Comparative assessment of ISSR, RAPD, and SCoT markers for genetic diversity in Clerodendrum species of North East India.

Clerodendrum belonging to the family of Lamiaceae is used in indigenous systems of medicine to treat various life-threatening diseases. The genus has complex morphological variations which lead to limits in its precise taxonomic classifications. Genetic diversity study could enhance taxonomic authentication and evolutionary relationship among the species of Clerodendrum. In this study, nine species of Clerodendrum collected from different regions of North East India were screened using ISSR, RAPD, and SCoT molecular markers. The markers of ISSR, RAPD, and SCoT generated a total of 79, 126, and 145 amplicons with an average of 6.58, 7.86, and 8.53 amplicon per primer. The polymorphism information contents (PIC) for ISSR, RAPD, and SCoT ranged from 0.28 to 0.37, 0.39 to 0.69, and 0.30 to 0.62 with resolving power (Rp) varying from 5.26 to 11.11, 4.04 to 9.67, and 4.54 to 8.65, respectively. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) based clustering methods grouped 94 genotypes into 6 clusters for ISSR and 3 clusters each for RAPD and SCoT markers. Similarly, population structure-based analysis divided 94 genotypes into 6 populations for ISSR and RAPD and 4 populations for SCoT markers. AMOVA analysis revealed that SCoT markers generated maximum genetic variations within and among genotypes, contrary to ISSR and RAPD markers. Results in this study, suggest that the competence of three markers was relatively the same in genotypes fingerprinting, but SCoT was more efficient in the detection of polymorphism for Clerodendrum species. Further, these results could be integrated in the exploration of diverse Clerodendrum species and germplasm utilization.

Virtual Screening and Network Pharmacology-Based Study to Explore the Pharmacological Mechanism of Clerodendrum Species for Anticancer Treatment.

Background: Cancer is a second leading cause of death in the world, killing approximately 3500 per million people each year. Therefore, the drugs with multitarget pharmacology based on biological networks are crucial to investigate the molecular mechanisms of cancer drugs and repurpose the existing drugs to reduce adverse effects. Clerodendrum is a diversified genus with a wide range of economic and pharmacological properties. Limited studies were conducted on the genus's putative anticancer properties and the mechanisms of action based on biological networks remains unknown. This study was aimed to construct the possible compound/target/pathway biological networks for anticancer effect of Clerodendrum sp. using docking weighted network pharmacological approach and to investigate its potential mechanism of action. Methods: A total of 194 natural Clerodendrum sp. Compounds were retrieved from public databases and screened using eight molecular descriptors. The cancer-associated gene targets were retrieved from databases and the function of the target genes with related pathways were examined. Cytoscape v3.7.2 was used to build three major networks: compound-target network, target-target pathway network, and compound-target-pathway network. Results: Our finding indicates that the anticancer activity of Clerodendrum sp. involves 6 compounds, 9 targets, and 63 signaling pathways, resulting in multicompounds, multitargets, and multipathways networks. Additionally, molecular dynamics (MD) simulations were used to estimate the binding affinity of the best hit protein-ligand complexes. Conclusion. This study suggests the potential anticancer activity of Clerodendrum sp. which could further contribute to scavenger novel compounds for the development of new alternative anticancer drugs.

Botulinum toxin for lower lid entropion correction.

This prospective clinical study was designed to evaluate the efficacy of botulinum toxin for temporary treatment of senile and congenital lower lid entropion. Seventeen patients with senile entropion and three children with congenital entropion were treated with botulinum toxin injection into the preseptal orbicularis muscle of lower lid. This resulted in transient relieve of the condition, which lasted for a period of 8-26 weeks. This technique is easy and effective for senile entropion as well as certain cases of congenital entropion.

DNA barcodes for delineating Clerodendrum species of North East India.

The diversified genus of Clerodendrum with its complex evolutionary history leads to taxonomic mystification. Unlike traditional taxonomic methods, DNA barcoding could be a promising tool for the identification and conservation of Clerodendrum species. This study was attempted to develop an efficient barcode locus in Clerodendrum species of North East India. We evaluated four barcode candidates (ITS2, matK, rbcL, ycf1) and its combinations in different Clerodendrum species. The reliability of barcodes to distinguish the species were calculated using genetic pairwise distances, intra- and inter-specific diversity, barcode gap, and phylogenetic tree-based methods. The results exemplify that matK posse's maximum number of variables and parsimony-informative sites (103/100), intra- (0.021 ± 0.001) and inter- (0.086 ± 0.005) specific divergences and species resolution rate (89.1%) followed by ITS2, ycf1, and rbcL. Among the combinatorial locus, ITS2 + matK showed the best species discrimination with distinctive barcode gaps. Therefore, we tentatively suggest that the combination of ITS2 + matK as core barcode for Clerodendrum and converted into quick response (QR) code. Hence, this finding indicates that DNA barcoding could provide consistent resources for species discrimination and resolve taxonomic controversies of the genus as well as set a preliminary assessment toward its biodiversity.

