Conservative Management Of Partial-Thickness Scald Burns In Children Using Cultured Allogenic Keratinocyte Spray: Initial Experience Of 18 Patients Treated In An Outpatient Setting.

The objective of this study was to describe our clinical experience with the use of cultured allogeneic keratinocyte (CAK) using a simplified cell delivery method in the treatment of pediatric partial-thickness scald burns treated as outpatients in a Burn Unit. An actuator fitted onto a 3ml syringe was used for cell spray. Eighteen patients having active mixed partial-thickness burn wound areas of <10% total body surface area (TBSA), treated between 2017 and 2019, were included in the study. The wounds were managed conservatively with a combination of burn dressings using hydrogels and CAK application. The timing of the CAK application was decided by the treating plastic surgeon based on his clinical judgment and the clinical status of the wound. The primary study endpoints were the number of days and dressing changes required for complete wound reepithelialization. All of the eighteen patients' wounds reepithelialized completely with CAK application, with a mean reepithelialization time of 10.33 (±4.95) days after the application of CAK. The median value for the number of CAK applications and total dressing sessions required to achieve complete healing were 3 and 4 times, respectively. Wounds treated with CAK application between 8-21 days after burn injury required fewer cell application sessions and fewer dressing changes than wounds treated within seven days and after 21 days from the burn injury. None of the patients reported any adverse reaction related to CAK use. The present study suggests that non-extensive mixed partial-thickness scald burn in children can be successfully treated conservatively using CAK as an adjunct in addition to standard dressing in the outpatient setting.

Le but de cette étude est de décrire notre expérience clinique avec l'utilisation de kératinocytes allogéniques de culture (KAC), en utilisant une méthode simplifiée d'application cellulaire pour le traitement des brûlures intermédiaires de l'enfant par eau chaude, traitées en ambulatoire dans notre unité de brûlés. Un embout, fixé à une seringue de 3 ml, a été utilisé pour la vaporisation cellulaire. 18 patients présentant des brûlures de profondeur intermédiaire au stade aigu, de surface <10% de la surface corporelle totale (SCT) et pris en charge entre 2017 et 2019, ont été inclus dans cette étude. Les plaies ont reçu un traitement conservateur combinant des pansements à base d'hydrogel et l'application de KAC. Le moment pour appliquer les KAC a été choisi par le chirurgien plasticien référent en fonction de son appréciation clinique et du stade de la plaie. Les critères principaux d'évaluation de l'étude ont été le nombre de jours et le nombre de réfections de pansements nécessaires à une réépithélialisation complète. Les plaies des 18 patients ont été totalement réépithélialisées par l'application de KAC, avec un délai moyen de 10,33 (+/-4,95) jours après la vaporisation cellulaire. La valeur médiane du nombre d'applications de KAC d'une part et de réfections de pansements d'autre part, nécessaires pour obtenir une cicatrisation complète a été respectivement de trois applications de KAC et de quatre pansements. Les plaies traitées par l'application de KAC dans un délai de 8 à 21 jours après la brûlure ont nécessité moins de séances d'application cellulaire et de pansements que les plaies traitées dans les sept premiers jours ou au-delà du 21e jour après la brûlure. Aucun des patients n'a présenté d'effets secondaires rapportés à l'utilisation de KAC. Cette étude suggère que les brûlures peu étendues intermédiaires par ébouillantement chez l'enfant peuvent être menées à cicatrisation par un traitement conservateur utilisant la vaporisation de KAC en complément de pansements standards en traitement ambulatoire.

In Vivo and In Vitro Toxicity Profiles of Hexane Extract of Alpinia malaccensis Rhizome in Rat and Cell Line Models.

