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1.
Indian J Otolaryngol Head Neck Surg ; 76(2): 2057-2061, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38566729

ABSTRACT

Inverted papilloma is a tumor found involving the nasal cavity and paranasal sinuses. They are not cancerous but can be locally aggressive. They are most commonly seen involving the lateral wall of nose and maxillary sinus. When the tumor involves the frontal sinus and sphenoid sinus or the post ethmoid cells it presents a surgical challenge. Inverted papilloma almost always occur unilateraly and affect mainly the maxillary sinus. Inverted papilloma arising from the sphenoid and frontal sinuses are rare. Here we present two cases; one involving a 60 year man, with inverted papilloma originating from the frontal sinus and another case where it originated from the sphenoid sinus anterior wall and posterior ethmoidalcell (Onodi cell) and was resected endoscopically.

2.
Indian J Otolaryngol Head Neck Surg ; 76(1): 1421-1424, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38440472

ABSTRACT

Schwannoma is a benign neurogenic neoplasm which is arising from schwann cells of peripheral nerve sheath. It can occur in anywhere in the body. Schwannoma of sinonasal tract is extremely rare. Here we report a rare case of schwannoma of nasal tip. A 45 year old male presented with a swelling of tip of the nose, causing cosmetic deformity with no other associated symptoms. Surgical excision of the mass done by using external rhinoplasty approach. Histolopathology report of specimen showed a well circumscribed schwannoma with Antoni A and Antoni B areas.

3.
J Oral Maxillofac Pathol ; 25(1): 189-192, 2021.
Article in English | MEDLINE | ID: mdl-34349434

ABSTRACT

BACKGROUND: To emphasize the role of odontometric parameters which may be used as a reliable forensic tool especially in cases with partial remains of the individual. AIM: To assess the reliability of odontometric parameters in stature analysis. MATERIALS AND METHODS: The study was conducted on 100 patients (50 male and 50 Female). Mesiodistal width of anterior teeth, inter-canine width, Inter-premolar width and maxillary arch length were the parameters included. The results were tabulated with a linear regression formula obtained for each parameter. STATISTICAL ANALYSIS: The data collected was statistically analysed using SPSS version 20 and a linear regression formula was obtained thereafter. RESULTS: Mesio-distal widths of individual maxillary canines, total mesiodistal width of maxillary anteriors and arch length showed a significant co-relation with stature. The combined linear regression formula was obtained for all parameters. CONCLUSION: Odontometric parameters of mesio - distal widths of individual maxillary canines, total mesiodistal width of maxillary anteriors and arch length can be used as reliable parameters for stature analysis.

4.
JBJS Case Connect ; 11(3)2021 08 16.
Article in English | MEDLINE | ID: mdl-34398853

ABSTRACT

CASE: A full-term neonate presented with right lower extremity ischemia at birth because of spontaneous thrombosis of the right common iliac artery. He was initially managed with supportive treatment, anticoagulation, and dressings; however, advanced gangrenous changes precluded salvage of the ischemic limb. A guillotine amputation was performed at day 15 of life, and the stump went on to heal well by secondary intention. CONCLUSION: Thromboembolic events occurring in infancy are well-recognized phenomena; however, it is far rarer to encounter a neonate born with "congenital gangrene". We discuss the etiology, approach to diagnosis, and treatment of this rare but devastating condition.


Subject(s)
Arterial Occlusive Diseases , Thrombosis , Amputation, Surgical , Humans , Infant, Newborn , Ischemia/diagnosis , Ischemia/etiology , Ischemia/surgery , Lower Extremity , Male , Thrombosis/complications , Thrombosis/diagnostic imaging , Thrombosis/surgery
6.
J Clin Orthop Trauma ; 15: 1-8, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33717909

ABSTRACT

The Masquelet Induced Membrane Technique (IMT) is one of the tools in the surgeon's armamentarium for the management of segmental bone defects. The first stage of the IMT includes the insertion of a cement spacer, which is typically fashioned by the free-hand technique. We propose a novel technique for preparation of the cement spacer using a split syringe barrel as a mould. This technique produces a uniformly cylindrical spacer with minimum cement spillage, while also minimizing thermal damage to the surrounding soft tissues. It is a simple and cost-effective method that can be adapted for use in any long bone in children.

