Effect of 'Procumbenase' a serine protease from Tridax procumbens aqueous extract on wound healing: A scar free healing of full thickness wounds.

Wound healing involves several cellular and molecular pathways. Tridax procumbens activates genetic pathways with antibacterial, antioxidant, anticancer, and anti-inflammatory properties, aiding wound healing. This study purified Procumbenase, a serine protease from T. procumbens extract, using gel filtration (Sephadex G-75) and ion exchange (CM-Sephadex C-50) chromatography. Characterization involved analyses of protease activity, RP-HPLC, SDS-PAGE, gelatin zymogram, PAS staining, mass spectrometry, and circular dichroism. Optimal pH and temperature were determined. Protease type was identified using inhibitors. Wound-healing potential was evaluated through tensile strength, wound models, hydroxyproline estimation, and NIH 3T3 cell scratch analysis. In incision wound rat models, Procumbenase increased tensile strength on day 14 more than saline and Povidone­iodine. It increased wound contraction by 89 % after 10 days in excision wound models, attaining full contraction by day 15 and closure by day 21. Scarless wound healing was enhanced by 18 days of epithelialization against 22 and 21 days for saline and povidone­iodine. Procumbenase increased hydroxyproline concentration 2.53-fold (59.93 ± 2.89 mg/g) compared to saline (23.67 ± 1.86 mg/g). In NIH 3 T3 cell scratch assay, Procumbenase increased migration by 60.93 % (50 µg) and 60.57 % (150 µg) after 48 h. Thus, Procumbenase is the primary bioactive molecule in T. procumbens, demonstrates scar-free wound healing properties.

Thrombin- and plasmin-like and platelet-aggregation-inducing activities of Plumeria alba L. latex: Action of cysteine protease.

ETHNOPHARMACOLOGICAL RELEVANCE: In folk medicine, parts of Plumeria alba L. are used for the treatment of many diseases, with its latex being used for curing skin diseases and promoting wound healing. AIM OF THE STUDY: This study aimed to study the role of P. alba L. latex in hemostasis and platelet aggregation. MATERIALS AND METHODS: The latex of P. alba L. was processed to remove waxes and enrich protein content, and the final extract was named Plumeria alba L. natant latex (PaNL). PaNL was analyzed for protease activity against casein. The type of protease in PaNL was identified by using protease inhibitors such as E-64, phenylmethylsulfonyl fluoride, ethylenediaminetetraacetic acid, and pepstatin A. Human fibrinogen, fibrin, and collagen types I and IV were subjected to hydrolysis with different concentrations of PaNL. The thrombin-like activity of PaNL was determined by analyzing its fibrinogen-clotting and procoagulant activities. The role of PaNL in platelet aggregation was also investigated. Its hemorrhagic and edema-inducing activities were evaluated in a mouse model. Phytochemical compounds were identified by gas chromatography-mass spectroscopy. RESULTS: The findings of casein/gelatin zymography confirmed that PaNL possesses protease activity. The results of the protease inhibition study indicated the presence of a cysteine-type protease(s) in PaNL. PaNL hydrolyzed the subunits of fibrinogen, fibrin, and collagen types I and IV. Its fibrin-degradation activity indicated that PaNL possesses plasmin-like activity. PaNL induced clotting of citrated human plasma within 3 min of incubation in the absence of CaCl2, indicating the presence of thrombin-like activity, which was further confirmed by the results of the fibrinogen-clotting assay. PaNL induced platelet aggregation in the absence of agonists. There was no hemolytic activity. Mice injected with PaNL did not show edema/ hemorrhagic activity. CONCLUSION: PaNL possesses procoagulant, fibrino(geno)lytic, thrombin- and plasmin-like activities and induces platelet aggregation, which could explain its usage for wound treatment in folk medicine.

Properties of chitin and chitosan extracted from silkworm pupae and egg shells.

Chitin and chitosan from silkworm pupae and egg shells show distinct properties with excellent antimicrobial properties and cytocompatiblity. Spent silkworm pupae and hatched egg shells are discarded as waste but contain valuable carbohydrates, proteins and lipids. Chitosan has excellent antimicrobial properties and is widely used for food, medical and biotechnological applications. In this paper, we report the properties of chitin and chitosan from silkworm pupae and egg shells in comparison to commercially available chitosan. Defatted and deproteinated pupae and shells were demineralized and later deacetylated to form chitosan. Thermal behavior, physical structure, antimicrobial activity and ability to support the attachment and growth of NIH3T3 cells were studied. Chitin and chitosan from both pupae and shells had similar structure and composition. Crystallinity of the pupae chitosan was 48% compared to 38% for egg shell chitosan. Silkworm chitosan showed considerably higher antibacterial and antifungal activity compared to standard. Cells were viable in the presence of pupae and egg shell chitosan in all the concentrations tested. Based on these observations, it can be inferred that silkworm pupae and shells provide a renewable and sustainable source for chitosan with properties suitable for food and medical applications.

Status of DDT and pyrethroid resistance in Indian Aedes albopictus and absence of knockdown resistance (kdr) mutation.

