Sterol partitioning by HMGR and DXR for routing intermediates toward withanolide biosynthesis.

Withanolides biosynthesis in the plant Withania somnifera (L.) Dunal is hypothesized to be diverged from sterol pathway at the level of 24-methylene cholesterol. The conversion and translocation of intermediates for sterols and withanolides are yet to be characterized in this plant. To understand the influence of mevalonate (MVA) and 2-C-methyl-d-erythritol-4-phosphate (MEP) pathways on sterols and withanolides biosynthesis in planta, we overexpressed the WsHMGR2 and WsDXR2 in tobacco, analyzed the effect of transient suppression through RNAi, inhibited MVA and MEP pathways and fed the leaf tissue with different sterols. Overexpression of WsHMGR2 increased cycloartenol, sitosterol, stigmasterol and campesterol compared to WsDXR2 transgene lines. Increase in cholesterol was, however, marginally higher in WsDXR2 transgenic lines. This was further validated through transient suppression analysis, and pathway inhibition where cholesterol reduction was found higher due to WsDXR2 suppression and all other sterols were affected predominantly by WsHMGR2 suppression in leaf. The transcript abundance and enzyme analysis data also correlate with sterol accumulation. Cholesterol feeding did not increase the withanolide content compared to cycloartenol, sitosterol, stigmasterol and campesterol. Hence, a preferential translocation of carbon from MVA and MEP pathways was found differentiating the sterols types. Overall results suggested that MVA pathway was predominant in contributing intermediates for withanolides synthesis mainly through the campesterol/stigmasterol route in planta.

Simultaneous Intrusion and Retraction of Maxillary Anterior Teeth in Class II Malocclusion Using the Kalra Simultaneous Intrusion and Retraction (K-SIR) Loop: A Case Report.

The straight-wire device offers the best finishing potential and control. This case study focuses on the treatment of severe deep bite and Class II malocclusion involving first premolar extraction of the upper arch using a Kalra Simultaneous Intrusion and Retraction loop. Using minimal force and creating enough space for anterior teeth to retract while maintaining the Class II molar relationship was the aim of the therapy. Due to the unsightly excessive maxillary incisor showing at rest, the decision was made to intrude anterior teeth to treat a deep overbite. Good and consistent changes occurred post-treatment.

Strategic Intervention: Ectopic Premolar Management for Preventing Molar-Premolar Transposition in Class III Malocclusion.

Class III malocclusion often leads to the ectopic eruption of premolars in the upper arch, posing unique challenges for orthodontic practitioners. This case report explores the clinical implications of ectopic maxillary premolars in a Class III malocclusion patient, emphasizing the importance of early intervention and comprehensive treatment strategies. Factors contributing to ectopic eruption, including genetic predisposition and anatomical variations, are discussed, guiding orthodontists in effective diagnosis and management. A 14-year-old patient with Class III malocclusion, anterior crossbite, and ectopic maxillary premolars sought orthodontic consultation. The treatment plan involved extracting the deciduous second molar, realigning the ectopic premolar, and addressing arch length discrepancies. Utilizing fixed orthodontic appliances and strategic force application, the patient achieved Class I molar and canine relationships, resolving the ectopic premolar alignment within 10 months. Ectopic eruption of maxillary premolars, especially in Class III malocclusion, is a critical concern for oral health. Genetic predisposition, arch crowding, and developmental disturbances contribute to this condition. Early intervention, as demonstrated in this case, plays a pivotal role in restoring dentoskeletal harmony. The study underscores the need for a multidisciplinary approach, combining orthodontic, surgical, and restorative interventions for optimal outcomes. Thus, this case report highlights the successful management of ectopic maxillary premolars in a Class III malocclusion patient through strategic orthodontic intervention. Understanding the etiological factors and employing a comprehensive treatment approach facilitate timely diagnosis and prevent complications. Orthodontists must navigate the complexities of ectopic eruption, considering occlusal effects and collaborating with other specialists for holistic patient care.

Unveiling the Journey: A Case Report of Managing an Impacted Central Incisor.