The objective of the study was to evaluate the potential toxicity of crude n-hexane extract of Alpinia malaccensis rhizome. The in vivo acute oral toxicity was evaluated by administering a single oral dose of the extract at 0, 300, or 2000 mg/kg body weight to female Wistar rats according to modified OECD Test Guideline 423. For the in vitro cytotoxicity study, A549, HepG2, 3T3, and COS-7 cell lines were exposed to different doses of A. malaccensis extract and cell viability was assessed adopting MTT assay followed by AO/EB staining, Hoechst staining, and comet assay with a view to compare the cellular and molecular mechanisms underlying the toxicity, if any. It was found that administration of 2000 mg/kg bw dose in in vivo oral acute toxicity study did not produce significant toxicity or mortality. No significant (p < 0.05) differences were observed for body weight and hematological and biochemical parameters compared to control after 14 days of treatment. No changes in behavior, body weight, hematological and biochemical parameters, and aspects of histopathology were observed when compared to the control. Thus, the possible oral lethal dose for A. malaccensis extract is above 2000 mg/kg body weight. The in vitro cytotoxicity analysis showed nontoxicity concentrations of the extract to be 2, 1.4, 30, and 1.4 µg/mL for A549, HepG2, 3T3, and COS-7 cells, respectively, where no apoptotic/necrotic cell death and DNA damage were observed. In conclusion, the extract of rhizome of A. malaccensis did not produce apparent cytotoxicity or acute oral toxicity, confirming the scope to use A. malaccensis as a safe food preservative and a natural therapeutic product after further subacute and chronic toxicity studies.

Antiproliferative and apoptosis-induction studies of 5-hydroxy 3',4',7-trimethoxyflavone in human breast cancer cells MCF-7: an in vitro and in silico approach.

The aim of this study was to find the efficacy of 5-hydroxy 3',4',7-trimethoxyflavone (HTMF), a flavonoid compound isolated from the medicinal plant Lippia nodiflora, in inhibiting the proliferation and inducing apoptosis in human breast cancer cell line MCF-7. The anti-proliferative effect of the compound HTMF was confirmed using MTT cytotoxicity assay. Increased apoptotic induction by HTMF was demonstrated by acridine orange/ethidium bromide (AO/EtBr) and Hoechst 33258 staining studies. The phosphatidylserine translocation, an early feature of apoptosis and DNA damage were revealed through AnnexinV-Cy3 staining and comet assay. Moreover, the significant elevation of cellular ROS was observed in the treated cells, as measured by 2,7-diacetyl dichlorofluorescein (DCFH-DA). The mRNA expression studies also supported the effectiveness of HTMF by shifting the Bax:Bcl-2 ratio. The treatment of MCF-7 cells with HTMF encouraged apoptosis through the modulation of apoptotic markers, such as p53, Bcl-2, Bax, and cleaved PARP. In silico molecular docking and dynamics studies with MDM2-p53 protein revealed that HTMF was more potent compound that could inhibit the binding of MDM2 with p53 and, therefore, could trigger apoptosis in cancer cell. Overall, this study brings up scientific evidence for the efficacy of HTMF against MCF-7 breast cancer cells.

Tributyltin-mediated hepatic, renal and testicular tissue damage in male Syrian hamster (Mesocricetus auratus): a study on impact of oxidative stress.

Organotin compounds are a versatile group of organometallic chemicals that are used in a variety of industrial and agricultural applications. Tributyltin (TBT), a common organotin, brings about severe spermatotoxic and organotoxic effects. However, information about the adverse effects of TBT on liver, kidney and testis is scanty. Hence, the present study was undertaken to elucidate the TBT-mediated oxidative stress-induced impairments in these organs. Administration of TBT through oral route at increasing doses of 50, 100 and 150 ppm for 65 days to male Syrian hamsters resulted in drastically decreased activities of antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase and decreased mean levels of non-enzymatic antioxidants (reduced glutathione, vitamin C, and vitamin E) followed by a dramatic increase in the levels of lipid peroxidation in the liver, kidney and testis as compared to the control animals. Significantly high levels of serum urea, creatinine, uric acid and bilirubin were observed in TBT-treated hamsters. Also, TBT treatment induced drastic histopathological changes in the liver, kidney and testis combined with remarkable changes in serum levels of tissue injury marker enzymes Aspartate transaminases, Alkaline phosphatase and Alanine transaminase. These data affirm that exposure to TBT can lead to oxidative stress-induced damage to liver, kidney and testis.

