphism survey using 394 SSRs identified 31 polymorphic primers, which were used for the bulked segregant analysis (BSA). Marker PBALC449 differentiated the parents and small seed size bulk only, whereas large seeded bulk or the individual plants constituting the large-seeded bulk could not be differentiated. Single plant analysis identified only six recombinant and 13 heterozygotes, of 93 small-seeded RILs (<24.0 g/1000 seed). This clearly showed that the small seed size trait is very strongly regulated by the locus near PBLAC449; whereas, large seed size trait seems governed by more than one locus. The PCR amplified products from the PBLAC449 marker (149bp from L4602 and 131bp from L830) were cloned, sequenced and BLAST searched using the lentil reference genome and was found amplified from chromosome 03. Afterward, the nearby region on chromosome 3 was searched, and a few candidate genes like ubiquitin carboxyl-terminal hydrolase, E3 ubiquitin ligase, TIFY-like protein, and hexosyltransferase having a role in seed size determination were identified. Validation study in another RIL mapping population which is differing for seed size, showed a number of SNPs and InDels among these genes when studied using whole genome resequencing (WGRS) approach. Biochemical parameters like cellulose, lignin, and xylose content showed no significant differences between parents and the extreme RILs, at maturity. Various seed morphological traits like area, length, width, compactness, volume, perimeter, etc., when measured using VideometerLab 4.0 showed significant differences for the parents and RILs. The results have ultimately helped in better understanding the region regulating the seed size trait in genomically less explored crops like lentils.

Surgeon administered direct adductor canal block is as good as ultrasound guided adductor canal block in pain management in knee replacements- A retrospective case-control study.

Aim: The aim of this study is to assess the feasibility of the DACB in a clinical setting and compare the efficacy of postoperative pain relief after TKR in the patients administered DACB versus USG guided ACB. Also to see efficacy and safety of USACB in patients operated with medial parapatellar and subvastus approach. Material and methods: 250 consecutive patients operated with TKR between Jan 2019 to March 2022 were included. Group A included patients operated with medial parapatellar approach and received USACB, Group B included patients operated with medial parapatellar approach and received DACB while Group C included patients operated with subvastus approach and received USACB. VAS scores between three groups were compared at 12 and 24 h. All three groups of patients were otherwise treated identically in the hospital. Results: The mean age and BMI was not statistically significant between the three groups. The mean VAS pain score at rest at 12 h was 3.06 ± 1.49 (Group A) vs 1.58 ± 1.19 (Group B) [p < 0.0001] and 3.06 ± 1.49 (Group A) vs 1.88 ± 1.18 (Group C) [p < 0.0001]; and at 24 h was 1.88 ± 1.31 (Group A) vs 2.39 ± 1.27 (Group B) [p = 0.023] and 1.88 ± 1.31 (Group A) vs 2.19 ± 1.29 (Group C) [p = 0.16]. The mean theatre time was 151.9 ± 11.37 min (Group A) vs 141.02 ± 19.46 min (Group B) (p = 0.0003) and 151.9 ± 11.37 min (Group A) vs 150.4 ± 28.74 min (Group C) (p = 0.72). Hospital stay was 3.82 ± 0.80 (Group A) vs 4.0 ± 1.09 (Group B) [p = 0.30] and 3.82 ± 0.80 (Group A) vs 2.7 ± 0.69 (Group C) [p < 0.0001]. Group B and Group C had one complication each. Conclusion: USG ACB irrespective of approach used remains the gold standard in providing consistent pain relief and thereby facilitating early discharge. However, increased operating room turnover time and repeated top-ups remain a disadvantage. Both the quantum of pain relief and the potential downsides remained the same irrespective of the surgical approach used and whether or not steroid was added to the cocktail used for infiltration. On the other hand, DACB provides a short lasting (24 h) adequate pain relief after TKR with similar low complication rates. The technique of DACB may have a potential for a wider use especially in centres where outpatient arthroplasties are performed, if newer longer acting anaesthetic/analgesic combinations are devised.

Comparative transcriptome analysis, unfolding the pathways regulating the seed-size trait in cultivated lentil (Lens culinaris Medik.).

