HYALURONIDASE has a panoramic use in biotechnology processes and therapy due to its therapeutic, pathophysiological, physiological and biological importance. Since much of the preparations of HYALURONIDASEs are from animal source (bovine and ovine testicular sources) with limited sources of microbial origin, that prompted the authors to screen and isolate a new promising bacterial strain with higher yield followed by its characterization employing detailed taxonomic studies. The newly isolated strain was identified based upon their micro- and macro-morphological, cultural, physiological and biochemical parameters. Twenty isolates from different pathological samples were primarily selected and further screened for their HYALURONIDASE producing capabilities by measuring reduction in turbidity and hydrolyzed zone of substrate hyaluronic acid. Four isolates showing marked reduction in turbidity (A600 nm) and hydrolyzed zones were selected and subjected to secondary screening by shake flask fermentation. Isolate SII9 (Dental caries specimen) exhibited maximum HYALURONIDASE activity (117 U/ml) when compared to the reference Streptococcus mitis MTCC*2695 (106 U/ml). A close scrutiny of the literature revealed that the characteristics of our isolate SII9 are mostly identical to S. equi subsp. equisimilis with few differences and thus designated as S. equi SED 9.

Background and Objective: Snake venoms comprise complex mixtures of enzymatic and non-enzymatic proteins and small organic compounds. HYALURONIDASE is a constant factor in the venom of the snake and is known as a spreading factor. This factor facilitate spread of toxins that harm into the blood circulation system.In this paper, we describe the determination of HYALURONIDASE activity in Iranian Vipera lebetina venom.Subjects and Methods: HYALURONIDASE activity was assayed turbidometrically on hyaluronic acid. Turbidity reducing activity was expressed as a percentage of the hydrolysed hyaluronate, taking the absorbance of a tube as 100% in which no enzyme was added. Bovin testis HYALURONIDASE was used as standard.Results: The optima PH and temperature for HYALURONIDASE maximum activity was 6 and 37oC respectively.Conclusion: In conclusion the Iranian Vipera lebetina venom possessed considerable HYALURONIDASE activity.

Background: Streptococcus pyogenesproduce extracellular HYALURONIDASE enzyme which is directly associated with the spreading of the organism during infection. HYALURONIDASE enzyme is able to break hyaluronic acid or interstitial cement. This enzyme might be used in cancer treatment.The objective of the present study was to clone and express the nucleotide sequence of this enzyme which is involved in HYALURONIDASE enzymatic activity.Materials and Methods: The enzymatic region of HYALURONIDASE gene was detected by bioinformatics methods. The polymerase chain reaction method was used to amplify the region. The amplified product was cloned into the expression vectorpET32a. E. coli BL21 (DE3) pLYsS was transformed with recombinant plasmids. Then gene expression was induced by IPTG. The expressed protein was purified successfully via affinity chromatography by NiNTA kit. The integrity of the product was confirmed by western-blot analysis.Results: The nucleotide sequence of amplified gene was consistent with the streptocuccal HYALURONIDASE gene. The concentration of recombinant protein calculated to 500 mg purified protein per liter. The enzymatic region of recombinant protein from Streptococcus pyogenes was recognized by all five patient’s sera with Streptococcus infection.Conclusion: In general, it is possible to produce the enzymatic regions of the Streptococcus pyogenesHYALURONIDASE in Escherichia coli. The antigenic property of the produced protein is well retained. Considering the product's domestic demand and also low efficiency of production and pathogenicity of Streptococcus species, it is possible to produce it as recombinant product.