7.
Indian J Orthop ; 55(1): 23-34, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33569096

ABSTRACT

BACKGROUND: Femoral neck fractures in children are rare injuries, occurring due to high-energy trauma. Due to the unique anatomy and blood supply of the proximal femur in growing children, these fractures are notorious for high rates of complications despite appropriate management. Classification of these fractures is according to the Delbet system, which not only guides management but also gives prognostic clues. Multiple fixation methods have been described and there is no consensus on what constitutes the best treatment. Osteonecrosis, non-union, coxa vara and premature physeal arrest are the most frequent complications. PURPOSE: To review the current knowledge, discuss controversial aspects, and provide suggestions for future research. METHODS: We have reviewed the literature on paediatric proximal femur fractures and have provided an evidence-based guide to the diagnosis and management of these injuries. Common complications have been elaborated and options for their prevention and/or management discussed. CONCLUSION: There is universal agreement that anatomic reduction and stable internal fixation, supplemented by spica immobilization in younger children, are essential to obtain good outcomes. The role of capsular decompression, choice and configuration of implant, and appropriate timing of surgery are aspects that continue to be debated. Multicenter prospective studies are necessary to standardize treatment of these challenging injuries.

8.
Indian J Orthop ; 55(1): 100-108, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33569103

ABSTRACT

BACKGROUND: Modified Dunn procedure has become popular for the treatment of severe cases of slipped capital femoral epiphysis (SCFE). We assessed the outcomes in a consecutive series of thirty Indian adolescents treated by the modified Dunn procedure. MATERIALS AND METHODS: All patients treated by the modified Dunn procedure by a single senior Paediatric Orthopaedic surgeon over six years were retrospectively reviewed. Only moderate and severe slips undergoing modified Dunn procedure were included. Clinical records and radiographs were reviewed to obtain demographic information; to classify the slips by duration of symptoms, severity and physeal stability; and to assess the outcomes by Harris Hip Score, radiological changes and rate of complications. RESULTS: Thirty consecutive hips with 19 stable and 11 unstable slips were included. Mean age was 13.05 years, 25 boys and 5 girls; six were acute slips, six chronic and eighteen acute-on-chronic. There were 20 moderate and 10 severe slips. Slip angle correction was on average 43.63° ± 8.42° (p < 0.001). At a mean follow-up of 25.36 months, the slip angle averaged 9.9° ± 3.78°, and alpha angle was 33.63° ± 4.14. The average Harris Hip Score was 81.833 ± 7.12 points, with six excellent, 17 good, six fair and one poor result. Osteonecrosis occurred in two hips (6.6%). One hip had post-operative subluxation which was corrected. CONCLUSION: This study adds to the evidence that the modified Dunn procedure is safe, reliable and reproducible. It should be the first choice for the treatment of moderate and severe SCFE.

9.
Indian J Orthop ; 55(1): 147-157, 2021 Feb.
Article in English | MEDLINE | ID: mdl-33569109

ABSTRACT

PURPOSE: Traditional infant swaddling or binding with hips and knees extended is a known risk factor for Developmental Dysplasia of the Hip (DDH), while 'hip-safe swaddling' with hips and knees flexed is believed to eliminate this risk. We conducted a survey to determine the prevalent practices for infant swaddling in India; why mothers practice swaddling and who teaches them; and whether Paediatricians, nurses and caregivers are aware of hip-safe swaddling. METHODS: Anonymous one-time surveys were conducted in three groups-Paediatricians, Nurses and caregivers - at a tertiary-care, urban based, paediatric and maternity hospital. RESULTS: Forty-five paediatricians, 219 nurses and 100 caregivers were surveyed. Ninety percent caregivers practiced traditional swaddling, for on average 10.2 hours a day, starting soon after birth, up to 4.2 months of life. Traditional swaddling was advocated by 99% nurses and 53% Paediatricians. Reasons for swaddling included sleep, warmth and the misbelief that the child's legs would remain bowed if not bound straight; contrarily few mothers (8%) avoided swaddling out of superstition. Mothers learnt swaddling mainly from relatives (94%) and nurses (64%). Most nurses (70%) had learnt the practice during nursing training. Only 6.6% Paediatricians, 4% caregivers and 0% nurses were aware of 'hip-safe swaddling'. CONCLUSION: Traditional swaddling of infants is a practice deeply rooted in India, born out of misbeliefs, and propagated by lack of awareness. Training in hip-safe swaddling targeted at nurses and Paediatricians would be an effective initial step in creating awareness among mothers and changing their practices.

