Outcome Analysis Comparing Muscle and Fasciocutaneous Free Flaps for Heel Reconstruction: Meta-Analysis and Case Series.

Background Choosing the components of free flap (fasciocutaneous or muscle) is one of the crucial but controversial decisions in heel reconstruction. This meta-analysis aims to provide an up-to-date comparison of fasciocutaneous flaps (FCFs) and muscle flaps (MFs) for heel reconstruction and to ascertain if one flap has an advantage over the other. Methods Following the Preferred Reporting Item for Systematic Reviews and Meta-Analyses guidelines, a systematic literature review was performed identifying studies on heel reconstruction with FCF and MF. Primary outcomes were survival, time of ambulation, sensation, ulceration, gait, need for specialized footwear, revision procedures, and shear. Meta-analyses and Trial Sequential Analysis (TSA) were performed to estimate the pooled risk ratios (RRs) and standardized mean difference (SMD) with fixed effects and random effects models, respectively. Results Of 757 publications identified, 20 were reviewed including 255 patients with 263 free flaps. The meta-analysis showed no statistically significant difference between MF and FCF in terms of survival (RR, 1; 95% confidence interval [CI], 0.83, 1.21), gait abnormality (RR, 0.55; 95% CI, 0.19, 1.59), ulcerations (RR, 0.65; 95% CI, 0.27, 1.54), footwear modification (RR, 0.52; 95% CI, 0.26, 1.09), and revision procedures (RR, 1.67; 95% CI, 0.84, 3.32). FCF had superior perception of deep pressure (RR, 1.99; 95% CI, 1.32, 3.00), light touch, and pain (RR, 5.17; 95% CI, 2.02, 13.22) compared with MF. Time to full weight-bearing (SMD, -3.03; 95% CI, -4.25, -1.80) was longer for MF compared with FCF. TSA showed inconclusive results for comparison of the survival of flaps, gait assessment, and rates of ulceration. Conclusion Patients reconstructed with FCF had superior sensory recovery and early weight bearing on their reconstructed heels, hence faster return to daily activities compared with MFs. In terms of other outcomes such as footwear modification and revision procedure, both flaps had no statistically significant difference. The results were inconclusive regarding the survival of flaps, gait assessment, and rates of ulceration. Future studies are required to investigate the role of shear on the stability of the reconstructed heels.

Differential host responses of sugarcane to Colletotrichum falcatum reveal activation of probable effector triggered immunity (ETI) in defence responses.

KEY MESSAGE: The differential compatibility responses of sugarcane to Colletotrichum falcatum pathotypes depend on the nature of both host primary defence signalling cascades and pathogen virulence. The complex polyploidy of sugarcane genome and genetic variations in different cultivars of sugarcane remain a challenge to identify and characterise specific genes controlling the compatible and incompatible interactions between sugarcane and the red rot pathogen, Colletotrichum falcatum. To avoid host background variation in the interaction study, suppression subtractive hybridization (SSH)-based next-generation sequencing (NGS) technology was used in a sugarcane cultivar Co 7805 which is compatible with one C. falcatum pathotype but incompatible with another one. In the incompatible interaction (ICI-less virulent) 10,038 contigs were assembled from ~ 54,699,263 raw reads, while 4022 contigs were assembled from ~ 52,509,239 in the compatible interaction (CI-virulent). The transcripts homologous to CEBiP receptor and those involved in the signalling pathways of ROS, Ca2+, BR, and ABA were expressed in both interaction responses. In contrast, MAPK, ET, PI signalling pathways and JA amino conjugation related transcripts were found only in ICI. In temporal gene expression assays, 16 transcripts showed their highest induction in ICI than CI. Further, more than 17 transcripts specific to the pathogen were found only in CI, indicating that the pathogen colonizes the host tissue whereas it failed to do so in ICI. Overall, this study has identified for the first time that a probable PAMP triggered immunity (PTI) in both responses, while a more efficient effector triggered immunity (ETI) was found only in ICI. Moreover, pathogen proliferation could be predicted in CI based on transcript expression, which were homologous to Glomerella graminicola, the nearest clade to the perfect stage of C. falcatum (G. tucumanensis).