Background: The Scorpion Mesobuthus Eupeus belongs to the Buthidae family. It is one of the most abundant scorpions, present in Khuzestan which is responsible for about forty five percent of stinges in this area. The venom of scorpions is composed mainly of a variety of neurotoxins. These neurotoxins specifically interact with ionic channels of excitable cell.Because of the importance of scorpion in this geographic area and since there was not much work done on the enzyme activity on Mesobuthus eupeus eupeus, we decided to do fractionation of Mesobuthus Eupeus eupeus venom with gel filtration chromatography and to study enzymeactivity in all collected fractions.Materials and Methods: In this study the venom of Mesobuthus Epeus epeus was prepared in lyophilized form. One gram of crude venom was dissolved in 10 ml distilled water and centrifuged for 12 minute in 18000xg. The supernatant containingprotein was applied to the sephadex G-50 gel filtration column chromatography, equilibrated with 20m Mammonium acetate pH 4.7 with flow rate of 30 ml/h. Eluant was collected in 3 ml fractions by fraction collector system. Six fractions were separated. Absorbance of eluants was measured continuously in 280 nm. All fractions were tested for protein content, phospholipase A2 and HYALURONIDASE activity.Results: In crude venom there was 816mg protein. The activity of phospholipase A2 with indirect hemolysate in crude venom and all fractions showed that crudec venom, Fraction I and Fraction II had 53.12, 76.30 and 2.43% hemolysis activity respectively. HYALURONIDASE specific activity was 740 and 4574.5TRU/mg (Turbidity Reducing Unit) in crude venom and in fraction I the fold of purification of HYALURONIDASE was 6.18 by gel filtration chromatography.Conclusion: It was found that phospholipase and HYALURONIDASE activities are present in Mesobuthus eupeus eupeus venom and both are separated in fraction I using gel filtration Chromatography.

Background: HYALURONIDASE A is an antigenic protein that is secreted by Streptococcus pyogenes. Nowadays, streptococcal infections are diagnosed by tracking down anti-HYALURONIDASE A antibodies. In this study, the attempt was made to generate recombinant HYALURONIDASE A in E. coli. Materials and Methods: In this experimental study, through designing specific primers and polymerase chain reaction (PCR), HYALURONIDASE A gene was amplified and after purification, it was sub-cloned in plasmid expression vector pET32a. Then pET32a-hylA was transferred to E. coli BL21- DE3-plySs. Protein generation induced by IPTG. The recombinant protein was purified by Ni-NTA kit and its concentration was assayed by Bradford method. Western-Blot analysis was run for verifying the recombinant HYALURONIDASE A.Results: The nucleotide sequencing of the gene amplified by PCR was the same as HYALURONIDASE A gene from Streptococcus pyogenes. Production of the recombinant HYALURONIDASE A via induction by pET32a-hylA plasmid was done through IPTG. The expressed protein was purified by affinity chromatography by Ni-NTA resin. The concentration of purified protein was 500mg/ml. analysis using a mouse anti-HYALURONIDASE A serum was reacted with the generated protein using Western-Blot analysis.Conclusion: Recombinant HylA protein can be generated in E.coli and the resulting protein maintains its antigenic properties desirably.

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system. Among the numerous proposed etiologies, Borrelia burgdorferi (a causative agent of Lyme disease) has been associated with MS. Although the current MS therapies decrease the quantity and severity of the attacks, most patients experience various neurologic symptoms obliging them to have recourse to one or more complementary and alternative medicines along with the conventional medical interventions.Among these, bee venom (BV) therapy is increasingly used for the treatment of MS; nonetheless no animal or human studies have so far revealed an improvement in the symptoms of MS upon such therapy. Herein, the authors discuss the plausible factors giving rise to the inefficacy of BV in amelioration of MS symptoms, despite its highly anti-inflammatory properties.We hypothesize that BV compound purified of phospholipase A2 that highly contains melittin and HYALURONIDASE may alleviate the symptoms of MS, directly through anti-inflammatory effects and degradation of hyaluronan accumulated in inflammatory demyelinating lesions, and indirectly by inhibitory effects on Borrelia burgdorferi.Thus, upon this hypothesis, we suggest that the melittin and HYALURONIDASE be injected into specific trigger points in the patients diagnosed with MS in randomized clinical trials to assess the efficacy of the proposed modality.

Objective: To analyze the effects of HYALURONIDASE and hirudoid treatment on drug extravasation in neonates.Methods: The medical records of 13 neonates with drug extravasation treated with HYALURONIDASE and hirudoid between August 1st, 2010 and May 1st, 2012 were analyzed retrospectively. The treatment procedure for drug extravasation adhered to the protocol in neonatal department. The information including age, sex, weight, diagnosis, size of affected area, site of extravasation and treatment was collected.Findings: The extravasation injuries alleviated and the symptoms improved after treatment, no adverse drug effects were reported with use of HYALURONIDASE and hirudoid.Conclusion: The treatment appeared to be beneficial in the management of extravasations of various medications in neonates and may be useful in reducing the severity of cutaneous toxicosis. However, further studies with large samples are still needed to assess the effectiveness and safety of HYALURONIDASE and hirudoid.

