Design, synthesis and ex-vivo release studies of colon-specific polyphosphazene-anticancer drug conjugates.

Colon-specific azo based polyphosphazene-anticancer drug conjugates (11-18) have been synthesized and evaluated by ex-vivo release studies. The prepared polyphosphazene drug conjugates (11-18) are stable in acidic (pH=1.2) buffer which showed that these polymer drug conjugates are protected from acidic environment which is the primary requirement of colon specific targeted drug delivery. The ex-vivo release profiles of polyphosphazene drug conjugates (11-18) have been performed in the presence as well as in the absence of rat cecal content. The results showed that more than 89% of parent drugs (methotrexate and gemcitabine) are released from polymeric backbone of polyphosphazene drug conjugates (14 and 18) having n-butanol (lipophilic moiety). The in-vitro cytotoxicity assay has also been performed which clearly indicated that these polymeric drug conjugates are active against human colorectal cancer cell lines (HT-29 and COLO 320 DM). The drug release kinetic study demonstrated that Higuchi's equation is found to be best fitted equation which showed that release of drug from polymeric backbone as square root of time dependent process based on non-fickian diffusion. Therefore, the synthesized polyphosphazene azo based drug conjugates of methotrexate and gemcitabine are the potential candidates for colon targeted drug delivery system with minimal undesirable side effects.

The expression of IL6 and 21 in crossbred calves upregulated by inactivated trivalent FMD vaccine.

Foot and mouth disease (FMD) is an economically important disease and a whole-virus inactivated trivalent virus vaccine is the mainstay for controlling the disease in India. The protective humoral immune response to FMD vaccination is a complex, but, tightly regulated process mediated by the interplay of interleukins (IL). Based on the specific role of IL6 and 21 in adaptive immune response, we hypothesized that inactivated trivalent FMD vaccine would stimulate IL6 and 21 expression in the circulating lymphocytes. The expressions of IL6 and 21 were assayed on 0, 28, 60, 90, and 120 d post-vaccination (DPV) by quantitative PCR (qPCR) with simultaneous assessment of FMDV antibody titer by liquid phase blocking ELISA. The results revealed that the peak expression of IL6 and 21 was on DPV 28 which correlated well with the FMDV antibody titer and plummeted to the prevaccination titer level by 60 DPV. As IL21 is the final effector of antibody production as compared to IL6, we investigated the expression of IL21 in calves that had protective titer (>1.8) with the unprotected group (<1.8). Expression of IL21 on 28 DPV was numerically higher in the protected than that of the unprotected group of calves.

Use of knotless barbed sutures for closure of intraoral incisions for maxillofacial trauma: a randomised controlled trial.

The objective of this study was to assess the efficacy of knotless barbed sutures in intraoral wound closure for maxillofacial trauma in comparison with conventional (vicryl) sutures. This was a randomised controlled clinical trial involving 40 patients with isolated mandibular angle fractures who required intraoral incisions for open reduction and internal fixation (ORIF). The sample was randomised into the study group (20 patients) and control group (20 patients). Following fracture fixation by a standardised surgical protocol, the wound closure was done with bidirectional knotless barbed suture and vicryl for the study and control groups, respectively. The wounds were closed in layers (periosteum and mucosa). All operations were performed by a single surgeon. Outcome parameters measured were intraoperative wound closure time and wound healing using 'Landry's wound healing index' on the first, third, and seventh postoperative days. Statistically significant difference in suturing time was noted between the study and control group (p value <0.001). The study group demonstrated a mean (SD) suturing time of 9.46 (2.01) minutes, compared with the 17.61 (2.57) minutes in the control group. Wound healing was found to be better and statistically significant in the study group than the control group (p value<0.001). Knotless barbed suture is a promising alternative to vicryl for intraoral wound closure.

Marsupialization: a conservative approach for treating dentigerous cyst in children in the mixed dentition.

Dentigerous cysts are usually encountered in the practice of pediatric dentistry. The treatment modalities range from marsupialization to enucleation of the lesion and are based on the involvement of the lesion with the adjacent structures. However, loss of a permanent tooth in the management of a dentigerous cyst can be devastating to a child who has already a congenitally missing tooth. The first case describes the technique of marsupialization in which we extracted the grossly carious deciduous 1st molar and created a window through the extracted socket to decompress the lesion. In this case the 2nd premolars were congenitally missing on both sides of the mandible for which we had not gone for enucleation of the dentigerous cyst along with the developing 1st premolar. The second case is a developmental type of a big dentigerous cyst where marsupialization was followed by enucleation of the cystic lining but without removal of the affected tooth. Both the teeth erupted in the oral cavity.