Knock-down of glucose transporter and sucrose non-fermenting gene in the hemibiotrophic fungus Colletotrichum falcatum causing sugarcane red rot.

Red rot caused by Colletotrichum falcatum, is one of the economically important disease of sugarcane and breeding for resistant varieties is considered to be the major solution to manage the disease. However, breakdown of red rot resistance become usual phenomenon due to development of newer races by culture adaptation on newly released varieties. Hence it is needed to characterize the genes responsible for pathogen virulence in order to take care of host resistance or to manage the disease by other methods. The transcript studies gave foundation to characterize the huge number of pathogenicity determinants and their role in pathogenesis. Here we studied role of two important genes viz., Glucose Transporter (GT) and Sucrose Non-Fermenting1 (SNF1) during pathogenesis of C. falcatum, which said to be involved in carbon source metabolism. Sugar metabolism has a vital role in disease progression of C. falcatum by regulating their cell growth, metabolism and development of the pathogen during various stages of infection. The present study was aimed to find out the role of GT and SNF1 genes in response to pathogenicity by RNA silencing (RNAi) approach. Knock-down of the target pathogenicity gene homologs in standard C. falcatum isolate Cf671 was carried out by amplifying sense and antisense fragments of targets individually using pSilent-1 vector. The expression cassette was cloned into the binary vector pCAMBIA1300 followed by fungal transformation through Agarobacterium mediated transformation. Resulted mutants of both the genes showed less virulence compared to wild type isolate. Simultaneously, both the mutants did not produce spores. Moreover, the molecular confirmation of the mutants displayed the expression of hygromycin gene with reduced expression of the target gene during host-pathogen interaction. Knockdown of the pathogenicity related genes (GT and SNF1) by RNAi approach corroborate the possible role of the genes in causing the disease.

Proteomic analysis of a compatible interaction between sugarcane and Sporisorium scitamineum.

Smut caused by Sporisorium scitamineum is one of the important diseases of sugarcane with global significance. Despite the intriguing nature of sugarcane, S. scitamineum interaction, several pertinent aspects remain unexplored. This study investigates the proteome level alterations occurring in the meristem of a S. scitamineum infected susceptible sugarcane cultivar at whip emergence stage. Differentially abundant proteins were identified by 2DE coupled with MALDI-TOF/TOF-MS. Comprehensively, 53 sugarcane proteins identified were related to defence, stress, metabolism, protein folding, energy, and cell division; in addition, a putative effector of S. scitamineum, chorismate mutase, was identified. Transcript expression vis-à-vis the activity of phenylalanine ammonia lyase was relatively higher in the infected meristem. Abundance of seven candidate proteins in 2D gel profiles was in correlation with its corresponding transcript expression levels as validated by qRT-PCR. Furthermore, this study has opened up new perspectives on the interaction between sugarcane and S. scitamineum.

Outbreaks of unexplained neurologic illness - Muzaffarpur, India, 2013-2014.

Outbreaks of an unexplained acute neurologic illness affecting young children and associated with high case-fatality rates have been reported in the Muzaffarpur district of Bihar state in India since 1995. The outbreaks generally peak in June and decline weeks later with the onset of monsoon rains. There have been multiple epidemiologic and laboratory investigations of this syndrome, leading to a wide spectrum of proposed causes for the illness, including infectious encephalitis and exposure to pesticides. An association between illness and litchi fruit has been postulated because Muzaffarpur is a litchi fruit-producing region. To better characterize clinical and epidemiologic features of the illness that might suggest its cause and how it can be prevented, the Indian National Centre for Disease Control (NCDC) and CDC investigated outbreaks in 2013 and 2014. Clinical and laboratory findings in 2013 suggested a noninflammatory encephalopathy, possibly caused by a toxin. A common laboratory finding was low blood glucose (<70 mg/dL) on admission, a finding associated with a poorer outcome; 44% of all cases were fatal. An ongoing 2014 investigation has found no evidence of any infectious etiology and supports the possibility that exposure to a toxin might be the cause. The outbreak period coincides with the month-long litchi harvesting season in Muzaffarpur. Although a specific etiology has not yet been determined, the 2014 investigation has identified the illness as a hypoglycemic encephalopathy and confirmed the importance of ongoing laboratory evaluation of environmental toxins to identify a potential causative agent, including markers for methylenecyclopropylglycine (MCPG), a compound found in litchi seeds known to cause hypoglycemia in animal studies. Current public health recommendations are focused on reducing mortality by urging affected families to seek prompt medical care, and ensuring rapid assessment and correction of hypoglycemia in ill children.