Combined posterior flap and anterior suspended flap dacryocystorhinostomy: A modification of external dacryocystorhinostomy.

BACKGROUND: External dacryocystorhinostomy (DCR) remains a reliable surgical technique for the treatment of obstruction of lacrimal drainage system beyond the common canalicular opening. AIM: To describe a simple modified double flap external DCR technique. MATERIALS AND METHODS: Ninety six consecutive cases of chronic dacryocystitis with or without mucocele were selected irrespective of age and sex. In a modification to routine external DCR, a modified technique was followed, where both anterior and posterior flaps of lacrimal sac and nasal mucosa are created and sutured. Two double armed sutures were used to join the edges of anterior flaps, and elevate them anteriorly to avoid adhesion or apposition with underlying sutured posterior flaps, and to approximate the deep plane of the wound. RESULTS: At the end of average follow-up period of 13 months, we observed 98.9% objective and 96.8% subjective success rates. The average operation time was 45 minutes. No significant intraoperative or postoperative complications were noticed. CONCLUSION: We believe that combined posterior flap and anterior suspended flap DCR technique is simple to perform and has the advantage of both double flap DCR and anterior suspension of anterior flaps. The results of the study showed the efficacy of this simple modification.

Resection and arthrodesis of the knee joint for giant cell tumours of bone.

PURPOSE: To evaluate functional outcomes and complications following resection and arthrodesis of the knee for giant cell tumours (GCTs) of bone, in comparison to treatment by endoprosthetic replacements reported elsewhere. METHODS: 18 men and 14 women aged 18 to 40 (mean, 28) years underwent resection and arthrodesis of the knee for GCTs of bone involving the distal femur (n=17) and proximal tibia (n=15). After wide resection, 2 struts were fashioned from the harvested fibula/ fibulae and inserted into the medullary canal at the resected ends of the tibia and femur. The corresponding ends of the struts were inserted into peg holes made in the unaffected condyles in a divergent fashion. The knee was arthrodesed in 5 to 10 degrees of flexion, with the limb kept 1 cm short. A 95-degree AO condylar bladeplate (10-12 holes) was fixed at the resected ends, with a minimum of 8 cortices purchase. Cancellous bone grafts were placed transversely along the struts and circumferentially over the host-graft junctions. Outcomes and complications were evaluated and compared with those of endoprosthetic arthroplasty reported elsewhere. RESULTS: Patients were followed up for a mean of 8 (range, 3-12) years. The mean size of the tumours was 10x8x6 cm. All patients achieved arthrodesis and full weight bearing without pain within 6 to 10 (mean, 6) months. No shortening, loss of alignment, loosening, implant breakage ensued. One patient had a deep infection and absorption at the host-graft junction. Another had a stress fracture of the fibular strut after plate removal. Two patients had a transient peroneal nerve palsy. One patient had local recurrence and extensive fungation and underwent amputation. The mean functional score was 26 (87% of the full score), compared to 66 to 85% in endoprosthetic arthroplasty reported elsewhere. CONCLUSION: Arthrodesis is a viable alternative to customised arthroplasty and provides a long-lasting and cost-effective reconstruction for average patients in developing countries.

Calcium phosphate ceramics as bone graft substitutes in filling bone tumor defects.

BACKGROUND: Synthetic bio-inert materials are currently used as an alternative to autogenous bone graft. Calcium hydroxyapatite (HA) and Beta tri-calcium phosphate (beta-TCP), which belong to the calcium phosphate ceramics group, are biocompatible and osteo-conductive. The purpose of this study is to analyse the use of HA and beta-TCP in their ceramic forms as a bone graft substitute in filling bone voids after curettage of benign bone tumors. MATERIALS AND METHODS: Twenty-four patients in the age range of 3.5-55 years (mean 14.3 years) having benign bone tumors with bone defects were filled with bone graft substitute following curettage. In 20 patients bone defects were filled with block/granules of HA ceramic and in four with beta-TCP. Fibular strut graft was packed with HA in four patients. The patients were followed up for an average of 18 months (range 12-36 months). RESULTS: The functional status of the patients at follow-up was evaluated and compared with preoperative functional status. Early incorporation of graft substitutes became evident radiologically between 6 and 10 weeks (Stage I). Complete incorporation (Stage III) was observed in an average of nine months (6-18 months). Clinical healing was observed before radiological healing. The average time taken to return to preoperative function was 14 weeks. There was no recurrence of lesion or growth retardation. CONCLUSION: Calcium hydroxyapatite and beta-TCP are excellent bone graft substitutes for autogenous bone graft in filling voids after curettage of benign bone tumors.