Objective assessment of cleft lip and palate speech intelligibility using articulation and hypernasality measures.

Assessment of intelligibility is required to characterize the overall speech production capability and to measure the speech outcome of different interventions for individuals with cleft lip and palate (CLP). Researchers have found that articulation error and hypernasality have a significant effect on the degradation of CLP speech intelligibility. Motivated by this finding, the present work proposes an objective measure of sentence-level intelligibility by combining the information of articulation deficits and hypernasality. These two speech disorders represent different aspects of CLP speech. Hence, it is expected that the composite measure based on them may utilize complementary clinical information. The objective scores of articulation and hypernasality are used as features to train a regression model, and the output of the model is considered as the predicted intelligibility score. The Spearman's correlation coefficient based analysis shows a significant correlation between the predicted and perceptual intelligibility scores (ρ = 0.77, p < 0.001).

Lupeol derivative mitigates Echis carinatus venom-induced tissue destruction by neutralizing venom toxins and protecting collagen and angiogenic receptors on inflammatory cells.

BACKGROUND: Echis carinatus bite is a serious threat in South-Asian countries including India, as it causes highest number of deaths and terrifying long-term tissue destruction at the bitten site. Although venom metalloproteinases and hyaluronidases are the suggested key players, studies on the effect of venom on polymorphonuclear cells, peripheral blood mononuclear cells and platelets, and their role in long-term tissue destruction are still in infancy. While, the effect of venom on collagen receptors, integrin α2ß1/GP VI/DDR1 and CX3CR1 chemokine receptor present on these cells is an untouched area. METHODS: Lupeol, lupeol acetate, its synthetic derivatives 2-8 were screened for inhibition of E. carinatus venom induced-hemorrhage in mouse model where compound 8 was found to be the most potent. Further, compound 8 efficiently neutralized venom induced hemorrhage, edema, dermonecrosis, myonecrosis, myotoxicity, pro-coagulant, oxidative stress, inflammatory cytokines and cleavage of collagen and CX3CR1 receptors on inflammatory cells in in vivo, in silico, ex vivo and in vitro studies. CONCLUSIONS: This study for the first time demonstrated the cleavage of collagen receptors and the receptor for angiogenesis and wound healing by the venom and its inhibition by compound 8, as these are important for firm adhesion of inflammatory cells at the damaged site to resolve inflammation and promote tissue repair. GENERAL SIGNIFICANCE: This study provides a lead in venom pharmacology, wherein, compound 8 could be a therapeutic agent for the better management of viper venom-induced long-term tissue destruction.

Comparison of Bond Strength of Brackets with Foil Mesh and Laser Structure Base using Light Cure Composite Resin: An in vitro Study.

BACKGROUND AND OBJECTIVES: The purpose of this in vitro study was to evaluate the bond strength of the laser-etched base bracket, site of bond failure, and evaluate for enamel remnants on the bracket base after debonding, when compared to foil mesh base bracket. MATERIALS AND METHODS: Sixty noncarious, human premolar extracted for the orthodontic treatment were used for this study. The teeth were randomly divided into two groups containing 30 teeth each, which were bonded with laser-etched base bracket and mesh base bracket using light cure resin. The tensile and mechanical bond strength was tested after 24 hours using TIRA. The forces recorded during debonding were measured in Newton and final readings were tabulated in megapascals (MPa). After debonding, the amount of residual adhesive and enamel detachment on the bracket base were assessed according to adhesive remnant index (ARI) and enamel detachment index (EDI) using stereomicroscope and energy dispersive X-ray spectrometer. RESULTS: The laser-etched base bracket showed statistically significant higher results than mesh base bracket. Mann-Whitney test indicated that laser-etched base bracket had significantly higher tensile bond strength of 8.47 MPa (SD ± 0.84), fatigue strength of 7.75 MPa (SD ± 0.79) compared to mesh base bracket with tensile bond strength of 5.53 Mpa (SD ± 0.89) and fatigue strength of 5.17 MPa (SD ± 1.15). Adhesive remnant index score indicated that laser-etched base bracket had ARI score of 3 for most of the bracket, when compared to mesh base bracket. This was statistically significant. Enamel detachment index scores indicated that less than 10% of enamel detachment occurred in both the types of brackets, which was not statistically significant. CONCLUSION: Laser-etched base bracket showed superior bond strength, when compared to the foil mesh base bracket. The site of bond failure of these laser-etched base bracket was at the interface of enamel-adhesive and did not induce any significant enamel detachment. Thus, we can conclude that laser-etched base bracket is a promising step toward achieving an ideal bracket base design for successful bonding.