Pioneering superior efficiency in Methylene blue and Rhodamine b dye degradation under solar light irradiation using CeO2/Co3O4/g-C3N4 ternary photocatalysts.

In this research work, we have successfully synthesized the CeO2/Co3O4/g-C3N4 ternary nanocomposite for hydrothermal method for photocatalytic applications. The synthesized nanocomposites were characterized using X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, Field emission scanning electron microscopy (FE-SEM), Transmission electron microscopy TEM, Photoluminescent spectra (PL), X-ray photoelectron spectroscopy (XPS), Brunauer- Emmett-Teller (BET) and Ultraviolet diffuse reflectance spectroscopy (UV-DRS) technique. As per the optical spectroscopic investigations CeO2/Co3O4/g-C3N4 ternary nanocomposite exhibited the high optical absorption range and its band gap is reduced from 2.95 eV to1.83 eV. The PL spectra showed the lowered emission peak intensity of ternary nanocomposite which is revealed that the better charge separation and slow recombination of electron hole pairs. The highest photocatalytic degradation efficiency of CeO2/Co3O4/g-C3N4 ternary nanocomposite showed 93 % and 86 % towards the pollutant methylene blue and Rhodamine B. Moreover, photodegradation of the pollutants followed pseudo-first order kinetics with a very high-rate constant of 0.02211 min-1 and 0.017756 min-1. Additionally, the ternary nano catalyst was delivered the remarkable stability performance even after five cycles. This research may provide a low-cost approach for synthesized visible light responsive catalysts for use in environmental remediation applications.

Traumatic mucormycosis caused by Apophysomyces species - A case series.

Background: Apophysomyces species are emerging as the second most common agent of mucormycosis in India. It is worrisome as it affects predominantly immunocompetent host unlike other Mucorales. Unfortunately, the most common presentation is necrotising fasciitis which can be overlooked as bacterial infection. Results: Seven cases of mucormycosis due to Apophysomyces species were detected in our hospital between January 2019 and September 2022. Mean age was 55 years and all were males. Six patients presented with necrotising soft tissue infection following accidental or iatrogenic trauma. In four cases, multiple fractures over the body were observed. Median days between admission to laboratory diagnosis was 9 days. All isolates were phenotypically identified as Apophysomyces variabilis. Multiple wound debridements, on an average two, was performed in all the cases and amputation performed in two patients. Three patients recovered, two could not be treated due to financial constraints and lost to followup and two patients died. Conclusion: Through this series, we anticipate to upheave awareness among the orthopedician community about this emerging infection and contemplate it in appropriate case settings. All patients with necrotising soft tissue infection following trauma and significant degree of wound contamination with soil should be suspected for traumatic mucormycosis at the time of wound assessment.

Bacillary Angiomatosis Mimicking Pyogenic Granuloma.

Bacillary angiomatosis (BA) is an angioproliferative disease caused by Bartonella species. It manifests as nodules or papules in immunocompromised patients. Oral lesions are very rare, unlike cutaneous lesions, and histopathology plays a vital role in distinguishing these lesions from other similar ones. Treatment mainly comprises macrolides erythromycin, clarithromycin, or doxycycline.

Natural colour extraction from horticultural crops, advancements, and applications-a review.