Although impaction of the maxillary permanent central incisor is uncommon in dentistry due to its significance to facial aesthetics which are challenging to treat. To abstain from the consequences related to aesthetic and functional occlusion, early detection of an impacted central incisor is imperative. This case report describes a male patient, aged 22 years, who had an impacted central incisor tooth in the maxillary anterior region. A surgery was performed to remove the impacted supernumerary tooth that was preventing the eruption of the central incisor. Using a combination of surgical exposure and orthodontic force, the impacted right maxillary central incisor was relocated to its proper occlusion in the dental arch.

Mini-Screw-Assisted Orthodontic Retraction of Class I Malocclusion in a Bimaxillary Protrusion Patient Using Sliding Mechanics: A Case Report.

A condition known as bimaxillary protrusion occurs when the front teeth protrude due to the forward positioning of the lower and upper jaws. Temporary anchorage devices (TADs) are utilized to provide anchorage and facilitate the controlled retraction of maxillary and mandibular protruding teeth, helping to correct the patient's bite and facial aesthetics. A 27-year-old female with bimaxillary protrusion reported to the Department of Orthodontics. On examination, the facial profile of the patient was convex. The clinical FMA was high. With a deep mentolabial sulcus and an acute nasolabial angle, lips were potentially competent. An intraoral examination revealed proclined incisors with spacing in the maxillary arch and proclined anterior teeth in the mandibular arch. Space closure was done using sliding mechanics with direct anchorage from a mini-screw after the extraction of all four first premolars. There was a significant improvement in the patient's profile posttreatment.

Morphometric Analysis of Radix Entomolaris: Implications for Endodontic Access and Treatment.

Radix entomolaris (RE) is an anatomical variation that involves the existence of an extra root in lower molars, particularly the first molar. This variant, although less common, has significant clinical implications in endodontic treatment and dental surgeries. Appropriate detection of radix entomolaris is crucial for treatment planning and prognosis. Various diagnostic aids like radiography, cone-beam computed tomography, and intraoperative exploration aid in identifying this anatomical variant.

Overview of Interim and Temporary Restorations of Teeth During Endodontic Treatment.

Root canal treatment of vital, non-infected teeth can often be completed in a single visit, negating the necessity for dressing and provisionalization. Conversely, cases involving infected canals typically demand multiple visits, during which antibacterial medicaments are applied, making effective provisionalization crucial for varying durations. The key components of a successful root canal treatment include adequate canal shape to promote efficient obturation, thorough chemical and mechanical debridement, and complete removal of pulp tissue remnants and bacteria. The primary cause of pain following the initiation of endodontic treatments is often attributed to inadequate debridement or incomplete removal of the pulp tissue, closely followed by insufficient temporary restorations. This review aims to comprehensively overview provisionalization materials used during and immediately after endodontic procedures.

A transcriptomic approach for exploring the molecular basis for dosha-balancing property-based classification of plants in Ayurveda.

In Ayurveda, a healthy body is defined by a balance among the three doshas (Vata, Pitta, Kapha) and ailments result due to imbalances among them. It prescribes specific plant parts/tissues collected in a season-specific manner for curing dosha-related imbalances but the plants prescribed for treating a particular dosha imbalance belong to taxonomically diverse families and often contain similar classes of phytomolecules, making it difficult to provide a phytochemical validation for any similarity that might exist among them. This exploratory study hypothesised that plants of the same dosha-curing group may have similarity at the transcript level. For proving/disproving the hypothesis, cDNA-AFLP and specific expression subset analysis (SESA) were carried out on the Ayurveda-defined active tissues of four representative plants each of the three dosha-balancing groups. cDNA-AFLP analyses indicated that even though the plants belonging to a particular dosha-group may widely differ at the transcript level, there is a small fraction of transcripts that is monomorphic among their active tissues. SESA (Tester-active tissue cDNA; Driver-cDNA from other major tissue[s]) generated 803 subtractive ESTs from the twelve plants that yielded 150 unigenes upon assembly (of ESTs from each plant separately). Cross-plant EST assembly for plants in the same dosha group also corroborated the results. Although a distinct pattern of transcripts was not observed across all the plants in a particular dosha group, some commonalities were obtained that need further characterization towards searching for the hitherto elusive similarity among plants of the same group.

Impact of essential metals on insulin sensitivity and fetuin in obesity-related type 2 diabetes pathogenesis.