Effect of Self-etch Primer-adhesive and Conventional Adhesive Systems on the Shear Bond Strength and Bond Failure of Orthodontic Brackets: A Comparative Study.

BACKGROUND: Prompt-L-Pop is a sixth generation bonding system contains methacrylated phosphoric acid esters that combine an acidic component for etching the enamel and a primer, is an all-in-one adhesive. This study was undertaken to compare the bonding strength of brackets to enamel with traditional bonding technique and the new Prompt-L-Pop system using the same composite resin. MATERIALS AND METHODS: In this in vitro experimental study, 60 human premolar teeth extracted for orthodontic treatment were collected. The samples were randomly divided into three groups comprising of 20 teeth in each group. Shear bond strength and ARI scores for the specimens were measured. Comparison was done using one way ANOVA and Chi-square test. RESULTS: Fourth generation bonding adhesive system depicted similar bond strength to fifth generation bonding adhesive system. Both fourth and fifth generation exhibited higher shear bond strength as compared to sixth generation bonding adhesive system. CONCLUSION: Fourth and fifth generation exhibited higher shear bond strength as compared to sixth generation bonding adhesive system but the sixth generation has clinically acceptable shear bond strength. Also, it was found that sixth generation leaves less residual adhesive on the tooth after bracket removal.

Melatonin alleviates Echis carinatus venom-induced toxicities by modulating inflammatory mediators and oxidative stress.

Viper bites cause high morbidity and mortality worldwide and regarded as a neglected tropical disease affecting a large healthy population. Classical antivenom therapy has appreciably reduced the snakebite mortality rate; it apparently fails to tackle viper venom-induced local manifestations that persist even after the administration of antivenom. Recently, viper venom-induced oxidative stress and vital organ damage is deemed as yet another reason for concern; these are considered as postmedicated complications of viper bite. Thus, treating viper bite has become a challenge demanding new treatment strategies, auxiliary to antivenin therapy. In the last decade, several studies have reported the use of plant products and clinically approved drugs to neutralize venom-induced pharmacology. However, very few attempts were undertaken to study oxidative stress and vital organ damage. Based on this background, the present study evaluated the protective efficacy of melatonin in Echis carinatus (EC) venom-induced tissue necrosis, oxidative stress, and organ toxicity. The results demonstrated that melatonin efficiently alleviated EC venom-induced hemorrhage and myonecrosis. It also mitigated the altered levels of inflammatory mediators and oxidative stress markers of blood components in liver and kidney homogenates, and documented renal and hepatoprotective action of melatonin. The histopathology of skin, muscle, liver, and kidney tissues further substantiated the overall protection offered by melatonin against viper bite toxicities. Besides the inability of antivenoms to block local effects and the fact that melatonin is already a widely used drug promulgating a multitude of therapeutic functionalities, its use in viper bite management is of high interest and should be seriously considered.

Therapeutic drug-induced platelet apoptosis: an overlooked issue in pharmacotoxicology.

The surfacing of the applied fields of biology such as, biotechnology, pharmacology and drug discovery was a boon to the modern man. However, it had its share of disadvantages too. The indiscriminate use of antibiotics and other biological drugs resulted in numerous adverse reactions including thrombocytopenia. One of the reasons for drug-induced thrombocytopenia could be attributed to an enhanced rate of platelet apoptosis, which is a less investigated aspect. The present essay sheds light on the adverse (pro-apoptotic) effects of some of the commonly used drugs and antibiotics on platelets viz. cisplatin, aspirin, vancomycin and balhimycin. Furthermore, the undesirable reactions resulting from chemotherapy could be attributed at least to some extent to the systemic stress induced by microparticles, which in turn are the byproducts of platelet apoptosis. Thereby, the essay aims to highlight the challenges in the emerging trend of cross-disciplinary implications, i.e., drug-induced platelet apoptosis, which is a nascent field. Thus, the different mechanisms through which drugs induce platelet apoptosis are discussed, which also opens up a new perspective through which the adverse effects of commonly used drugs could be dealt. The drug-associated platelet toxicity is of grave concern and demands immediate attention. Besides, it would also be appealing to examine the platelet pro-apoptotic effects of other commonly used therapeutic drugs.