The scope for natural colours is increasing because of the awareness of allergic, toxic, and hazardous reactions associated with synthetic dyes. Natural colours are extracted from sources that are naturally available, such as flora, fauna, and minerals. Nature sourced us multiple possibilities of colours with varied shades and hues that are subtle and harmonious when combined. Reasons like the instability of natural colours during industrial processing, seasonal availability of raw materials, fading of colour over time, cost of the benefit, and struggles in attaining sustainability have reduced commercialisation success as synthetic colours. Some plants that yield natural colours are also included in crop rotation practice. Natural dye extraction is a source of employment for the countrified subdivisions of poor developing countries. Indigenous technologies on natural colour extraction are available and have been practiced over the years; due to a lack of documentation and information on colour-yielding plants or products from horticultural crops, and their extraction methods, the use of natural colours is diminishing day by day. Even in recent years, emerging techniques have been adopted in research and development, and the information has not been brought together for the use of industries and allied sectors. Several modern approaches, such as Ultrasonication, microwave, enzymatic, supercritical, pressurised liquid extraction, etc., have proven to give better results in extracting natural colours. Thereby, having instantaneous information will help to go green, be eco-friendly, and effectively utilise all the resources without compromising industrial benefits. Reviewing the availability of natural colours from horticultural crops, classifications, recent trends in their extraction process, and applications in various fields will help achieve the above.

Anti-cariogenic property of Carica papaya, Trachyspermum ammi, Caesalpinia crista linn extracts and their effect on human oral keratinocytes.

Objectives: Dental caries is a dynamic and composite process. The multifactorial etio-pathogenesis thus influences the initiation and the progression of the disease. The prime pathogenic bacterium includes Streptococcus mutans, Lactobacillus sp and Actinomyces viscosus. The purpose of this in vitro study was to analyze the antimicrobial property of the test herbal extracts and also their effects on the human oral keratinocytes. Materials and Methods: The bacterial strains S. mutans (American Type Culture Collection [ATCC]-25175); Lactobacillus species (ATCC 4356) and A. viscosus (ATCC 15987) were cultured in the specific culture media-Mitis Salivarius Bacitracin, Man Rogosa Sharpe and Enrichment media, respectively. The test extracts were exposed to the cultured plates and the mean zone of inhibition was measured. The test herbal extracts were also tested for deleterious effects on oral keratinocytes via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Independent Student's t-test and analysis of variances were performed.-25175); Lactobacillus species (ATCC 4356) and A. viscosus (ATCC 15987) were cultured in the specific culture media-Mitis Salivarius Bacitracin, Man Rogosa Sharpe and Enrichment media, respectively. The test extracts were exposed to the cultured plates and the mean zone of inhibition was measured. The test herbal extracts were also tested for deleterious effects on oral keratinocytes via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Independent Student's t-test and analysis of variances were performed. Results: The extracts of Carica papaya, Trachyspermum ammi and Caesalpinia crista linn inhibited the growth of bacteria and the antimicrobial effect was found to be statistically significant at the neat/standard concentration (100 µg/ml). The three extracts showed a cell viability range 96%-99% indicating that the test extracts did not produce or display any deleterious effects on the oral keratinocytes. Conclusions: The three test herbal extracts possess effective anti-cariogenic properties with near par with the efficacy of chlorhexidine and T. ammi proved to be the most potent. The extracts at different concentrations also proved to be safe, noncytotoxic producing a range of 96%-99% of cell viability of the oral keratinocytes.

Paradental cyst with hyaline ring granuloma masquerading as pericoronitis.

Paradental cyst is an odontogenic cyst associated with pericoronitis in partly erupted mandibular third molars. It is an inflammatory cyst common among the mandibular molars. The cyst is most commonly seen on the distal or distobuccal aspect of the third molars. The angle of tooth and food impaction has been postulated to be responsible for the development of the cyst in third molars. The source of the epithelium has been reported as reduced enamel epithelium. The paradental cyst is frequently misdiagnosed as a radicular cyst or dentigerous cyst. We report a case of paradental cyst in a patient with partially erupted mandibular third molar with food impaction and resulting hyaline ring granuloma.