Purpose: Essential metals may be crucial in obesity and type 2 diabetes (T2DM); diabesity pathogenesis and consequences. This study aimed to determine the metal levels in obese and non-obese patients with and without T2DM and their relationships with fetuin-A(Fet-A) levels, insulin sensitivity, and insulin resistance. Methods: A total of 314 participants were enrolled, with 160 newly diagnosed T2DM patients and 154 non-T2DM subjects categorized into diabetic obese (n = 57), diabetic non-obese (n = 103), non-diabetic obese (n = 48), and non-diabetic non-obese (n = 106) subgroups. Fet-A, insulin sensitivity (QUCKI)/resistance (HOMA-IR), fasting glucose, and body mass index (BMI) were assessed. The essential metals were measured using inductively coupled plasma mass spectroscopy (ICP-MS). Results: Fet-A levels were 3-fold higher (1391.4 ± 839.8 ng/ml) in T2DM patients than in non-T2DM (2165.6 ± 651.9 vs. 424.3 ± 219.1 ng/ml, p < 0.0001). Fet-A levels were 2.3-fold higher in the diabetic obese group than in the diabetic non-obese group (p < 0.0001). Fet-A levels were 2.0-fold higher in the diabetic non-obese group than in the non-diabetic obese group (p < 0.0001). Fet-A levels were positively correlated with insulin resistance (HOMA-IR) (r = 0.34, p < 0.0001) and negatively correlated with insulin sensitivity (QUIKI) (r = -0.41, p < 0.0001).Cu, Se, Zn, and Fe levels were significantly lower in diabetic patients than in non-diabetic patients (p < 0.05). Se and Zn were significantly correlated with Fet-A (r = -0.41, p = 0.049 and r = -0.42, p = 0.001, respectively). Se and Zn were also correlated with insulin resistance (HOMA-IR) (r = -0.45, p = 0.049 and r = -0.36, p = 0.012, respectively) and insulin sensitivity (QUIKI) (r = 0.49, p = 0.042 and r = 0.30, p = 0.003, respectively). Similarly, Fe was negatively correlated with insulin levels (r = -0.33, p = 0.04) and insulin sensitivity (r = -0.34, p = 0.30). However, Mn was significantly correlated with Fet-A (r = 0.37, p = 0.001) and insulin resistance/sensitivity (r = 0.24, p = 0.026 and r = -0.24, p = 0.041) respectively in the diabetic obese group. Mg was an independent predictor of diabesity. Conclusions: Mg play a significant role in obesity-related T2DM pathogenesis and complications via Fet-A, insulin sensitivity, and resistance modifications.

RNA profile of immuno-magnetically enriched lung cancer associated exosomes isolated from clinical samples.

Exosomal cargo secreted from cancer cells has been associated with the development and progression of the tumour. Enriching clinically relevant tissue-specific exosomes may assist in the focused analysis of RNA molecules packaged during cancer. Therefore, this study utilized a rapid immunomagnetic enrichment approach for targeted isolation of lung cancer cell-derived exosomes from human plasma, followed by analysing their cargo RNA using high throughput sequencing. The total RNA purified from these immunomagnetically enriched exosomes provided adequate RNA quality for characterization through the Illumina platform. Differential expression analysis was performed between patients and healthy controls to study the altered exosomal RNA profile during lung cancer. Further, functional enrichment analysis was performed with the list of identified differentially expressed genes (DEGs). In total, 1383 mRNAs and 64 lncRNA were identified as differentially expressed between patient plasma exosomes than healthy controls (fold change > 2, P < 0.05). Kyoto encyclopaedia of Genes and Genomes (KEGG) pathway analysis revealed that the DEGs were mainly associated with cancer-related pathways, purine metabolism, calcium, and cGMP-PKG signalling pathways. In conclusion, the presented approach successfully demonstrated a novel strategy for focused disease-specific transcriptome analysis, which provides a feasible option for identifying disease-specific exosome biomarkers for detecting non-small lung cancer.

Rapid immunomagnetic co-capture assay for quantification of lung cancer associated exosomes.