10.
Indian J Orthop ; 54(Suppl 2): 403-407, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33194111

ABSTRACT

The chondral shear fracture is a rare sub-type of pediatric capitellum fractures, in which the fractured fragment is almost entirely cartilaginous. Since the fractured fragment is not visible on plain radiographs, these injuries are often missed on initial presentation resulting in delayed diagnosis and poor outcomes. To our knowledge, only eight such cases have been reported in the past. We report the cases of two adolescents with chondral shear fractures of the capitellum, that were initially missed. They both presented to us several months following trauma, with pain and restricted elbow range of motion. They were treated by excision of the intra-articular loose fragment, and one child also required radial head excision due to advanced radio-capitellar arthritis. Both had good outcomes at 1 year follow-up. We provide a review of literature on this injury and emphasize the need for having a high index of suspicion when dealing with elbow trauma in adolescents, so as to avoid missing this rare, but distinct fracture pattern.

11.
J Int Soc Prev Community Dent ; 9(3): 256-262, 2019.
Article in English | MEDLINE | ID: mdl-31198698

ABSTRACT

AIMS AND OBJECTIVES: To evaluate the styloid process (SP) and its anatomical variations on digital panoramic radiographs from the database and also to synthesize the current evidence of literature on SP and anatomical variations along with the systematic review of the relevant studies after making the necessary exclusions. MATERIALS AND METHODS: A total of 500 panoramic radiographs from the database were examined and evaluated for the length of SP using OmniVue software and for the calcification patterns which were categorized into three types as described by Langlais. The obtained values were analyzed using t-test and Chi-square tests with a significance level of 0.005 and SPSS version 20.0 software. RESULTS AND CONCLUSION: The mean length of the SP in females was found to be 3.7 cm on the right side and 3.8 cm on the left side. The mean length of the SP in males was found to be 3.4 cm on the right side and 3.3 cm on the left side with significant difference between the genders with the P values of 0.0002 and 0.0001, respectively. The length of the SP was significantly longer in females than in males. Type I was the most common SP and was more prevalent in females. The results of the present study along with the wide range of reported incidence of anatomical variations in the form of elongation from the literature extracted through the systematic review suggests the need to reevaluate the range of the normal length of the SP.

12.
Fish Shellfish Immunol ; 62: 265-275, 2017 Mar.
Article in English | MEDLINE | ID: mdl-28119144

ABSTRACT

LGP2 (laboratory of genetics and physiology 2) is an important member of the retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs), which plays a significant role in antiviral innate immunity. In this study, we have cloned the full-length cDNA sequence of LGP2 from Asian seabass, Lates calcarifer (AsLGP2). The complete AsLGP2 cDNA sequence consisted of 2586 nucleotides encoding a putative protein of 681 amino acids with a molecular mass of 77.6 kDa. From the AsLGP2 protein, four different conserved domains were predicted: a DExDc (DEAD/DEAH box helicase domain), a bacterial type III restriction enzyme domain (RES III), a HELICc (Helicase superfamily c-terminal domain and a RIG-I_C-RD (RIG-I C-terminal regulatory domain). The transcript of AsLGP2 could be detected in all the 11 tissues tested in healthy animals with high expression noticed in tissues facing external environment such as gill, hindgut and skin. The ontogenic expression profile of AsLGP2 implies a possible maternal transfer of this gene as it has been detected in all early embryonic developmental stages along with unfertilized eggs. Viral analogue, poly I:C, injection resulted in rapid up-regulated expression in different tissues with the highest modulation of expression observed in kidney followed by liver and gill. A rapid response of AsLGP2 expression was also observed in the different tissues of Vibrio alginolyticus-injected L. calcarifer, while significant change in expression was noticed following Staphylococcus aureus infection. Similarly, exposure to different pathogen-mimicking microbial analogues such as poly I:C, LPS and PGN resulted in enhanced expression of AsLGP2 in SISK cell-line. Taking together, these observations suggest that AsLGP2 can act as both antiviral and antibacterial cytosolic receptor and may play a significant role in embryonic and larval development in marine euryhaline teleosts like Asian seabass.