Clinicopathological characteristics of oral squamous cell carcinoma arising from oral submucous fibrosis: A systematic review.

Introduction: Oral submucous fibrosis (OSMF) is considered to be a potentially malignant oral disorder with high risk of malignant transformation. Oral squamous cell carcinoma (OSCC) arising from OSMF has peculiar clinical and histopathological features. Aim: To assess the clinicopathological features of OSCC arising in the background of OSMF in an attempt to identify the patients with OSMF who are at increased risk of developing OSCC. Methodology: A systematic review was performed based on PRISMA guidelines to include articles published until May 2021 in English, relating the clinicohistopathological characteristics of OSCC arising from OSMF (OSMF-OSCC) or OSMF associated with OSCC (OSCC-OSMF). All the eligible articles were analyzed and relevant data were extracted. Results: Seventeen articles were included for systematic review after following strict inclusion and exclusion criteria. The malignant transformation rate of OSMF-OSCC ranged from 1.9 to 9.13 and the prevalence of OSCC-OSMF ranged from 2.8 to 66. The mean age of the patients ranged from 36.6 years to 47.2 years and buccal mucosa was the common site to be affected. Majority of the OSCC-OSMF was well-to-moderately differentiated and majority of them did not metastasize to the lymph nodes. Conclusion: OSCC associated with OSMF presents at a younger age with early tumor stage, better differentiation, and better prognosis when compared to OSCC not associated with OSMF. However, more multicentric prospective studies with large sample size are required to determine the true biologic behavior of OSCC arising in the background of OSMF to establish less aggressive treatment strategies considering them as a separate entity.

Efficacy of Bovine Hydroxyapatite and Collagen Along With Platelet-Rich Fibrin as a Scaffold and Human Chorion as a Membrane for Ridge Preservation: A Case-Control Study.

AIM: The present study aims to determine the efficacy of bovine hydroxyapatite and collagen (G-graft) mixed with platelet-rich fibrin (PRF) used as a scaffold and chorion membrane as a barrier in post-extraction sockets with extraction sites alone. METHODS AND MATERIAL: Thirty individuals were randomly assigned into two groups. In the control group, after debridement of the extracted tooth socket, no additional treatment was done. In the test group, after debridement of the extracted tooth socket, the sockets were filled with bovine hydroxyapatite and collagen (G-graft) mixed with PRF. They were covered by a chorion membrane, and a non-absorbable suture material was used to secure the membrane in place. Clinical parameters assessed were plaque index, gingival index, buccolingual width, buccal bone plate height, and lingual bone plate height at baseline and at six months. RESULTS: Clinically, there was a more significant reduction in the buccolingual width of the control group than the test group after six months. A statistically significant difference between the two groups for vertical ridge height at the mesial and distal socket sites was observed. No statistically significant difference in buccal and lingual bone plate height was observed between the two groups (p>0.05). CONCLUSIONS: Both groups showed a significant reduction in the Buccolingual width, but it was less in the ridge preservation group. Thus, the use of G-graft with PRF and chorion membrane was highly effective in ridge preservation. Key messages: Natural bovine bone mineral, along with PRF and chorion as a membrane, can be utilized effectively for ridge preservation in extracted tooth sockets due to periodontal disease.

An efficient optical properties of Sn doped ZnO/CdS based solar light driven nanocomposites for enhanced photocatalytic degradation applications.

Metal sulfide - semiconductor nanocomposites synthesized with well-defined tin metal, exhibited the wide bandgap, the absorptions are limited to the UV-vis region for reduction of Reactive Blue 160 (RB 160) under solar light irradiation. The prepared samples were characterized using optoelectronic techniques. Conveniently, a wider range of wavelengths and physical properties can be enabled by doping these metal oxide nanoparticles. Whereas the photoreduction of RB 160 is unambiguously associated within charge separation and transmission progression from the excited Sn doped ZnO/CdS. Furthermore, Photocatalytic degradation efficiency for the Sn doped ZnO/CdS composites still reliant on the excitation strength, indicating the several electrons and protons were precise as a result of charge separation and transmission in prepared catalyst. Sn doped ZnO/CdS composites shows 94% Photocatalytic degradation efficiency within 120 min under sunlight irradiation. This photocatalytic nanocomposites may find capable applications in solar cells to power stretchable and also in wearable electronics.