Market class, cooking time, quality, and milled grain yield are largely influenced by the seed size and shape of the lentil (Lens culinaris Medik.); thus, they are considered to be important quality traits. To unfold the pathways regulating seed size in lentils, a transcriptomic approach was performed using large-seeded (L4602) and small-seeded (L830) genotypes. The study has generated nearly 375 million high-quality reads, of which 98.70% were properly aligned to the reference genome. Among biological replicates, very high similarity in fragments per kilobase of exon per million mapped fragments values (R > 0.9) showed the consistency of RNA-seq results. Various differentially expressed genes associated mainly with the hormone signaling and cell division pathways, transcription factors, kinases, etc. were identified as having a role in cell expansion and seed growth. A total of 106,996 unigenes were used for differential expression (DE) analysis. String analysis identified various modules having certain key proteins like Ser/Thr protein kinase, seed storage protein, DNA-binding protein, microtubule-associated protein, etc. In addition, some growth and cell division-related micro-RNAs like miR3457 (cell wall formation), miR1440 (cell proliferation and cell cycles), and miR1533 (biosynthesis of plant hormones) were identified as having a role in seed size determination. Using RNA-seq data, 5254 EST-SSR primers were generated as a source for future studies aiming for the identification of linked markers. In silico validation using Genevestigator® was done for the Ser/Thr protein kinase, ethylene response factor, and Myb transcription factor genes. It is of interest that the xyloglucan endotransglucosylase gene was found differentially regulated, suggesting their role during seed development; however, at maturity, no significant differences were recorded for various cell wall parameters including cellulose, lignin, and xylose content. This is the first report on lentils that has unfolded the key seed size regulating pathways and unveiled a theoretical way for the development of lentil genotypes having customized seed sizes.

Comparative Study of Pinless Navigation System versus Conventional Instrumentation in Total Knee Arthroplasty.

BACKGROUD: Optimal placement of the components and achieving a neutral mechanical axis are the main goals of total knee arthroplasty (TKA). Different computerised navigation systems are presently used for these purposes. This aim of this study was to compare the pinless navigation (PNA) TKA performed using iAssist with the conventional instrumented (CIN) TKA in terms of functional and radiological outcomes. METHODS: A total of 100 knees operated for TKA by a single surgeon were studied retrospectively for a period of 2 years. Weight-bearing postoperative radiographs of the knees along with scanograms of the lower limbs were used for measurements of component positioning, mechanical axis alignment, and number of outliers. Oxford knee scoring was used for functional analysis. RESULTS: No statistically significant difference was seen in the mean mechanical axis alignment (hip-knee-ankle angle), coronal alignment (α and ß angles) and sagittal alignment (γ and δ angles) of the femoral and tibial components between the two groups. Though the percentage of outliers for mechanical axis alignment was lower in the PNA-TKA group than in the CIN-TKA group, the difference was not statistically significant (p = 0.73). The number of outliers for the femoral and tibial component positioning in coronal and sagittal planes was not statistically significantly different between the two groups. No statistically significant difference (p = 0.68) was noted between the two groups with respect to the Oxford Knee Score. The mean surgical time was greater in the PNA-TKA group by 11 minutes, which was statistically significantly longer (p = 0.018). Complications were seen in 6.89% of the cases in the CIN-TKA group, while none in the PNA-TKA group. CONCLUSIONS: The accurate mechanical axis alignment and component positioning can be achieved with the conventional instrumentation, so the use of PNA system, which adds to the surgical cost, is questionable. Also, equally good short-term functional outcome can be achieved with the conventional instrumentation. The surgeon must be accustomed with the instrumentation of the PNA system, or it adds to the surgical time.

Roof arc width: The novel calculation method for calculation of patient specific roof arc width in acetabular fractures.

PURPOSE: Roof arc angle (RAA) is determined by measuring angle between a vertical line drawn from center of the acetabulum towards the acetabular dome and a second line drawn from center of acetabulum to the fracture through the acetabulum. Joel and Matta demonstrated that when roof arc angle was less than 45° on Pelvic AP and Judet's views, the fracture line is considered to be passing through the weight-bearing dome and require surgical fixation. The main purpose of the study is to calculate patient-specific angle and width for the better evaluation and management of acetabular fractures. METHODS: Radiographs of normal hips were retrieved from electronic data and parameters were calculated. Two observers calculated the parameters at two different intervals. Pearson correlation formula was used to find a correlation between groups. RESULTS: Fifty radiographs of 28 patients were reviewed. The mean age of patients was 75.58 years ±13.28. The radius of the acetabulum, the radiographic measurement of sector width for 45° angulation at the roof, and the mathematical calculation for roof arc for 45° angle had significant positive correlation for both observers at two different occasions. CONCLUSIONS: The measured roof arc width ranges from a minimum of 16.20 mm-31.50 mm and the calculated arc width for a 45-degree angle varies from a minimum of 15 mm-25.56 mm. These values are higher than the described values of 10 mm equals to 45 degrees. Hence, the values measured in this study should be considered for decision making in the management of acetabular fractures.