Purpose: To the assess sensitivity and specificity of urinary levels of hyaluronic acid (HA) and HYALURONIDASE (HAase) as an individual or a combined test to diagnose bladder transitional cell carcinoma (TCC). Materials and Methods: One hundred and ninety-four urine specimens were collected from individuals between July 2007 and March 2008. The urinary level of hyaluronic acid (HA) was measured by Enzyme-linked immunosorbent assay. Thereafter, the urinary levels of HA and HAase were normalized to urinary creatinine level and expressed as ng/mg and μ /mg. Results: Eighty percent of patients with bladder cancer had urinary HA level < 500 ng/mg, and 90% of controls showed HA level < 500 ng/mg (P <. 001). The mean urinary levels of HA in controls did not vary significantly (P <. 05), whereas they significantly increased (2. 5 to 6. 5 folds) in all grades of TCC. More than 80% of patients with grades 2 and 3 TCC had urinary HAase level < 10 μ /mg and over 80% of controls showed HAase level < 10 μ /mg (P <. 05). HYALURONIDASE levels increased in patients with grades 2 and 3 bladder TCC. Conclusion: Measurement of urinary levels of HA and HAase (with 89% sensitivity and 83% specificity) appears to be a highly accurate and non-invasive method for detecting bladder TCC and evaluating its grade.

Background and aims: Maintaining low back pain after spinal surgeries is one of the pretty challenging problems in treating patients. Various modalities have been introduced for treatment and one of the most effective ways between them, is steroid and other drugs injection into epidural space.Materials and methods: In a double blind RCT study, 25 patient from pain clinic with low back pain due to failed back surgery syndrome were candidate for epidural transforaminal block and divided into case and control groups randomly. The selected drug regimens were as follow: in case group bupivacaine %0.5, triamcinolone 40 mg, hypertonic saline 5%, HYALURONIDASE 1500 IU, and in control group stilled water instead of HYALURONIDASE was used per each nerve root. Pain score, analgesic consumption, neurological examinations and satisfaction were evaluated in recovery and 1, 2, and 4 weeks later.Results: There was no difference in demographic data between 2 groups. Pain score and total analgesic consumption in case group was meaningfully higher 2 and 4 weeks after blockade (p<0.01). Patient satisfaction was higher in case group.Discussion: Adding HYALURONIDASE to transforaminal injectate should be an effective for managing chronic low back pain in failed back surgery syndrome.

Background: The HYALURONIDASEs are the enzymes hydrolyze b-1, 4 glycosidic linkage of hyaluronan. Hyaluronan is a polymer consisting of a repeating disaccharide unit found in cumulus ovuforus complex, semen liquid and other tissue. Addition to hydrolyzing the hyaluronan, HYALURONIDASE can penetrate through the cumulus cells layer that surrounds the oocyte, thus it terms spreading factor.Moreover, it is used to increase the absorption and dispersion of injected drugs. This enzyme used to denude oocytes from surrounding cumulus cells in IVF and ICSI.Materials and Methods: At first step, total mRNA was extracted from testis tissue and cDNA was synthesized.Ph20 coding sequence deleted GPI anchor was amplified by means of specific primers designed based on ph20 special CDS and also contained additional regions encoding His tag and distinctive sequence recognized by entrokinase enzyme. Then, an amplified fragment was inserted into pTZ57R and treated by appropriate restriction enzyme to sub clone into pBudCE 4.1. In this recombinant expression vector, attB region was added for insertion of this construct into genome by phiC31 integrase produced by another vector termed as pCMV-Int.Results: The constructed expression vector was confirmed successfully as verified by sequence analysis. After transfection, culture media was extracted and tested on Granulosa cells. The cell mass was separated effectively that indicate this protein is active.Conclusion: In this study, we produced an appropriate vehicle encoding recombinant HYALURONIDASE for therapeutic approach and recombinant protein for MS and infertility therapy.