Carbohydrate active enzymes (CAZy) regulate cellulolytic and pectinolytic enzymes in Colletotrichum falcatum causing red rot in sugarcane.

Colletotrichum falcatum, an ascomycete pathogen causes red rot of sugarcane which is specialized to infect cane stalks. Cellulolytic and pectinolytic enzymes are necessary for degradation of plant cell wall which stands as barrier for successful fungal pathogenesis. In the study, we have confined to the CAZy genes that regulate cellulolytic and pectinolytic enzymes in two distinctive pathotypes of C. falcatum. Comparative transcriptome analysis revealed that a number of CAZy genes producing cellulolytic and pectinolytic enzyme were present in the virulent (Cf671) and least virulent (RoC) pathotypes. Two consecutive transcriptome analyses (in vitro) were performed using Illumina Hi Seq 2500, further analysis was done with various bioinformatic tools. In vitro expression analysis of cutinase, glycoside hydrolyase and pectin-related genes revealed number of genes that attributes virulence. Numerous pectin-related genes involved in degradation of plant cell wall, pectinase and pectin lyase are considered to be key precursor in degradation of pectin in sugarcane. These results suggest that cellulolytic enzymes, cutinase and pectin-related genes are essential for degradation of sugarcane cell wall and considered to be an important pathogenic factor in C. falcatum. This is the first detailed report on sugarcane cell wall-degrading enzymes during its interaction with C. falcatum and also this comparative transcriptome analysis provided more insights into pathogen mechanism on C. falcatum. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-022-03113-6.

Phyto-intruders in oral tissues: A polarized light microscopic study.

OBJECTIVES: Foreign body reactions are common in the oral cavity due to its proximity to the external environment. Rarely, foreign body of plant origin may be encountered in the histopathological sections making the diagnosis problematic. The aim of the present study was to analyze the histological features of various products of plant origin emphasizing on the pathogenesis of tissue reaction occurring in response to their implantation. METHODS: This observational study included various plant products or phytoproducts commonly consumed in South Indian diet such as rice, curry leaves, coriander leaves, spinach leaves, coconut, green chilli, onion, French beans, urad dal, lentil beans, mustard seed, ginger, and garlic. Formalin-fixed specimens were routinely processed and stained with hematoxylin and eosin. The slides were viewed under light microscope and polarized microscope and evaluated by two oral pathologists. RESULTS: Each of the phytoproducts had distinctive histological appearance and exhibited positive birefringence. Phytoproducts such as rice, curry leaves, French bean, onion, and green chilli resembled pathological structures such as calcifications, ghost cells, clear cells, atypical adipocytes, and fungal hyphae, respectively. CONCLUSIONS: Plant products appear as puzzling structures in histological section posing difficulties to the pathologist. Recognition of these structures as foreign body based on their histological appearance is inevitable and their identification may avoid unnecessary delay in treatment planning. The current study serves as an atlas for the histology of extraneous material study and also as a reference for the pathologists whenever mystifying structures are encountered.

Alpha-L-fucosidase levels in patients with oral submucous fibrosis and controls: A comparative study.