Subject(s)
Fish Diseases/genetics , Fish Proteins/genetics , Perciformes , RNA Helicases/genetics , Staphylococcal Infections/veterinary , Vibrio Infections/veterinary , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , DNA, Complementary/genetics , DNA, Complementary/metabolism , Fish Diseases/immunology , Fish Diseases/microbiology , Fish Proteins/chemistry , Fish Proteins/metabolism , Pathogen-Associated Molecular Pattern Molecules , Phylogeny , RNA Helicases/chemistry , RNA Helicases/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Alignment/veterinary , Staphylococcal Infections/genetics , Staphylococcal Infections/immunology , Staphylococcal Infections/microbiology , Staphylococcus aureus , Tissue Distribution , Vibrio Infections/genetics , Vibrio Infections/immunology , Vibrio Infections/microbiology , Vibrio alginolyticus
13.
J Aerosol Med Pulm Drug Deliv ; 30(1): 1-13, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27537608

ABSTRACT

This article reports on discussions at the 2015 workshop cosponsored by the International Pharmaceutical Aerosol Consortium on Regulation and Science (IPAC-RS) and the International Society for Aerosols in Medicine (ISAM), entitled "Regulatory and Patient Considerations for Inhalation Device Design, Development and Use." Key topics addressed at the workshop and presented here include patient-focused device design for orally inhaled products (OIPs), instructions for use (IFU), human factors, regulatory considerations in the United States and Europe, development of generic inhalers, quality-by-design, and change management of OIP devices. Workshop participants also identified several areas for further consideration and emphasized the need for increased focus on the patient to create therapeutic products (inclusive of device design, IFU, education, training) that support adherence with an individual patient's treatment regimen. Advances in patient-centric product development will require engagement and collaboration by industry, regulators, patients, physicians, and other stakeholders. The article includes summaries of presented talks as well as of panel and audience discussions.


Subject(s)
Equipment Design , Medical Device Legislation , Nebulizers and Vaporizers , Administration, Inhalation , Aerosols , Europe , Humans , Pharmaceutical Preparations/administration & dosage , United States
14.
Fish Shellfish Immunol ; 55: 602-12, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27346158

ABSTRACT

Nod like receptors (NLRs) are a large group of cytoplasmic PRRs believed to play an important role in bacterial recognition in higher vertebrates. In this study, a novel Nod like receptor C3 (AsNLRC3) has been identified, cloned and characterised from Asian seabass, Lates calcarifer. The full-length AsNLRC3 transcript composed of a 4142 bp nucleic acid sequence encode for a protein of 1134 deduced amino acids. Three signature domains identified are conserved NACHT-domain, C-terminal LLR domain and N-terminal CARD effector domain. From the domain architecture and phylogenetic analysis, it was quite evident that AsNLRC3 is different from the NLR subfamily C of other teleosts. AsNLRC3 expressed in all the 11 tissues tested but highly expressed in tissues facing external environment such as gill, hindgut and midgut. The ontogenic expression profile of this receptor showed constitutive expression throughout the embryonic and larval developmental stages, which could be an innate immune strategy against different marine pathogens for larval survival. Infection with Vibrio alginolyticus and poly I:C induction showed an alteration of expression pattern in different tissues but did not show significant alteration in expression with Staphylococcus aureus infection. In vitro study in Asian seabass kidney cell line (SISK) stimulated with different ligands such as LPS, PGN and poly I:C showed considerable up-regulation at some of the time-points tested. These results suggest that AsNLRC3 can be a pivotal cytosolic innate immune receptor for recognizing wide array of pathogens in a euryhaline teleost model like Asian seabass in diverse environmental conditions.