Crocin, a dietary additive protects platelets from oxidative stress-induced apoptosis and inhibits platelet aggregation.

Platelets are the key players in the development of cardiovascular diseases as the microparticles generated by apoptotic platelets and platelet aggregation contribute actively towards the disease propagation. Thus, the aim of this study was to demonstrate the effect of a phytochemical which can prevent these two processes and thereby project it as a cardio-protective compound. Crocin, a natural carotenoid exhibits a wide spectrum of therapeutic potentials through its antioxidant property. The study demonstrated its effects on cytoplasmic apoptotic events of mitochondrial pathway in platelets. Collagen/calcium ionophore-A23187 stimulated platelets were treated with crocin and endogenous generation of reactive oxygen species (ROS) and hydrogen peroxide (H(2)O(2)) were measured. H(2)O(2)-induced changes in crocin-pretreated platelets such as intracellular calcium, mitochondrial membrane potential (ΔΨm), caspase activity, phosphatidylserine exposure and cytochrome c translocation were determined. Crocin dose-dependently ameliorated collagen- and A23187-induced endogenous generation of ROS and H(2)O(2). It also abolished the H(2)O(2)-induced events of intrinsic pathway of apoptosis. Further, it hindered collagen-induced platelet aggregation and adhesion. The current piece of work clearly suggests its anti-apoptotic effect as well as inhibitory effects on platelet aggregation. Thus, crocin can be deemed as a prospective candidate in the treatment regime of platelet-associated diseases.

Viper venom-induced oxidative stress and activation of inflammatory cytokines: a therapeutic approach for overlooked issues of snakebite management.

BACKGROUND AND OBJECTIVE: The snakebite mortality rate has been significantly reduced due to effective anti-venin therapy. The intravenously infused anti-venom will neutralize free and target-bound toxins but fails to neutralize venom-induced inflammation and oxidative stress, as the antigen-antibody complex itself is pro-inflammatory. Therefore, an auxiliary therapy is necessary to treat secondary/overlooked envenomation complications. MATERIALS AND METHODS: Blood samples from healthy donors were treated with viper venom (100 µg/ml) for 2 h. The venom-induced inflammation, oxidative damage and effect of crocin pre-treatment were determined by assessing the serum levels of cytoplasmic, lysosomal and oxidative stress markers along with pro-inflammatory mediators such as tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6 and cyclo-oxygenase (COX)-2. RESULTS: Significantly increased stress markers, cytoplasmic, lysosomal and extracellular matrix-degrading enzymes as well as the pro-inflammatory mediators TNF-α, IL-1ß, IL-6 and COX-2 indicated increased cellular damage but significantly reduced oxidative damage and inflammation in crocin pre-treated groups. CONCLUSION: The data clearly suggest that venom-induced oxidative stress and inflammation is also responsible for oxidative burst and cell death in the circulation, which may worsen even after anti-venin therapy. Hence, the current study demands a supportive therapy in addition to anti-venin therapy to neutralize the overlooked issues of snakebite.

Alleviation of viper venom induced platelet apoptosis by crocin (Crocus sativus): implications for thrombocytopenia in viper bites.

Viper envenomations are characterized by prominent local and systemic manifestations including hematological alterations. Snake venom metalloproteinases (SVMPs) and phospholipase A2 (PLA2) plays crucial role in the pathophysiology of hemorrhage by targeting/altering the platelets function which may result in thrombocytopenia. Platelets undergo the classic events of mitochondria-mediated apoptotic pathway due to augmented endogenous reactive oxygen species (ROS) levels. The observed anticoagulant effects during viper envenomations could be due to exacerbated platelet apoptosis and thrombocytopenia. Moreover, antivenin treatments are ineffective against the venom-induced oxidative stress; therefore, it necessitates an auxiliary therapy involving antioxidants which can effectively scavenge the endothelium-generated/endogenous ROS and protect the platelets. The present study explored the effects of viper venom on platelet apoptosis and its amelioration by a phytochemical crocin. The study evaluated the Vipera russelli venom-induced apoptotic events including endogenous ROS generation, intracellular Ca(2+) mobilization, mitochondrial membrane depolarization, cyt-c translocation, caspase activation and phosphatidylserine externalization which were effectively mitigated when the venom was pre-treated with crocin. The study highlights one of the less studied features of venom-induced secondary complications i.e. platelet apoptosis and sheds light on the underlying basis for venom-induced thrombocytopenia, systemic hemorrhage and in vivo anticoagulant effect.

