Study of the Role of Salivary Lactate Dehydrogenase in Habitual Tobacco Chewers, Oral Submucous Fibrosis and Oral Cancer as a Biomarker.

AIM: To evaluate the salivary lactate dehydrogenase (LDH) levels in clinico-pathologically confirmed oral submucous fibrosis (OSMF), oral cancer and clinically diagnosed tobacco pouch keratosis patients. MATERIALS AND METHODS: A prospective, comparative study was carried out in a tertiary healthcare centre located in Loni from October 2013 to January 2014. A total of 120 patients were separated into 4 groups depending upon the clinical diagnosis as follows. Group I: healthy control (with no addictions and diseases). Group II: oral cancer. Group III: oral submucous fibrosis. Group IV: habitual tobacco chewers (tobacco addiction without any disease). Substantiation was done using biopsy. The samples were inspected for salivary LDH levels by the technique in line with the recommendations of the International Federation of Clinical Chemistry with the help of Erba Chem semi auto analyser. RESULTS: The mean salivary LDH levels in the control, oral cancer OSMF and habitual tobacco chewer group were 86.12 ± 7.05 IU/L, 592.09 ± 28.57 IU/L, 350.43 ± 5.90 IU/L and 125.19 ± 13.42 IU/L, respectively. Out of 4 groups, LDH activity was increased in saliva of patients with tobacco pouch keratosis, OSMF, and oral cancer consistently. Notable difference was found in the mean salivary levels of the above groups. Results were subjected to appropriate statistical analysis: one-way ANOVA, Student's unpaired t test for group-wise comparison followed by post hoc Tukey's test. CONCLUSION: We observed congruous higher levels of salivary LDH in oral precancer and cancer, and hence it could be a future marker.

Activating Hair Follicle Stem Cells via R-spondin2 to Stimulate Hair Growth.

Wnt signaling is required for the development of the hair follicle, and for inciting the growth (anagen) phase of the hair cycle. Most strategies to enhance Wnt signaling for hair growth create a state of constitutive Wnt activation, which leads to neoplastic transformation of the epithelial hair matrix. Using Axin2(LacZ/+) and Axin2(Cre/+)R26R(mTmG/+) reporter mice and RNA analyses, we show that Wnt signaling is elevated during anagen, is reduced at the onset of catagen, and can be reamplified in the skin and surrounding hair follicles via intradermal injection of recombinant R-spondin2 protein. Using Lgr5(LacZ/+) reporter mice, we demonstrate that this amplified Wnt environment leads to activation of leucine-rich repeat-containing G-protein coupled receptor 5-positive stem cells in the hair follicle. The onset of catagen is repressed by R-spondin2 injection, and the anagen phase persists. As a consequence, hair shafts grow longer. We conclude that R-spondin2 treatment activates hair follicle stem cells and therefore may have therapeutic potential to promote hair growth.

Drugging a stem cell compartment using Wnt3a protein as a therapeutic.

The therapeutic potential of Wnt proteins has long been recognized but challenges associated with in vivo stability and delivery have hindered their development as drug candidates. By exploiting the hydrophobic nature of the protein we provide evidence that exogenous Wnt3a can be delivered in vivo if it is associated with a lipid vesicle. Recombinant Wnt3a associates with the external surface of the lipid membrane; this association stabilizes the protein and leads to prolonged activation of the Wnt pathway in primary cells. We demonstrate the consequences of Wnt pathway activation in vivo using a bone marrow engraftment assay. These data provide validation for the development of WNT3A as a therapeutic protein.

Augmenting endogenous Wnt signaling improves skin wound healing.

Wnt signaling is required for both the development and homeostasis of the skin, yet its contribution to skin wound repair remains controversial. By employing Axin2(LacZ/+) reporter mice we evaluated the spatial and temporal distribution patterns of Wnt responsive cells, and found that the pattern of Wnt responsiveness varies with the hair cycle, and correlates with wound healing potential. Using Axin2(LacZ/LacZ) mice and an ear wound model, we demonstrate that amplified Wnt signaling leads to improved healing. Utilizing a biochemical approach that mimics the amplified Wnt response of Axin2(LacZ/LacZ) mice, we show that topical application of liposomal Wnt3a to a non-healing wound enhances endogenous Wnt signaling, and results in better skin wound healing. Given the importance of Wnt signaling in the maintenance and repair of skin, liposomal Wnt3a may have widespread application in clinical practice.

Dexmedetomidine in a child with methylphenidate intoxication.

Methylphenidate intoxication, due to accidental ingestion, is a common occurrence in pediatrics. Symptoms of extreme agitation are typically controlled with benzodiazepines or barbiturates. There is, however, a legitimate risk of mechanical ventilation due to respiratory depression with increasing doses of benzodiazepines. The authors describe a case of 7-y-old girl with methylphenidate toxicity where dexmedetomidine was successfully used to manage agitation and cardiovascular stimulation without respiratory compromise.

Persistent hiccups associated with epidural ropivacaine in a newborn.