Subject(s)
Fish Diseases/genetics , Fish Proteins/genetics , NLR Proteins/genetics , Perciformes , Poly I-C/pharmacology , Staphylococcal Infections/veterinary , Vibrio Infections/veterinary , Amino Acid Sequence , Animals , Base Sequence , Caspase Activation and Recruitment Domain , Cloning, Molecular , DNA, Complementary/genetics , DNA, Complementary/metabolism , Fish Diseases/immunology , Fish Diseases/microbiology , Fish Proteins/chemistry , Fish Proteins/metabolism , NLR Proteins/chemistry , NLR Proteins/metabolism , Phylogeny , Sequence Alignment/veterinary , Staphylococcal Infections/genetics , Staphylococcal Infections/immunology , Staphylococcal Infections/microbiology , Staphylococcus aureus/physiology , Vibrio Infections/genetics , Vibrio Infections/immunology , Vibrio Infections/microbiology , Vibrio alginolyticus/physiology
15.
Ayu ; 36(4): 397-403, 2015.
Article in English | MEDLINE | ID: mdl-27833367

ABSTRACT

INTRODUCTION: India is one of the countries with high prevalence of anemia during pregnancy. Anemia in pregnancy is multifactorial. Iron deficiency anemia is the most common conditions in a pregnant woman. As per ayurvedic classics, this condition occurs due to improper Rasa Dhatu in mother and continuously increasing fetal demands and is considered as Rasa Pradoshajavikara. A large number of Lauha preparations have been used widely for centuries to cure Anemia. AIM: To evaluate efficacy of Punarnava Mandura and Dhatri Lauha on Garbhini Pandu. MATERIALS AND METHODS: A total 24 pregnant women with symptoms of Garbhini Pandu were randomly divided into two groups (A and B). In Group A (n = 15) Punarnava Mandura, two tablets (each of 500 mg) thrice a day with one cup (100 ml) of buttermilk and in Group B (n = 9) Dhatri Lauha, two tablets (each of 500 mg) thrice a day with luke warm water were administered for 90 days. The assessment was done with subjective parameters such as pallor, general weakness, dyspnea, etc., and objective parameters such as hematological parameters. Results were statistically analyzed using Student's t-test. RESULTS: The results revealed that overall clinical improvement was better in Group A when compared to Group B. Hemoglobin was increased in patients of Group A, which was statistically significant. No adverse drug reaction was observed during the treatment period. CONCLUSION: Punarnava Mandura is more effective on Garbhini Pandu in comparison to Dhatri Lauha.

16.
Gene ; 557(2): 172-81, 2015 Feb 25.
Article in English | MEDLINE | ID: mdl-25523097

ABSTRACT

Innate immune system recognises pathogen-associated molecular patterns (PAMPs) by limited number of germline encoded and non-clonally developed pathogen recognition receptors (PRRs). Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) are important cytosolic PRRs for sensing viral RNAs. The receptor encoded by melanoma differentiation associated gene 5 (MDA5), an RLR, recognises viral RNA and enhances antiviral response in host cells. The full-length MDA5 cDNA in Etroplus suratensis was cloned and found to have 3673 nucleotides encoding a polypeptide of 978 amino acids. The deduced amino acid sequence contains four main structural domains: two CARD domains in the N-terminal region, a DExDc (DEAH/DEAD box helicase domain), HELICc (C-terminal helicase) domain and a C-terminal regulatory domain (RD). Phylogenetic analysis revealed a close relationship of E. suratensis MDA5 (EsMDA5) with MDA5 of Neolamprologus brichardi and Oreochromis niloticus, both belonging to Cichlidae family. EsMDA5 transcripts were ubiquitously expressed in all the 12 tissues tested in healthy fish. Although, transcript level was found to be the highest in muscle, high expression was also detected in the spleen, head kidney and hindgut. In poly I:C-injected fish, EsMDA5 transcripts showed peak expression in the spleen, intestine and heart at 12h post-injection (hpi). However, in gill and kidney tissues, maximum up-regulation of EsMDA5 was observed at 6 and 48 hpi, respectively. Further, liver tissue showed an increasing trend in expression profile from 6 to 48 hpi. Interferon promoter stimulator-1 (IPS-1) gene, an adaptor triggering RIG-I- and MDA5-mediated type I interferon induction, also showed up-regulated expression at initial time-points in poly I:C-injected E. suratensis. The constitutive expression and up-regulation of EsMDA5 and the IPS-1 genes in different tissues indicate that EsMDA5 may play an important role in sensing viral PAMPs in conjunction with IPS-1.