Poly(methylmethacrylate) grafted chitosan: An efficient adsorbent for anionic azo dyes.

Present study reports on peroxydisulfate/ascorbic acid initiated synthesis of Chitosan-graft-poly(methylmethacrylate) (Ch-g-PMMA) and its characterization by FTIR, XRD and (13)C NMR. The copolymer remained water insoluble even under highly acidic conditions and was evaluated to be an efficient adsorbent for the three anionic azo dyes (Procion Yellow MX, Remazol Brilliant Violet and Reactive Blue H5G) over a wide pH range of 4-10 being most at pH 7. The adsorbent was also found efficient in decolorizing the textile industry wastewater and was much more efficient than the parent chitosan. Equilibrium sorption experiments were carried out at different pH and initial dye concentration values. The experimental equilibrium data for each adsorbent-dye system were successfully fitted to the Langmuir and Freundlich sorption isotherms. Based on Langmuir model Q(max) for yellow, violet and blue dyes was 250, 357 and 178, respectively. Thermodynamic parameters of the adsorption processes such as DeltaG degrees , DeltaH degrees , and DeltaS degrees were calculated. The negative values of free energy reflected the spontaneous nature of adsorption. The adsorption kinetic data of all the three dyes could be well represented by pseudo-second-order model with the correlation coefficients (R(2)) being 0.9922, 0.9997 and 0.9862, for direct yellow, reactive violet and blue dye, respectively with rate constants 0.91 x 10(-4), 1.82 x 10(-4) and 1.05 x 10(-4) g mg(-1)min(-1), respectively. At pH 7, parent chitosan also showed pseudo-second-order kinetics. The temperature dependence of dye uptake and the pseudo-second-order kinetics of the adsorption indicated that chemisorption is the rate-limiting step that controls the process.

Biological approach in repair of partially inflamed dental pulp using second-generation platelet-rich fibrin and mineral trioxide aggregate as a pulp medicament in primary molars.

AIM: The purpose of this study was to evaluate and compare clinically and radiographically PRF, MTA and Ca(OH)2 as a pulp dressing material in primary molars. MATERIAL AND METHODS: The study was conducted on 30 children between 6-9 years of age. Pulpotomy was performed using MTA(Gp I), PRF + MTA(Gp II) and PRF +Ca(OH)2(Group III). The teeth were evaluated clinically and radiographically after 1,3 and 6 months and were subjected to statistics analysis. RESULTS: After 6 months follow-up the overall success rate was found to be highest in Gp I with success rate of 97% followed by Gp II 95% and Gp III 78.3%. The results were found to be statistically insignificant (P ≤ 0.05). CONCLUSION: All materials used in the present study were equally effective as the success rates are statistically insignificant.

A cross-sectional study to assess the prevalence of early childhood caries and associated risk factors in preschool children in district Mandi, Himachal Pradesh.

AIM: The aim of this cross-sectional study is to investigate the prevalence and related risk factors of early childhood caries (ECC) in preschool children of district Mandi, Himachal Pradesh, India. MATERIALS AND METHODS: A total sample of 2859 children aged between 2 and 6 years were selected from various parts of district Mandi. The status of dental caries and decayed missing and filled teeth (DMFT) score was recorded. RESULTS: ECC increased significantly with age. Boys had significantly higher caries prevalence and mean DMFT score as compared to girls. Children who belonged to low socioeconomic group showed higher caries prevalence and mean DMFT score. Children aged 3-6 years who brushed their teeth twice daily had the least prevalence of dental caries as well as DMFT score when compared to children with one time and no brushing. CONCLUSION: The early identification of poor oral hygiene and improper feeding habits should be considered in preventive health promotion in low socioeconomic communities of district Mandi, Himachal Pradesh.

Spectroscopic investigations of carious tooth decay.

We report on the elemental composition of healthy and infected part of human tooth using laser induced breakdown spectroscopy (LIBS). We have used prominent constituent transitions in laser-excited tooth to diagnose the state of the tooth. A nanosecond laser pulse (355nm, 5ns) was used as an ablating pulse and the sodium (3s2S-3p2P) at 588.99 and (3s2S-3p2P) at 589.99nm, strontium (5s21S-1s5P) at 460.55nm, and calcium (3d3D-4f 3F0) at 452.55nm transitions for spectroscopic analysis. The spectroscopic observations in conjunction with discriminate analysis showed that calcium attached to the hydroxyapatite structure of the tooth was affected severely at the infected part of the tooth. The position-time plots generated from two-dimensional (2D) images conclusively showed a decrease in calcium concentration in the infected region of the irradiated tooth. Using the technique, we could distinguish between the healthy and carious parts of the tooth with significant accuracy.