Exosomes derived from biological fluids have the potential to serve as a biomarker for the early detection of various cancers. However, the lack of reliable enrichment and detection methods posed a challenge for its clinical utility. In this work, we designed a rapid co-capture-based approach for targeted enrichment and detection of lung cancer-derived exosomes from human plasma. This method relies on the formation of a sandwich complex around the exosomes that involves magnetic nanoparticles coupled to CD151 to assist in the immunomagnetic selection of lung-derived exosomes and a secondary detection antibody (CD81) coupled to horseradish peroxidase for signal amplification. The performance of the co-capture method to detect exosomes has been optimized with known exosome concentrations in human plasma and exhibited good linearity (108-105 exosomes mL-1) with a detection limit of 60.4 exosomes µL-1. This study further investigated the potential of the developed assay to differentiate healthy and lung cancer patients using 18 clinical samples by quantifying the CD151+/ CD81+ lung-derived exosomes. In conclusion, this study demonstrated a rapid co-capture-based approach that offers simultaneous isolation and detection of exosomes compatible with low sample volume for detecting lung cancer patients.

Maternal footprints of Southeast Asians in North India.

We have analyzed 7,137 samples from 125 different caste, tribal and religious groups of India and 99 samples from three populations of Nepal for the length variation in the COII/tRNA(Lys) region of mtDNA. Samples showing length variation were subjected to detailed phylogenetic analysis based on HVS-I and informative coding region sequence variation. The overall frequencies of the 9-bp deletion and insertion variants in South Asia were 1.9 and 0.6%, respectively. We have also defined a novel deep-rooting haplogroup M43 and identified the rare haplogroup H14 in Indian populations carrying the 9-bp deletion by complete mtDNA sequencing. Moreover, we redefined haplogroup M6 and dissected it into two well-defined subclades. The presence of haplogroups F1 and B5a in Uttar Pradesh suggests minor maternal contribution from Southeast Asia to Northern India. The occurrence of haplogroup F1 in the Nepalese sample implies that Nepal might have served as a bridge for the flow of eastern lineages to India. The presence of R6 in the Nepalese, on the other hand, suggests that the gene flow between India and Nepal has been reciprocal.

Colorimetric sensing of malathion using palladium-gold bimetallic nanozyme.

In this work, a simple, sensitive and selective label free colorimetric assay using palladium-gold nanorod as nanozyme is reported for malathion detection. Study investigates the peroxidase potential of the nanozyme on colorimetric substrates and explores the effect of selected organophosphates on their enzyme mimetic activity. Palladium-gold nanozyme shows excellent peroxidase mimetic activity with O-phenylenediamine in the presence of hydrogen peroxide. Its Kinetic parameters Km and kcat are better than horseradish peroxidase which makes it a superior enzyme. Nanozyme is stable over a broad temperature range (4-70°C) and shows high peroxidase activity from 2 to 6pH. The peroxidase activity of nanozyme is selectively quenched with increasing concentration of malathion and is the principle of developed assay. Assay has a lowest detection limit of 60ng/ml and shows no cross-reaction with other analogous organophosphates or metal salts. Validation on tap water samples spiked with different concentrations of malathion shows good recovery in the range of 80-106%. Assay also displays good intra and inter-assay precision which lie in the range of 2.7-6.1% and 3.2-5.9% respectively. This study demonstrated the catalytic potential of palladium-gold nanorods, which can be employed as nanozyme for developing highly sensitive detection methods.

Study of a novel method to assist in early reporting of sepsis from the microbiology laboratory.

INTRODUCTION: Microbiology laboratories must provide accurate blood culture reports with rapid turnaround time (TAT) to effectively manage patients with sepsis. In this study three methods are compared for reporting blood culture results: a manual method that included use of a serum separator tube (SST), the conventional manual, and an automated method for identification and susceptibility (ID/AST). METHODOLOGY: Broth from positive blood culture bottles was added to an SST and then centrifuged. The pellet obtained was used to directly inoculate biochemical tests for identification and agar plates for AST on the first day of positivity. Biochemicals and AST plates were read the next day and final results reported on the second day at 24 hours. For conventional disk diffusion testing, the newly positive blood culture broth was also inoculated on solid media on the first day and incubated overnight. The next day AST by was performed as well as biochemical tests from pure colonies. These colonies were also used to inoculate panels for ID/AST using the automated MicroScan 40SI System. These results were recorded on the third day and results reported at 48 hours. RESULTS: The study included 851 samples Out of 106 (12.4%) positive blood cultures, 102 were included in the study; Comparison of the 3 methods showed good correlation. Identification was correctly reported in 95 (93.1%) isolates. The overall AST error rate was 3.8%. CONCLUSIONS: The use of SST and direct from pellet inoculation reduced TAT for identification and AST results between 18 and 24 hours.