Subject(s)
Cichlids/genetics , DEAD-box RNA Helicases/metabolism , Fish Proteins/metabolism , Amino Acid Sequence , Animals , Base Sequence , Cichlids/metabolism , Cloning, Molecular , Conserved Sequence , DEAD-box RNA Helicases/genetics , Fish Proteins/genetics , Gene Expression , Molecular Sequence Data , Organ Specificity , Phylogeny , Sequence Analysis, DNA
17.
Fish Shellfish Immunol ; 41(2): 441-54, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25266891

ABSTRACT

The Toll-pathway plays key roles in regulating the innate immune response in invertebrates. Myeloid differentiation factor 88 (MyD88) and Tumour necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) are key molecules in this signalling pathway. To investigate the role of Toll-pathway in innate immune response of shrimp, Penaeus monodon, MyD88 (PmMyD88) and TRAF6 (PmTRAF6) were identified and characterised. PmMyD88 cDNA is 1716 bp long with an open reading frame (ORF) of 1449 bp encoding a putative protein of 482 amino acids, with a death domain, a TIR domain and C-terminal extension domain. PmTRAF6 cDNA is 2563 bp long with an ORF of 1785 bp (594 amino acids) with an N-terminal RING-type zinc finger domain, two TRAF-type zinc finger domains, a coiled region and a MATH domain. In healthy shrimp, PmMyD88, PmTRAF6 and PmToll were detected in 15 tissues with the highest expression in midgut, eyestalk and lymphoid organ, respectively. Responses of these genes to WSSV in experimentally-infected P. monodon as well as in cultured haemocytes and also effect of poly I:C on the gene expression in vitro was investigated at six time-points in seven tissues. PmToll showed significant up-regulation at all time-points of infection in six tissues and until 24 h post-infection in vitro. However, poly I:C-induced haemocytes showed up-regulation of the gene until 48 h post-exposure. WSSV caused significant up-regulation of PmMyD88 in most of the tissues tested. The virus challenge as well as poly I:C induction in vitro also resulted in significant up-regulation of the gene. Up-regulated expression of PmTRAF6 was detected in haemocytes and lymphoid organ at late stage of infection. In vitro virus challenge showed significant up-regulation of PmTRAF6 at almost all time-points whereas no significant change in the expression was observed on poly I:C induction. The responses of these key genes, observed in the present study, suggest that Toll-pathway as a whole may play a crucial role in the immune response against viruses in shrimp.


Subject(s)
Gene Expression Regulation/immunology , Immunity, Innate/genetics , Penaeidae/immunology , Penaeidae/virology , Signal Transduction/immunology , White spot syndrome virus 1/immunology , Animals , Cloning, Molecular , Computational Biology , DNA Primers/genetics , Gene Expression Profiling , Myeloid Differentiation Factor 88/metabolism , Real-Time Polymerase Chain Reaction , Signal Transduction/genetics , TNF Receptor-Associated Factor 6/metabolism , Toll-Like Receptors/metabolism
18.
Mol Biol Rep ; 41(10): 6865-77, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25012916

ABSTRACT

Toll-like receptors are sentinels of innate immune system, which recognise pathogen-associated molecular patterns, and subsequently activate production of antimicrobial peptides to contain the infection. In the present study, we cloned and characterised a Toll gene from Scylla serrata (SsToll) encoding 1005 amino acids with typical Toll-like receptor domain topology. Phylogenetic analysis revealed that it belongs to insect-type invertebrate Toll family showing 100 % identity with Scylla paramamosain (SpToll). The expression pattern of mRNA in different tissues indicated that SsToll is constitutively expressed in all the tissues examined, with varying expression levels. The expression was also detected in all the life-stages (egg, zoea stages 1-5, megalopa and crab instar) with the highest level observed in zoea 2. In-vitro studies using crab haemocyte culture demonstrated that SsToll transcripts are distinctly modulated in response to ligands such as peptidoglycan and lipopolysaccharide at all time-points. A significant change in SsToll expression was also noticed in haemocytes exposed to poly I:C (3-9 h). On the contrary, the transcription level of SsToll in response to white spot syndrome virus (WSSV) challenge was noticeably different. The change in expression in vitro was not significant at early time-points until 3 h; the transcripts showed a significant up-regulation commencing from 6 h, but not beyond 12 h. However, in vivo expression was unaffected at early time-points of WSSV challenge (until 12 h) and a gradual up-regulation was detected at 24 h. In-vivo challenge with Vibrio parahaemolyticus resulted in delayed up-regulation of the gene. The results obtained in the present study suggest that SsToll might be involved in the innate immunity of mud crab.