Foot-and-mouth disease virus transmission dynamics and persistence in a herd of vaccinated dairy cattle in India.

Foot-and-mouth disease (FMD) is an important transboundary disease with substantial economic impacts. Although between-herd transmission of the disease has been well studied, studies focusing on within-herd transmission using farm-level outbreak data are rare. The aim of this study was to estimate parameters associated with within-herd transmission, host physiological factors and FMD virus (FMDV) persistence using data collected from an outbreak that occurred at a large, organized dairy farm in India. Of 1,836 regularly vaccinated, adult dairy cattle, 222 had clinical signs of FMD over a 39-day period. Assuming homogenous mixing, a frequency-dependent compartmental model of disease transmission was built. The transmission coefficient and basic reproductive number were estimated to be between 16.2-18.4 and 67-88, respectively. Non-pregnant animals were more likely to manifest clinical signs of FMD as compared to pregnant cattle. Based on oropharyngeal fluid (probang) sampling and FMDV-specific RT-PCR, four of 36 longitudinally sampled animals (14%) were persistently infected carriers 10.5 months post-outbreak. There was no statistical difference between subclinical and clinically infected animals in the duration of the carrier state. However, prevalence of NSP-ELISA antibodies differed significantly between subclinical and clinically infected animals 12 months after the outbreak with 83% seroprevalence amongst clinically infected cattle compared to 69% of subclinical animals. This study further elucidates within-herd FMD transmission dynamics during the acute-phase and characterizes duration of FMDV persistence and seroprevalence of FMD under natural conditions in an endemic setting.

Quantitative characteristics of the foot-and-mouth disease carrier state under natural conditions in India.

The goal of this study was to characterize the properties and duration of the foot-and-mouth disease (FMD) carrier state and associated serological responses subsequent to vaccination and naturally occurring infection at two farms in northern India. Despite previous vaccination of cattle in these herds, clinical signs of FMD occurred in October 2013 within a subset of animals at the farms containing juvenile-yearling heifers and steers (Farm A) and adult dairy cattle (Farm B). Subsequent to the outbreak, FMD virus (FMDV) asymptomatic carriers were identified in both herds by seroreactivity to FMDV non-structural proteins and detection of FMDV genomic RNA in oropharyngeal fluid. Carriers' seroreactivity and FMDV genome detection status were subsequently monitored monthly for 23 months. The mean extinction time of the carrier state was 13.1 ± 0.2 months, with extinction having occurred significantly faster amongst adult dairy cattle at Farm B compared to younger animals at Farm A. The rate of decrease in the proportion of carrier animals was calculated to be 0.07 per month. Seroprevalence against FMDV non-structural proteins decreased over the course of the study period, but was found to increase transiently following repeated vaccinations. These data provide novel insights into viral and host factors associated with the FMDV carrier state under natural conditions. The findings reported herein may be relevant to field veterinarians and governmental regulatory entities engaged in FMD response and control measures.

Polypropylene mesh seeded with fibroblasts: A new approach for the repair of abdominal wall defects in rats.

PURPOSE: The purpose of study was to develop bioengineered scaffolds by seeding primary mouse embryo fibroblast cells (p-MEF) on polypropylene mesh and to test its efficacy for the repair of abdominal wall defects in rats. METHODS: The study was conducted on 18 clinically healthy adult Wistar rats of either sex. The animals were randomly divided into two equal groups having nine animals in each group. In both the groups a 20mm×20mm size full thickness muscle defect was created under xylazine and ketamine anesthesia in the mid-ventral abdominal wall. In group I the defect was repaired with polypropylene mesh alone and in group II it was repaired with p-MEF seeded polypropylene mesh. Matrices were implanted by synthetic absorbable suture material (polyglycolic acid) in continuous suture pattern. The efficacy of the bio-engineered matrices in the reconstruction of full thickness abdominal wall defects was evaluated on the basis of macro and histopathological observations. RESULTS: Macroscopic observations revealed that adhesions with skin and abdominal viscera were minimum in group II as compared to group I. Histopathological observations confirmed better fibroplasia and collagen fiber arrangement in group II. No recurrence of hernia was found in both the groups. CONCLUSION: Hernias are effectively repaired by implanting polypropylene mesh. However, this work demonstrates that in vitro seeding of mesh with fibroblasts resulted in earlier subsidization of pain, angiogenesis and deposition of collagen, increased thickness of matrices with lesser adhesions with underlying viscera. On the basis of the results p-MEF seeded mesh was better than non-seeded mesh for repair of abdominal wall defects in rats.