BACKGROUND & OBJECTIVES: Aedes albopictus is one of the vectors for dengue and chikungunya and emergence of pyrethroid resistance in this species could be of a major concern in controlling the vector. This study reports insecticide susceptibility status of Ae. albopictus to DDT and pyrethroids in some Indian populations and status of presence of knockdown resistance (kdr) mutations. METHODS: Three to four day old adult female Ae. albopictus collected from Delhi, Gurgaon (Haryana), Hardwar (Uttarakhand), Guwahati (Assam) and Kottayam (Kerala) were bio-assayed with DDT (4%), permethrin (0.75%) and deltamethrin (0.05%) impregnated papers using WHO standard susceptibility test kit. Mosquitoes were PCRgenotyped for F1534C kdr-mutation in the voltage-gated sodium channel (VGSC) gene. DDT and pyrethroid resistant individuals were sequenced for partial domain II, III and IV of VGSC targeting residues S989, I1011, V1016, F1534 and D1794 where kdr mutations are reported in Ae. aegypti. RESULTS: Adult bioassays revealed varying degree of resistance against DDT among five populations of Ae. albopictus with corrected mortalities ranging between 61 and 92%. Kerala and Delhi populations showed incipient resistance against permethrin and deltamethrin respectively. All other populations were susceptible for both the synthetic pyrethroids. None of the kdr mutations was detected in any of DDT, deltamethrin and permethrin resistant individuals. INTERPRETATION & CONCLUSION: Ae. albopictus has developed resistance against DDT and there is emergence of incipient resistance against pyrethroids in some populations. So far, there is no evidence of presence of knockdown resistance (kdr) mutation in Ae. albopictus.

Study of the anesthetic efficacy of inferior alveolar nerve block using articaine in irreversible pulpitis.

AIM: The purpose of this study was to determine the anesthetic efficacy of inferior alveolar nerve block (IANB) using 4% articaine and 2% lidocaine supplemented with buccal infiltration. MATERIALS AND METHODS: Forty five patients, diagnosed with irreversible pulpitis of a mandibular posterior tooth were included in the study. The first group of 15 patients received 2% lidocaine with 1:200000 epinephrine, the second group 2% lidocaine with 1: 80,000 epinephrine and the third group of 15 subjects received 4% articaine with 1:100000 epinephrine. During the access cavity preparation those patients who complained of pain received an additional buccal infiltration. The percentage of subjects who got profound anesthesia and failure to achieve anesthesia were calculated and tabulated using a visual analog scale. RESULTS: The results revealed that 87% of subjects who received 4% Articaine with 1:100,000 epinephrine got satisfactory anesthesia with inferior alveolar nerve block alone. Only 2 (13%) subjects received an additional buccal infiltration and none of the patients failed to obtain complete anesthesia with articaine. In comparison only 40% of subjects got complete anesthesia with 2% lidocaine with 1:200000 and 60% with 2% lidocaine with 1:80,000. CONCLUSION: It can be concluded that 4% articaine can be used effectively for obtaining profound anesthesia for endodontic procedures in patients with irreversible pulpitis.

A preliminary survey for Wolbachia and bacteriophage WO infections in Indian mosquitoes (Diptera: Culicidae).

Maternally inherited Wolbachia endosymbiotic bacteria are known to induce various kinds of reproductive alterations in their arthropod hosts. It has been proposed that this bacterium can be used as a tool for gene drive system in mosquitoes and also for the reduction of population size and modulating population age structure in order to reduce disease transmission. In the present study, we carried out a survey to determine the prevalence of Wolbachia and its phage WO infection in Indian mosquitoes and classified Wolbachia infection into groups A and B based on extensive polymerase chain reaction assay using Wolbachia specific wsp and orf7 gene primers. Out of 20 fieldcaught mosquito species, eight species have shown to be infected. Singly infected with Wolbachia A was found in two species and B group found in four species, while double infection with AB group were found in two species. All the screened mosquito species with positive Wolbachia infection were also infected with phage WO. The knowledge of variation in Wolbachia and phage WO infection rates and inferred susceptibility to infection among different mosquito genera has fundamental implications for designing and successful application of Wolbachia based vector-borne disease control strategies.

Reduction en-masse of inguinal hernia with strangulated obstruction.

"Reduction en masse of inguinal hernia" means reduction/migration of a hernial sac into the properitoneal space. We report the CT findings in a case of reduction en masse with strangulated obstruction. CT scan demonstrated a hernial sac with fibrous constriction band at the neck, situated in the properitoneal space superior to the inguinal region, causing closed-loop obstruction. The hernial sac contained thickened bowel loop with wall enhancement and fluid suggestive of incarceration/strangulation. We propose to call this, 'The properitoneal hernial sac sign', defined as "Presence of a hernial sac in the properitoneal space (and not in the inguinal/femoral canal) containing an obstructed/incarcerated bowel loop and causing small bowel obstruction" to identify "reduction en masse of inguinal hernia".

Effect of pH on spectral characteristics of P5C-ninhydrin derivative: Application in the assay of ornithine amino transferase activity from tissue lysate.

Currently available method(s) for assaying pyrroline-5-carboxylate (P5C), an important intermediate metabolite of ornithine, proline and glutamate metabolic pathways, are cumbersome or not sensitive enough for microanalysis. The present study involving the synthesis of P5C followed by purity check, molecular mass (amu =113.1) determination by mass spectrometry and spectral characterization of P5C-ninhydrin derivative (λ max: 510 nm) confirmed the authenticity of the preparation. Studies on the effect of pH on spectral characteristics of P5C ninhydrin derivative demonstrated a significant change with respect to λ max (620 nm) and several ∼ 12 fold increase in molar extinction coefficient (ε: 1.96 × 10(5)) in alkaline conditions (pH:7.0-8.0) as compared to the reported Molar ε of 1.65 × 10(4) at max λ 510 nm in ethanolic solution. The modified method, with the improved sensitivity, is adopted for the assay of ornithine amino transferase activity in WBC's/platelets lysate(s) from human blood.