Vipera russelli venom-induced oxidative stress and hematological alterations: amelioration by crocin a dietary colorant.

Snakebite is a serious medical and socio-economic problem affecting the healthy individuals and agricultural and farming populations worldwide. In India, Vipera russelli snakebite is common, ensuing high morbidity and mortality. The venom components persuade multifactorial stress phenomenon and alter the physiological setting by causing disruption of the blood cells and vital organs. The present study demonstrates the anti-ophidian property of Crocin (Crocus sativus), a potent antioxidant against viper venom-induced oxidative stress. The in vivo oxidative damage induced by venom was clearly evidenced by the increased oxidative stress markers and antioxidant enzymes/molecules along with the proinflammatory cytokines including IL-1ß, TNF-α and IL-6. Furthermore, venom depleted the hemoglobin, hematocrit, mean corpuscular volume and platelet count in experimental animals. Crocin ameliorated the venom-induced oxidative stress, hematological alteration and proinflammatory cytokine levels. At present, administration of antivenom is an effective therapy against systemic toxicity, but it offers no protection against the rapidly spreading oxidative damage and infiltration of pro-inflammatory mediators. These pathologies will continue even after antivenom administration. Hence, a long-term auxiliary therapy is required to treat secondary as well as neglected complications of snakebite.

Hemostatic interference of Indian king cobra (Ophiophagus hannah) Venom. Comparison with three other snake venoms of the subcontinent.

Unlike Naja naja, Bungarus caeruleus, Echis carinatus, and Daboia/Vipera russellii venoms, Ophiophagus hannah venom is medically ignored in the Indian subcontinent. Being the biggest poisonous snake, O. hannah has been presumed to inject several lethal doses of venom in a single bite. Lack of therapeutic antivenom to O. hannah bite in India makes any attempt to save the victim a difficult exercise. This study was initiated to compare O. hannah venom with the above said venoms for possible interference in hemostasis. Ophiophagus hannah venom was found to actively interfere in hemostatic stages such as fibrin clot formation, platelet activation/aggregation, and fibrin clot dissolution. It decreased partial thromboplastin time (aPTT), prothrombin time (PT), and thrombin clotting time (TCT). These activities are similar to that shown by E. carinatus and D. russellii venoms, and thus O. hannah venom was found to exert procoagulant activity through the common pathway of blood coagulation, while N. naja venom increased aPTT and TCT but not PT, and hence it was found to exert anticoagulant activity through the intrinsic pathway. Venoms of O. hannah, E. carinatus, and D. russellii lack plasminogen activation property as they do not hydrolyze azocasein, while they all show plasmin-like activity by degrading the fibrin clot. Although N. naja venom did not degrade azocasein, unlike other venoms, it showed feeble plasmin-like activity on fibrin clot. Venom of E. carinatus induced clotting of human platelet rich plasma (PRP), while the other three venoms interfered in agonist-induced platelet aggregation in PRP. Venom of O. hannah least inhibited the ADP induced platelet aggregation as compared to D. russellii and N. naja venoms. All these three venoms showed complete inhibition of epinephrine-induced aggregation at varied doses. However, O. hannah venom was unique in inhibiting thrombin induced aggregation.

Over-bite and vertical changes following first premolar extraction in high angle cases.