Three-dimensional structure of mare diferric lactoferrin at 2.6 A resolution.

Lactoferrin is a monomeric glycoprotein with a molecular mass of approximately 80 kDa. The three-dimensional structure of mare diferric lactoferrin (mlf) has been determined at 2.6 A resolution. The protein crystallizes in the space group P 212121with a=85.2 A, b=99.5 A, c=103.1 A with a solvent content of 55 % (v/v). The structure was solved by the molecular replacement method using human diferric lactoferrin as the model. The structure has been refined using XPLOR to a final R -factor of 0.194 for all data in the 15-2.6 A resolution range. The amino acid sequence of mlf was determined using a cDNA method. The final refined model comprises 5281 protein atoms, 2 Fe3+, 2 CO32-and 112 water molecules. The overall folding of mlf is similar to that of other proteins of the transferrin family. The protein folds into two globular lobes, N and C. The lobes are further divided into two domains, N1 and N2, and C1 and C2. The iron-binding cleft is situated between the domains in each lobe. The N lobe appears to be well ordered and is more stable than the C lobe in mlf unlike in other lactoferrins, where the C lobe is the more stable. The opening of the binding cleft in the N lobe of mlf is narrower than those in other proteins of the transferrin family. This is very unusual and is found only in mare lactoferrin. Apart from certain hydrophobic interactions at the mouth of the cleft, one salt-bridge (Lys301 . . . . . . . . Glu216) crosses between the two walls of the cleft. The two lobes are connected covalently by a three-turn alpha-helix involving residues 334-344. The N lobe displays a highly ordered structure with appreciably low temperature factors. The iron coordination is more symmetrical in the N lobe than in the C lobe. There are only 16 intermolecular hydrogen bonds in the structure of mlf.

GAPO syndrome in a child without dermal hyaline deposit.

A 5-year-old girl with GAPO syndrome from India lacked PAS-positive hyaline material in the skin biopsy from thigh and scalp. The role of this pathological change, earlier reported by Wajntal et al. [1990] in the pathogenesis of GAPO syndrome, needs to be reexamined.

The adhesion of glow--discharge polymers, Silastic and Parylene to implantable platinum electrodes: results of tensile pull tests after exposure to isotonic sodium chloride.

Thin organic coatings commonly are used for insulating microelectrodes and electronic packages designed for implant applications. The adherence of these coatings to the underlying substrates is a key parameter in their selection for various devices. Instron pull tests were performed on glow-discharge polymerized monomers, Parylene-N, medical-grade Silastic and various epoxies. The application of a thin coating of glow-discharge polymerized methane under a thicker Parylene-N coating improved the adhesion of the latter to the underlying substrate in isotonic sodium chloride solution and during accelerated testing conditions done by boiling.

Development of a radioimmunoassay procedure for aflatoxin B1 measurement.

A radioimmunoassaay (RIA) procedure to measure aflatoxin B(1) (AfB(1)) in agricultural commodities was developed. AfB(1) oxime derivative was synthesized, characterized, and used for preparation of (125)I-labeled AfB(1). Antiaflatoxin B(1) serum was raised in-house using AfB(1)-bovine serum albumin conjugate as immunogen. The assay system was optimized in the range of 0.2-5 ng/mL, using a liquid phase (PEG) as well as a solid phase (coated polystyrene beads) separation system. Inter-assay and intra-assay variations, recovery, and parallelism studies validated the assay. AfB(1) analysis was carried out in nearly 130 samples of different agricultural commodities. The correlation coefficient was determined using commercial ELISA and in-house-developed RIA methods.

A new syndrome of multiple joint dislocations with metaphyseal dysplasia.

A 15-day-old female child is presented with multiple joint dislocations who died during the neonatal period. She had additional features of metaphyseal dysplasia, deficient calcification of vault of the skull, growth retardation, a natal tooth, lymphoedema and facial dysmorphism. This may constitute a new syndrome of multiple joint dislocations with metaphyseal dysplasia.

Effect of vitamin A excess on oral armature in tadpoles of toad, Bufo melanostictus (Schneider) and frog, Rana cyanophlyctis (Schneider).

Tadpoles of B. melanostictus and R. cyanophlyctis were reared in vitamin A solution. Rearing of the tadpoles of B. melanostictus in solutions containing 15, 20 and 30 IU/ml vitamin A palmitate resulted in complete disappearance of the keratinized epidermal material over the jaws and horny labial teeth. However tadpoles of the frog R. cyanophlyctis, were not affected in this way.