Electron paramagnetic resonance spectroscopy in radiation research: Current status and perspectives.

Exposure to radiation leads to a number of health-related malfunctions. Ionizing radiation is more harmful than non-ionizing radiation, as it causes both direct and indirect effects. Irradiation with ionizing radiation results in free radical-induced oxidative stress. Free radical-mediated oxidative stress has been implicated in a plethora of diseased states, including cancer, arthritis, aging, Parkinson's disease, and so on. Electron Paramagnetic Resonance (EPR) spectroscopy has various applications to measure free radicals, in radiation research. Free radicals disintegrate immediately in aqueous environment. Free radicals can be detected indirectly by the EPR spin trapping technique in which these forms stabilize the radical adduct and produce characteristic EPR spectra for specific radicals. Ionizing radiation-induced free radicals in calcified tissues, for example, teeth, bone, and fingernail, can be detected directly by EPR spectroscopy, due to their extended stability. Various applications of EPR in radiation research studies are discussed in this review.

Mitigation approaches to combat the flu pandemic.

Management of flu pandemic is a perpetual challenge for the medical fraternity since time immemorial. Animal to human transmission has been observed thrice in the last century within an average range of 11-39 years of antigenic recycling. The recent outbreak of influenza A (H1N1, also termed as swine flu), first reported in Mexico on April 26, 2009, occurred in the forty first year since last reported flu pandemic (July 1968). Within less than 50 days, it has assumed pandemic proportions (phase VI) affecting over 76 countries with 163 deaths/35,928 cases (as on 15(th) June 2009). It indicated the re-emergence of genetically reassorted virus having strains endemic to humans, swine and avian (H5N1). The World Health Organisation (WHO) member states have already pulled up their socks and geared up to combat such criticalities. Earlier outbreaks of avian flu (H5N1) in different countries led WHO to develop pandemic preparedness strategies with national/regional plans on pandemic preparedness. Numerous factors related to climatic conditions, socio-economic strata, governance and sharing of information/logistics at all levels have been considered critical indicators in monitoring the dynamics of escalation towards a pandemic situation.The National Disaster Management Authority (NDMA), Government of India, with the active cooperation of UN agencies and other stakeholders/experts has formulated a concept paper on role of nonhealth service providers during pandemics in April 2008 and released national guidelines - management of biological disasters in July 2008. These guidelines enumerate that the success of medical management endeavors like pharmaceutical (anti-viral Oseltamivir and Zanamivir therapies), nonpharmaceutical interventions and vaccination development etc., largely depends on level of resistance offered by mutagenic viral strain and rationale use of pharmaco therapeutic interventions. This article describes the mitigation approach to combat flu pandemic with its effective implementation at national, state and local levels.

SCAR markers for correct identification of Phyllanthus amarus, P. fraternus, P. debilis and P. urinaria used in scientific investigations and dry leaf bulk herb trade.

The trade in Phyllanthus material as bulk herb is rampant and mainly involves herbaceous species such as Phyllanthus amarus, P. fraternus, P . debilis and P. urinaria. These species are very important in herbal medicines and have varied activities. In India these species grow sympatrically and there are chances of deliberate or ignorant adulteration of crude drugs, lowering the efficiency of the medication for its intended purpose. Secondly, incorrect identification may also lead to erroneous reports on activities/molecules. To overcome this problem in crude drug (dry leaf powder) and compliment morphological identification in live plant, we have developed SCAR markers for all four species. In each species, we selected one fragment as being monomorphic between accessions but differing in size between species. These species-specific fragments were selected, cloned and sequenced. Based on the sequences, primer pairs were designed and amplification conditions standardized. SCAR markers were isolated from population DNA amplification profiles and validated by sequencing. The species-specific SCAR primers could retrieve the same size and sequence of fragments as in the RAPD profile. These fragments are 1150 bp, 317 bp, 980 bp and 550 bp in size for P. amarus, P. fraternus, P. debilis and P. urinaria, respectively. Additional fragments in P. debilis and P. urinaria indicate different alleles. The retrieval of same size and sequence of species-specific unique SCAR markers from the respective accessions (mixed DNA sample of same accessions) indicates the usefulness to study natural hybridization between the species in addition to adulteration.