OBJECTIVE: To report a case of persistent hiccups associated with epidural ropivacaine in a newborn infant. CASE SUMMARY: A term female infant (3.05 kg) received epidural ropivacaine for pain control during and after an operative procedure to correct a tracheoesophageal fistula. Three intermittent doses of ropivacaine were administered during the operative period (total dose 2.29 mg/kg) followed by a continuous epidural (caudal) infusion (0.1% ropivacaine; initial dose 0.23 mg/kg/h plus fentanyl 0.46 µg/kg/h). The infant was extubated in the recovery area and transferred to the intensive care unit. Within hours of transfer, she developed persistent hiccups. The epidural infusion was titrated for pain control, up to 0.32 mg/kg/h (ropivacaine). The hiccup frequency increased to every 10-30 seconds, with the patient appearing hypotonic with lip trembling and intermittent tongue fasciculation. An electroencephalogram did not show any epileptiform activity or focal features consistent with seizure activity. The epidural infusion was reduced to 0.26 mg/kg/h (ropivacaine), with dramatic improvement in hiccups and tone. The infusion was discontinued and complete resolution of hiccups was observed. DISCUSSION: Ropivacaine is commonly used for infiltration anesthesia and peripheral and epidural block anesthesia. Use of the Naranjo probability scale determined that our patient's hiccups were probably caused by ropivacaine. To our knowledge, this is the first report of persistent hiccups associated with epidural ropivacaine. CONCLUSIONS: Clinicians should consider the potential of neurotoxicity, manifested as persistent hiccups, when epidural ropivacaine is administered to young infants.

Spermiogenesis in caecilians Ichthyophis tricolor and Uraeotyphlus cf. narayani (Amphibia: Gymnophiona): analysis by light and transmission electron microscopy.

Spermiogenesis, known as spermateleosis in lower vertebrates, is the transformation of the round spermatid into a highly specialized spermatozoon with a species-specific structure. Spermateleosis and sperm morphology of two species of caecilians, Ichthyophis tricolor and Uraeotyphlus cf. narayani, from the Western Ghats of Kerala, India, were studied using light and transmission electron microscopy. Spermateleosis is described in early, mid-, and late phases. During the early phase, the spermatid nucleus does not elongate, but the acrosome vesicle is Golgi-derived and its material is produced as a homogeneous substance rather than as discrete granules. In development of the acrosome, the centrioles shift in position to the lower half of the cell. The acrosomal vesicles take the full shape of the acrosome with the establishment of the perforatorium in midphase. An endonuclear canal develops and accommodates the perforatorium. The incipient flagellum is laid down when the proximal centriole attaches to the posterior side of the nucleus and the distal centriole connects to the proximal centriole, which forms the basal granule of the acrosome. The axial fiber also appears during midphase. The mitochondria shift in position to the posterior pole of the cell to commence establishment of the midphase. Late phase is characterized by nuclear condensation and elongation. Consequently, the final organization of the sperm is established with the head containing the nucleus and the acrosome. The undulating membrane separates the axoneme and axial fiber. Most of the cytoplasm is lost as residual bodies.

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.

Unusual sites (thorax and umbilical hernial sac) of endometriosis.

Extra-pelvic endometriosis in the thorax or umbilical hernia is rare. We report a case of thoracic endometriosis presenting with isolated chest pain and a case of endometriosis in an umbilical hernial sac mimicking incarceration. The clinical course and management of the patients and a literature review of these two unusual sites of endometriosis are discussed.

Clinical activity of the new triazole drug voriconazole (UK 109, 496) against disseminated hepatosplenic aspergillosis in a patient with relapsed leukemia.

There is presently a limited antifungal armamentarium comprising amphotericin and the azoles, fluconazole and itraconazole. In vitro studies have shown efficacy of the new drug, voriconazole, against a wide range of fungi, including most species of Candida and Aspergillus. We review here a case report of a young boy with acute myeloid leukemia who developed disseminated hepatosplenic aspergillosis. He failed therapy with itraconazole, amphotericin B and liposomal amphotericin. As he also had relapsed leukemia, there was a great urgency to control this infection in order to facilitate the administration of cancer chemotherapy. Voriconazole was given with good response resulting in virtual disappearance of all scan evidence of aspergillosis.

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.

Male reproductive toxicity of vincristine: ultrastructural changes in the epididymal epithelial apical cell.

The toxic effect of vincristine on the apical cells of the rat caput epididymis was investigated. The drug was administered at 20 and 40 microg/kg body weight daily for 15 days. Light microscopy using semithin sections, and transmission electron microscopy, of the caput epididymis were undertaken. The results revealed that the basal region of the apical cell was in contact with the basement membrane and the luminal end took part in endocytosis. The apical cells reflected a dose-dependent response to vincristine (VCR) treatment. In general the changes included protrusion of the apical ends deep into the lumen, with the nucleus of the cell located in such protruded ends, and an increase in the abundance of lysosomal bodies and multivesicular bodies. These changes reflected the physiological response of the apical cell to VCR treatment rather than toxic manifestations.

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.