Subject(s)
Brachyura/genetics , Toll-Like Receptors/genetics , Amino Acid Sequence , Animals , Base Sequence , Brachyura/metabolism , Brachyura/microbiology , Brachyura/virology , Cloning, Molecular , Gene Expression Profiling , Gene Expression Regulation , Host-Pathogen Interactions/genetics , Ligands , Molecular Sequence Data , Organ Specificity/genetics , Phylogeny , Sequence Alignment , Sequence Analysis, DNA , Toll-Like Receptors/metabolism
19.
In Vitro Cell Dev Biol Anim ; 50(5): 406-16, 2014.
Article in English | MEDLINE | ID: mdl-24357036

ABSTRACT

Primary cell cultures were developed from haemocytes and testis of Scylla serrata. Haemocytes were collected from live animals and cultured in double-strength L-15 medium (2× L-15) prepared in crab saline, supplemented with 5% foetal bovine serum and antibiotic-antimycotic solution (penicillin 100 U/mL, streptomycin 100 µg/mL and amphotericin B 0.25 µg/mL) with osmolality adjusted to 894 mOsm/kg. The haemocytes adhered within 2 h after seeding and showed proliferation up to 72 h. The disaggregated testis tissue fragments were seeded in 3× L-15 supplemented with non-essential amino acid mixture, lipid concentrate and antibiotic-antimycotic solution, with osmolality adjusted to 1,035 mOsm/kg with crab saline. Cells from the testis could be subcultured and maintained up to 21 d as suspension culture. Different dilutions of white spot syndrome virus (WSSV) inoculum (known virus copy number) prepared from infected Penaeus monodon were inoculated in the cultured cells, and the cytopathic effects like detachment, rounding of cells and clear areas of depleted cells were observed after 48 h in haemocyte cultures. However, WSSV-exposed testis cells did not show any obvious change until 72 h post-infection. WSSV was detected in both haemocyte and testis cultures at different time-points of infection by conventional and real-time PCR using WSSV-specific primers. The transcripts of WSSV were found to be much higher in haemocytes than in testis culture. The virus harvested from the cultured haemocytes after three passages could infect healthy P. monodon. The present study showed that mud crab haemocyte culture can support WSSV replication, and it can be used as an in vitro tool for WSSV replication.


Subject(s)
Brachyura/cytology , In Vitro Techniques , Primary Cell Culture/methods , Animals , Brachyura/virology , Virus Replication , White spot syndrome virus 1/genetics , White spot syndrome virus 1/growth & development
20.
J Invertebr Pathol ; 111(1): 86-9, 2012 Sep 15.
Article in English | MEDLINE | ID: mdl-22705108

ABSTRACT

Monoclonal antibodies (MAbs) to hemocytes of mud crab, Scylla serrata, were produced by immunizing mice with formalin-fixed hemocytes. Of the six MAbs produced, two (MAb 1D11 and MAb 1A2) reacted specifically with hemocyte proteins in western blot. MAb 1A2 showed strong immunofluorescent reaction with granular hemocytes and a weak reaction with semigranular cells. However, hyaline cells did not show any reaction. The MAbs also showed strong cross-reactivity with the hemocytes of different crab species but not with other crustaceans. The MAbs produced can be used as a marker for granular hemocytes of crabs.


Subject(s)
Antibodies, Monoclonal/immunology , Brachyura/immunology , Hemocytes/immunology , Hemolymph/immunology , Animals , Immunoglobulin Isotypes , Mice
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