AIMS AND OBJECTIVES: Orthodontists generally agree that nonextraction treatment is associated with downward and backward rotation of the mandible and an increase in the lower anterior face height (LAFH). They also agree that extraction line of treatment is associated with upward and forward rotation of the mandible and decrease in the LAFH. The intent of this cephalometric investigation was to examine the wedge hypothesis, that the vertical dimension collapses after first bicuspid extraction. The present study was undertaken to evaluate the cephalometric overbite and vertical changes following first premolar extraction in high angle cases. MATERIALS AND METHODS: Forty-five adult patients having high mandibular plane angle, i.e. Gogn--SN more than or equal to 32° having class I molar and canine relation were included. Pre and post-treatment lateral cephalograms were measured and compared to analyze the cephalometric changes. RESULTS: There was no decrease in the overbite and vertical changes following first premolar extraction in high angle cases. CLINICAL SIGNIFICANCE: The facial complex does increase in size with growth, but mandibular plane while moving inferiorly, remain essentially parallel to its pretreatment position due to residual growth and treatment mechanics. CONCLUSION: The study concluded that, There was no decrease in the vertical facial dimension, overbite and mandibular plane angle. However, it should be interpreted with caution, given the small sample size.

Association of temporomandibular joint dysfunction, condylar position and dental malocclusions in Davangere population.

AIMS: To study the association between dental malocclusions and temporomandibular joint dysfunction.To study the association between dental malocclusions and condylar position.To study the association between temporomandibular (TM) joint dysfunction and condylar position. METHODS: The subjects were divided into four groups for dental malocclusions viz. class I malocclusion with or without TM dysfunction, class II division 1 malocclusion with or without TM dysfunction, class II division 2 malocclusion with or without TM dysfunction and class III malocclusion with or without TM dysfunction. Once the patient fulfilled the criteria, the presence or absence of signs of TM dysfunction were elicited from the patient. RESULTS: It shows the association between TM dysfunction signs and left and right condylar positions. It shows the association between TM dysfunction symptom and left and right condylar positions. It shows the association between dental malocclusions and TM dysfunction signs and symptom. It shows the association between dental malocclusions and left and right condylar positions. CONCLUSION: There was an association between TM dysfunction signs and left and right condylar positions. But, there was no association between TM dysfunction symptoms and left and right condylar positions. There was an association between dental malocclusions and TM dysfunction signs. But there was no association between dental malocclusions and TM dysfunction symptoms. There was an association between dental malocclusions and left condylar position, but there was no association between dental malocclusion and right condylar position. CLINICAL SIGNIFICANCE: This study indicates that malocclusions and factors of condylar position should be seen as merely cofactors in the sense of one piece of the mosaic in the multifactorial problem of TM dysfunction. TM dysfunction factors that showed significant effects to various malocclusions through this study . This study shows clinical significance of association of various types of dental malocclusions to different conylar positions and TM dysfunction signs and symptoms. Before treating orthodontic patients, one should evaluate and treat the TM disorders for better prognosis.

Comparison of reproducibility of natural head position using two methods.

UNLABELLED: Lateral cephalometric radiographs have become virtually indispensable to orthodontists in the treatment of patients. They are important in orthodontic growth analysis, diagnosis, treatment planning, monitoring of therapy and evaluation of final treatment outcome. AIM: The purpose of this study was to evaluate and compare the maximum reproducibility with minimum variation of natural head position using two methods, i.e. the mirror method and the fluid level device method. MATERIALS AND METHODS: The study included two sets of 40 lateral cephalograms taken using two methods of obtaining natural head position: (1) The mirror method and (2) fluid level device method, with a time interval of 2 months. Inclusion criteria • Subjects were randomly selected aged between 18 to 26 years Exclusion criteria • History of orthodontic treatment • Any history of respiratory tract problem or chronic mouth breathing • Any congenital deformity • History of traumatically-induced deformity • History of myofacial pain syndrome • Any previous history of head and neck surgery. RESULTS: The result showed that both the methods for obtaining natural head position-the mirror method and fluid level device method were comparable, but maximum reproducibility was more with the fluid level device as shown by the Dahlberg's coefficient and Bland-Altman plot. The minimum variance was seen with the fluid level device method as shown by Precision and Pearson correlation. CONCLUSION: The mirror method and the fluid level device method used for obtaining natural head position were comparable without any significance, and the fluid level device method was more reproducible and showed less variance when compared to mirror method for obtaining natural head position. CLINICAL SIGNIFICANCE: Fluid level device method was more reproducible and shows less variance when compared to mirror method for obtaining natural head position.

The metalloprotease, NN-PF3 from Naja naja venom inhibits platelet aggregation primarily by affecting α2ß1 integrin.