In vitro control of fasciation in proliferating nucellar embryos of Mangifera indica L. var totapari red small for cloning.

Nucellar tissue contained in ovular halves of young fruits of Mangifera indica L. totapari red small, a dwarfing rootstock, differentiated fasciated embryonal structures in presence of 6-benzylaminopurine [BAP(0.15 mg l(-1))], 6-(gamma-gamma-dimethylallylamino) purine [2iP(0.15 mg l(-1))] and indole-3-acetic acid [(IAA(0.5 mg l(-1))] incorporated in the semisolid medium during 50-60 days. Due to embryonal fasciation, hardly 2-3 well-formed embryos could be obtained per culture of proliferating embryos. Of the 3 ethylene inhibitors [L-alpha-(2-aminoethoxyvinyl)-glycine-HCl (AVG), AgNO3 and salicylic acid (SA)] used, embryonal fasciation and necrosis of intervening tissue was completely controlled by 3-4 subcultures of fasciated mass of embryos under the influence of AVG (0.05 mg l(-1)) in presence of adenine sulphate [AdS (50 mg l(-1))] incorporated in the same medium. Almost synchronized development of isolated embryos, measuring ca 2 cm in length, was observed in a different medium used in liquid stationary state and supplemented, particularly with stress-producing substances [abscisic acid (ABA, 0.01 mg l(-1)); and polyethylene glycol (PEG, 100 mg l(-1))] besides certain other modifications. About 34% convertibility of processed embryos was obtained during a period of 90 days. The plantlets had well-developed roots along with laterals which were longer than leafy shoots. In vitro raised plants survived ex vitro for about 2 months.

Three-dimensional structure of a new form of mare lactoferrin in 70% PEG 400 at 3.8 A resolution.

Three-dimensional (3D) structure of a new form of diferric mare lactoferrin has been determined at 3.8 A resolution. The protein was crystallized in a space group P2(1)2(1)2(1) with a = 80.1 A, b = 103.7 A, c = 112.2 A with a solvent content of 57%. The structure was solved by molecular replacement method using the model of native mare lactoferrin. The structure has been refined using X-PLOR to a final R-factor of 22.6% for all the data in 15.0-3.8 A resolution range. The final refined model comprises 5281 protein atoms, 2Fe3+ and 2CO3(2-) ions. The protein folds into two globular N- and C-lobes. The two lobes are further divided into two domains N1, N2 in the N-lobe and C1, C2 in the C-lobe. The overall folding of the protein is similar to that observed for the native protein. The superposition of Calpha traces of native mare lactoferrin and the present structure gives an r.m.s shift of 0.7 A. There is a slight variation in the orientation of two lobes but the domain orientations in the present structure are identical to those observed in the native mare lactoferrin.

Association of BoLA DRB3 alleles with variability in immune response among the crossbred cattle vaccinated for foot-and-mouth disease (FMD).

Polymorphism of bovine leukocyte antigen (BoLA) DRB3 gene is being intensively investigated for potential association with economically important diseases of cattle. Accordingly, we investigated the association of DRB3 Exon 2 polymorphism as evidenced by the variation in the binding pockets with variability in immune response to inactivated trivalent (O, A and Asia1) foot and mouth disease virus (FMDV) vaccine in a closed population of crossbred cattle. Antibody titer of ≥ 1.8 was set as the cut off value to distinguish the protected (≥ 1.8) and unprotected (<1.8) animals. Eleven different alleles of over 3% frequency were detected in the population. We found that DRB3 alleles 0201, 0801 and 1501 always ranked high for protective immune response whereas alleles 0701, 1103 and 1101 consistently ranked low for unprotected immune response for all the three serotypes. Rank correlation of DRB3 alleles among the three serotypes was positive, high in magnitude and statistically significant (P<0.05). Logistic regression analysis revealed that odds of protection from the vaccine were highest for all the three serotypes if allele (∗)1501 was present and strengthened the results of allele ranking. Predicted amino acid substitution in the peptide binding pockets revealed that all the important sites had high Wu-Kabat index. Similarly, specific residues in pockets were crucial for immune response to FMD vaccine. There were specific substitutions in un-protected alleles such as absence of acidic amino acids substituted by basic amino acid at ß71, presence of non-polar cysteine or basic histidine at ß30 and presence of polar tyrosine at ß37. From the observations, we hypothesize that the substitutions lead to unique conformational changes in the protein products of the studied alleles that would associate with the protective or unprotective antibody response to FMDV vaccine. The knowledge has potential implications in future selection programs if integrated with the complete BoLA haplotype details and production traits of the herd.