Apolipoprotein E Induction in Syrian Hamster Testis Following Tributyltin Exposure: A Potential Mechanism of Male Infertility.

Tributyltin (TBT) is a common environmental contaminant used as the active ingredient in many products such as a biocides, wood preservatives, disinfecting agents, and antifouling paints. The TBT is a known endocrine disruptor. The aim of the current investigation was to determine the toxicity of TBT in the reproductive tract of adult male Syrian hamsters and to ascertain whether this compound results in untoward effects on apolipoprotein E (ApoE), a lipoprotein central to sex hormone synthesis. The TBT was administered orally to male Syrian hamsters at doses of 50, 100, and 150 ppm/kg for 65 days of treatment. We determined body weight, testis weight, sperm count, sperm morphology, testis histology, ApoE expression, serum lipid profile, testosterone level, follicle-stimulating hormone receptor (FSHR), and steroid hormone receptor expression compared to vehicle-treated controls. High doses of TBT significantly affected each of these parameters in Syrian hamsters. Weight and morphology of the testis were altered as well as sperm production. Real-time reverse-transcriptase polymerase chain reaction analysis revealed that expression of ApoE messenger RNA was upregulated in testes from TBT-treated groups compared with controls while the expression of androgen receptor, FSHR, estrogen receptor α (ESR1), and estrogen receptor ß (ESR2) was decreased. We posit that exposure to TBT hinders intracellular cholesterol transport resulting in abnormal sex steroid biosynthesis and subsequent spermatogenic defects. Importantly, these effects may account for the decreased level of normal sperm observed in hamsters exposed to TBT.

Light and electron microscopic observations of fabrication, release, and fate of biphasic secretion granules produced by epididymal epithelial principal cells of the fan-throated lizard Sitana ponticeriana cuvier.

The epididymis of the fan-throated lizard Sitana ponticeriana was examined with light and transmission electron microscopy to understand the cellular mechanisms of fabrication of secretion granules in epithelial principal cells, granule release into the lumen, and the fate of the dense structured granules after reaching the lumen. Principal cells of the ductus epididymis, except at the cauda, secrete electron-dense biphasic granules copiously, which decrease in abundance from the initial segment to corpus. The principal cell possesses a prominent Golgi apparatus and all versions of endoplasmic reticulum (ER), rough, smooth, and sparsely granulated. The material of the dense portion of the secretion granules, after processing at the Golgi apparatus, appears to accumulate in large ER cisternae in the supranuclear cytoplasm. It undergoes condensation when the cisternae become condensing vacuoles. Mitochondria appear to play a role in dense granule formation. The condensing vacuoles are displaced toward the apical cytoplasm when the material of the less dense portion is added to the condensing vacuoles at the Golgi area. Thus, the less dense and dense portions of the secretion granules are secreted and added to the condensing vacuoles separately. The composite granules are released into the lumen by exocytosis when the less dense portion merges with the luminal content, whereas the dense portion maintains its structured identity. The latter, initially measuring 1-2 microm in diameter, increases in size several times. It is inferred that these granules release their content gradually, resulting in the appearance of vacuoles, and suggesting that the granules have an insoluble matrix in which there is a sparingly soluble material. The substance leaching out of the granules appears to contribute to keeping the sperm quiescent and alive during storage in the male reproductive tract.

Ultrastructural evidence for secretion from the epithelium of ampulla ductus deferentis of the fan-throated lizard Sitana ponticeriana Cuvier.