NN-PF3 is a non-toxic, anticoagulant, high-molecular-mass (67.81 kDa) metalloprotease from Indian cobra (Naja naja) venom. In the present study, NN-PF3 was investigated for the mechanism of inhibition of collagen-induced aggregation of human platelets. The complete inhibition of collagen-induced aggregation and partial inhibition of ADP- and epinephrine-induced aggregation has the respective IC(50) of 75 ± 5, 185 ± 10, and 232 ± 12 nM, whereas no inhibition of thrombin-, arachidonic acid-, and ristocetin-induced aggregation of platelets was observed in platelet-rich plasma. Further, native NN-PF3 and EDTA-inactivated NN-PF3 inhibited collagen-induced aggregation of washed platelets with respective IC(50) of 75 ± 4 and 180 ± 6 nM. The higher inhibitory effect of native NN-PF3 compared with EDTA-inactivated NN-PF3 suggests the enzymatic and non-enzymatic mechanism of inhibition. NN-PF3 pretreatment affected the collagen binding but not the fibrinogen, and fibronectin binding of washed platelets in adhesion assay suggested that the collagen receptors are affected. Western blot study using anti-integrin α2ß1 mAb 6F1 suggested that NN-PF3 binds to integrin α2ß1 in a primary structure-dependent manner only and is not cleaved. There was a drastic reduction in the intensity of several intracellular signaling phosphotyrosine protein bands when monoclonal anti-phosphotyrosine antibody was used, suggesting that the major activation pathway of platelets get affected, which occurs through glycoprotein VI. NN-PF3 did not bind to collagen as revealed by Western blot using anti-collagen mAb. Furthermore, neither the proteolytic cleavage of fibrinogen nor its degradation products by NN-PF3 contributed for the collagen-induced platelet aggregation inhibition.

Anti-venom potential of aqueous extract of stem bark of Mangifera indica L. against Daboia russellii (Russell's viper) venom.

Several plant extracts rich in pharmacologically active compounds have shown to antagonize venom of several species. Mangifera indica has been used against snakebite by the traditional healers. However, there is paucity of scientific data in support. In this study, we evaluated the antivenom potential of aqueous extract of stem bark of M. indica against D. russellii venom-induced pharmacological effects such as life myotoxicity, edema, LD50 etc. The extract inhibited the phospholipase, protease, hyaluronidase, 5'nucleotidase, ATPase and alkaline phosphomonoesterase activities with varying IC50 values. It significantly inhibited both metalloproteases and serine proteases activities. Further, the extract significantly reduced the myotoxicity of the venom, as evident by the reduction of serum creatin kinase and lactate dehydrogenase activities. Though the extract completely inhibited in vitro PLA2 activity, it was unable to completely inhibit in situ hemolytic and in vivo edema-inducing activities, usually brought about by PLA2s. In lethality studies, co-injection of the venom preincubated with the extract showed higher protection than the independent injection of venom, followed by the extract in the mice. However, in both the cases the extract -a cocktail of inhibitors significantly increased the survival time, when compared to that of mice injected (i.p) with the venom alone. These results encourage further studies on the potential use of cocktail of inhibitors in improving the treatment of snake envenomation. Further, this study substantiates the use of M. indica as an antidote against snakebite by the traditional healers.

Factor Xa-like and fibrin(ogen)olytic activities of a serine protease from Hippasa agelenoides spider venom gland extract.

In the recent past, a low molecular mass serine protease, the Hag-protease that caused pro-coagulant activity and as well as local toxicity was isolated and characterized from the Hippasa agelenoides spider venom gland extract (Devaraja et al., Toxicon 52:130-138, 2008). In the current study, the pro-coagulant property has been investigated further and the results are presented. The Hag-protease reduced the re-calcification time of citrated human plasma. It reduced the activated partial thromboplastin time (APTT), and prothrombin time (PT) suggesting its participation in common pathway of blood coagulation. Interestingly, it coagulated the citrated human plasma in the absence of CaCl(2) but, it was lacking thrombin-like activity as it did not clot the purified fibrinogen. Strikingly, the enzyme coagulated the factor X deficient congenital human plasma, suggesting the factor Xa-like activity. However, the cumulative augmented activity was observed in presence of CaCl(2) and phospholipids. Further, the Hag-protease preferentially hydrolyzed the Aalpha chain and then the Bbeta-chain, but not the gamma-chain. As a result, truncated fibrinogen generated was lacking in the polymerization property. It hydrolyzed all the subunits of partially cross-liked fibrin clot (alpha-polymer, alpha-chain, beta-chain, and gamma-gamma dimers). Further, at low concentrations, the Hag-protease stimulated the aggregation of human platelets in platelet rich plasma, but at high concentrations caused spontaneous clumping. In contrast, it inhibited the collagen induced aggregation of washed human platelets. In summary, the present study for the first time reporting the factor Xa-like activity of a serine protease especially from the spider venom that exhibited opposing effects on hemostasis, the pro-coagulant activity and the anti-coagulant activity including fibrin(ogen)olytic and platelet aggregation inhibition activities.