Background: Oral submucous fibrosis (OSMF) in recent times has been recognized as a potentially malignant disorder (PMD) with an increased risk of developing oral squamous cell carcinoma with malignant transformation rates that vary from 0.6% to 36%. Alpha-L-fucosidase (AFU) is a lysosomal enzyme that is involved in maintaining the homeostasis of fucose metabolism. In benign and malignant tumors, the cells modulate their surface by increasing fucosylation leading to uncontrolled growth. Aims and objectives: This study was designed to estimate the levels of salivary and serum AFU in patients with OSMF and healthy controls and also to evaluate the clinical utility of salivary AFU levels over serum. Materials and Methods: Saliva and blood samples were collected from twenty participants in both the groups (OSMF and healthy controls). Serum and salivary alpha-L-fucosidase levels were measured by enzyme-linked immunosorbent assay. The data were subjected to appropriate statistical analysis. Results: We found a significant increase in alpha-L-fucosidase level in OSMF compared with healthy subjects. Pearson's correlation showed salivary alpha-L-fucosidase level to have superior sensitivity in detecting OSMF compared with serum alpha-L-fucosidase. Conclusion: The outcome of this study suggests that salivary alpha-L-fucosidase can be utilized as a biomarker in early detection of oral precancer and cancer.

Rapid economical acetic acid Papanicolaou staining procedure versus conventional staining procedure in normal oral mucosa.

Background: Papanicolaou (Pap) staining technique is a conventional technique used in cytology but it is time consuming. Aim: The aim of the study was to determine the efficacy of Rapid Economical Acetic acid Papanicolaou stain (REAP) over conventional staining technique in studying normal oral mucosa. Materials and Methods: Eighty patients were selected and were grouped based on their habits. Two smears were obtained from each patient and were subjected to both the staining techniques. A total of 160 slides were studied for features such as cellular outline, nuclear outline, nuclear details, cellular differentiation, micronuclei and cellular transparency. Results: The conventional staining procedure showed 79 cases of optimal cellular staining, 78 cases of optimal nuclear staining, 35 cases of optimal nuclear details, 57 cases of optimal differentiation and 27 cases of optimal transparency and 33 cases of optimal micronuclei. REAP staining showed 75 cases of optimal cellular staining, 64 cases of optimal nuclear staining, 20 cases of optimal nuclear details, 36 cases of optimal differentiation and 28 cases of optimal transparency and 25 cases of optimal micronuclei. Conclusion: REAP staining effectively reduces the time and the cost factor, but the cytological details are well observed under conventional staining technique in normal patients.

Emergence of novel Porcine circovirus 2 genotypes in Southern India.

A total of 200 samples from Porcine circovirus 2 suspected (n = 112) and healthy (n = 88) swine populations collected from different districts of Tamil Nadu, south India were used in this study. The samples comprising of serum (n = 124), swabs from natural orifices (n = 52), and postmortem tissues (n = 24). All the samples were processed and subjected to the screening and detection of the PCV2 genome by a specific PCR assay. PCV2 genomes from positive samples were further subjected to genotyping with specifically designed primers for the full-length amplification of the ORF2 gene which codes for capsid protein (Cp) and serves as an epidemiological marker. Randomly, 13 amplified ORF2 genes were sequenced and the aligned sequences were subjected to signature motif analysis and phylogeny in MEGA X. The molecular prevalence of PCV2 infection in Tamil Nadu is 10.5% (n = 21). Signature motif and phylogenetic studies of 13 samples revealed 38.5% (n = 5) presence of each PCV2b intermediate 1(IM1) and PCV2b genotypes, followed by 15.4% (n = 2) PCV2d-2 and 7.7% (n = 1) PCV2d genotypes. The PCV2b-IM1 genotype has a 99.43% sequence homology with Vietnam isolate (JX506730). PCV2b genotypes showed 99.72% sequence identity with Chinese isolate (KX068219). PCV2d-2 genotypes reported in this study have 100% sequence identity with Taiwan isolate (MF169721). PCV2d genotype showed 97.87% sequence identity with Thailand isolate (MF314293). Amino acid analysis of all the 13 full-length ORF2 gene sequences revealed specific mutations in the immune reactive domains of A, B, C, and D. Capsid protein of three PCV2b and five PCV2b IM1 isolates had extra amino acid residue lysine (K) at 234 position of ORF2 similar to PCV2d. For the first time in South India, PCV2b IM1 and PCV2d-2 genotypes are reported. This study evidences the genetic shifts of PCV2 isolates in India and it is analogous to that of global genotypic shift.