Among reptiles, an ampulla ductus deferentis has been reported only in Squamata. Fairly detailed studies are available only for two species, the lizard Calotes versicolor (Fam: Agamidae) and the snake Seminatrix pygaea (Fam: Colubridae). The light microscopic study on C. versicolor revealed the ampulla to be a prominent organ, whereas the light and transmission electron microscopic study in S. pygaea revealed it to be discernable only in histological preparations. Further, the epithelium of the ductal portion of vas deferens as well as the ampulla of C. versicolor appears to contribute to the seminal plasma and can also phagocytose dead sperm, whereas in S. pygaea neither of these roles has been established. Thus, we hypothesize that there may be variations in the anatomy, histology, and the role of the vas deferens in general, and the ampulla in particular, of the squamate reptiles. In this study, the ductus deferens of the small fan-throated lizard Sitana ponticeriana (Fam: Agamidae) was subjected to light and transmission electron microscopic analysis. In this lizard the ampulla is more prominent than in C. versicolor. The epithelium of the ductal portion of vas deferens consists of principal cells (with features reflecting roles in endocytosis and phagocytosis of dead sperm), dark cells (which are absent in the epithelium of the ductal portion of vas deferens of snakes), and basal cells. The ampulla of S. ponticeriana is differentiated into storage and glandular portions. The epithelium of the storage portion is like that in the ductal portion of the vas deferens, whereas that of the glandular portion, consisting of dark and light principal cells and foamy cells, is tall and forms into smooth villous folds. All three cell types show evidence for a role in secretion, in all likelihood different from each other, for release into the lumen to contribute to seminal plasma. These cells do not provide evidence of a role in phagocytosis of dead sperm. It appears that within the Squamata, the ductal ampulla differs in structure as well as function. We suggest that the ductal ampulla of agamid lizards is a composite gland of the ampulla ductus deferentis and seminal vesicles of mammals.

Male reproductive toxic effects of carbendazim: hitherto unreported targets in testis.

Carbendazim (MBC), a widely used fungicide, is toxic to male reproductive mechanisms. Various cellular targets in the testis for MBC action are being understood only recently and still more targets have been conceived. The present study was aimed at finding such newer targets. Male rats were administered through oral route a single dose of carbendazim (400 mg/kg) and the testis was studied adopting routine histological technique. It has been observed that pachytene spermatocytes could also be targets for MBC action in the testis. The study also reports selective loss of step 14 spermatids, asynchrony of the stages in the spermatogenic cycle and development of Sertoli cell fibrosis of the seminiferous tubules of carbendazim-treated rats. From the different kinds of responses seen in the seminiferous tubules in the same testis to MBC, particularly in the Sertoli cell, MBC action in the testis appears dependent on the stage in the spermatogenic cycle at first exposure.

Spermatotoxic effect of carbendazim.

Carbendazim, suspended in sunflower oil, was administered to Wistar male rats through an oral intubation at a daily dose of 25 mg/kg body weight for 48 days, and the cauda epididymal sperm were analysed on day 49 for counts, motility and abnormalities. The study indicates that carbendazim affects the cauda epididymal sperm as seen in decreased sperm counts, inhibition of motility and increased incidence of abnormalities.

Curative property of Withania somnifera Dunal root in the context of carbendazim-induced histopathological changes in the liver and kidney of rat.

The liver and kidney of rat underwent severe histopathological lesions when treated with a single bolus dose of carbendazim, a fungicide, particularly affecting the hepatocytes and the renal corpuscles, respectively. The effects appear to be manifestations of the microtubule-disrupting activity of carbendazim. Treatment of carbendazim-treated rats with the powder of tuberous root of Withania somnifera (Ashwagandha) for 48 days resulted in complete cure of these organs. The results indicate that Withania somnifera would be an effective curative for carbendazim-induced histopathological changes in the liver and kidney.

Carbendazim generates symplasts in rat spermatogenic clones.

In order to find non-microtubular targets in the seminiferous epithelium for the fungicide and reproductive toxicant carbendazim, it was administered to 90 days old male Wistar rat in a single bolus dose of 400 mg/kg body weight through an oral intubation. A parallel control group was maintained. Rats were sacrificed 48 days after the treatment and the testes were analysed for histopathological changes adopting routine histological methods, when symplasts were localised. The maximum diameter of five largest symplasts was measured, and the number of nuclei in these symplasts was also determined. As it is known that symplasts of spermatogenic cells are produced due to opening up of the intercellular bridges between cells in a clone consequent upon disruption of actin microfilaments, the present study shows that actin microfilaments would also be targets in the seminiferous epithelium for carbendazim toxicity.