Hyaluronidase inhibitors: a biological and therapeutic perspective.

The hyaluronidases (HAases) are a group of less extensively studied glycosidases distributed throughout the animal kingdom and are popularly known as 'spreading factors'. In recent years, HAases received much attention due to their ability to abruptly alter the hyaluronic acid (HA) homeostasis. HAases preferentially degrade HA, which is a megadalton acidic structural polysaccharide found exclusively in the extracellular matrix (ECM) of animals. The HA-HAase system has been suggested to participate in many pathophysiological conditions. The HA degradation in ECM, crack down the structural integrity with an eventual increased tissue permeability that is attributed for the spreading property. The spreading property has been widely accepted in functions including envenomation, acrosomal reaction/ovum fertilization, cancer progression, microbial pathogenesis such as wound infections, pneumonia, and other sepses like, bacteremia and meningitis. HA fragmentation has dual effects; generation of a wide molecular range bioactive oligosaccharides of angiogenic, pro-inflammatory, and immunostimulatory properties; and impairment in the reservoir capacity of ECM that holds metal ions, growth factors, cytokines and various enzymes for signal transduction. Hence, inhibition of HA degradation appears critical and imperative in HAase mediated pathological conditions. HAase inhibitors are thus potent regulators that maintain HA homeostasis and they might serve as anti-inflammatory, anti-aging, anti-microbial, anticancer and anti-venom/toxin and contraceptive agents. In addition, HAase inhibitors may serve as tools to understand several unexplained and complex functions of HAases in HA metabolism. Therefore, this review is expected to provide an integrated update as of 2008 on the HAase inhibitors and their possible role as therapeutics in the management of a wide range of pathological conditions.

Effect of anticoagulants on the plasma hyaluronidase activities.

Mammalian hyaluronidases (HAases) are an endo-beta-N-acetyl-hexosaminidases that degrade hyaluronan (HA) and have been implicated in diverse pathophysiological functions. Several pathological conditions, such as diabetes, monoclonal gammapathy, and bladder and prostate tumors, report the distorted plasma HAase activity. However, the plasma HAase (hHyal-1) activity has been presumed to change with the circulating HA level and serves as an early marker for several diseases. It has been generally practised to use the anticoagulants such as tri-sodium citrate/di-sodium EDTA/heparin for the preparation of plasma for both biochemical and clinical analyses. In the present investigation, the effect of anticoagulants on plasma HAaseactivity was evaluated and compared with the serum HAase activity that is devoid of anticoagulants as no study provides information in this regard. The results suggested that the plasma HAase activity in the presence of the recommended concentration of EDTA was highly comparable/similar to that of the serum HAase activity. In contrast, citrated or heparinized plasma recorded a significantly reduced level of activity than that of the serum HAase activity. In conclusion, our results suggested that the EDTA-treated plasma samples are a better choice compared with heparin and citrated samples to assess the HAase activity.

Ethyl 6-(4-chloro-phen-yl)-4-(4-methoxy-phen-yl)-2-oxocyclo-hex-3-ene-1-carboxyl-ate.

In the title compound, C(22)H(21)ClO(4), the cyclo-hex-3-ene unit adopts an envelope conformation in both independent mol-ecules comprising the asymmetric unit. The two benzene rings are inclined to each other at a dihedral angle of 82.03 (5)° [86.37 (5)°]. In the crystal, the molecules interact via C-H⋯O, C-H⋯Cl and C-H⋯π interactions.