A Study of Cytomorphological Spectrum of Head and Neck Lesions in Pediatric Age Group.

Introduction: Fine Needle Aspiration Cytology (FNAC) is a well-established early diagnostic technique for evaluating mass lesions in adult patients. Now, FNAC in children is gaining acceptance and is used as a first-line investigation in diagnosis of pediatric lesions. Aims: To analyze the cytomorphologic spectrum of head and neck lesions in pediatric age group with histopathological correlation wherever possible and to study the utility of FNAC in pediatric head and neck lesions. Materials and Methods: A prospective study was carried out on all FNACs of head and neck lesions in pediatric age group (0-18 years), detected clinically or under radiological guidance for a period of 3 years from August 2018 to July 2021. Results: The study included 238 cases. Most of the cases were seen in the age group of 13-18 years and with male to female ratio of 1.35:1. Most common site of FNAC was lymph nodes (70.2%) and the commonest lesion encountered was reactive lymphadenitis (50.8%). Second most common site was thyroid (15.9%). Soft tissue/bone, salivary gland, miscellaneous/skin lesions were also encountered. Among the 43 neoplastic lesions, benign (31 cases) were more common than the malignant (12 cases). The malignant cases included non-Hodgkins lymphoma, Hodgkins lymphoma, metastasis to lymph node, low-grade sarcoma, papillary thyroid carcinoma, and Langerhans cell histiocytosis of bone. Histopathological correlation was done in 32 cases (13.4%). Statistical analysis showed a sensitivity of 85.29% and specificity of 97.74%. Overall diagnostic accuracy was 96.3%. Conclusion: This study highlighted various cytomorphological patterns in head and neck lesions with high diagnostic accuracy in children. FNAC helps in proper planning of treatment modalities in head and neck masses in pediatric age group.

Appendicitis Inflammatory Response Score in Comparison to Alvarado Score in Acute Appendicitis.

Appendicitis is a common differential diagnosis of right lower quadrant pain. Clinical evaluation alone results in high negative appendicectomy rates. Alvarado scoring is the most commonly used clinical prediction rule. The study aimed to compare the recently developed appendicitis inflammatory response (AIR) score with the Alvarado score. This cross-sectional observational study included patients who underwent appendicectomy for clinical suspicion of appendicitis. The clinical and laboratory parameters required for obtaining Alvarado score and AIRS were gathered. Area under ROC curve was calculated for both Alvarado score and AIRS. The study included 130 patients (77 males and 53 females). The negative appendicectomy rate was 10.7%. The perforation rate was 10.3%. The area under ROC for Alvarado score was 0.821 and for AIR score was 0.901. The Alvarado score had a sensitivity of 72% and a specificity of 79% at score ≥6. The appendicitis inflammatory response score had a sensitivity of 98% for scores ≥5 and a specificity of 97% for score ≥6. The C-reactive protein (CRP) value was the best performing individual parameter with an area under ROC of 0.789, followed by WBC count with an area under ROC of 0.762. Appendicitis inflammatory response score is a recently developed score that outperforms the Alvarado score. AIR score has a higher specificity. The sound construction, gradation of parameters, the inclusion of CRP, and avoidance of subjective parameters make the AIR score an attractive clinical prediction rule which can decrease the rate of negative appendicectomy.

Chronic blepharitis like picture in patients with Discoid lupus erythematosis - Case series.

BACKGROUND: Blepharitis is a very common condition encountered in ophthalmology outpatient care department. Dermatological diseases like seborrheic keratopathy and rosacea have been frequently discussed as being associated with blepharitis. However Discoid lupus erythematosis [DLE], an autoimmune condition has only rarely been reported to involve the eyelids mimicking blepharitis. DLE affecting the eyelids can produce significant morbidity with lid deformities, trichiasis and symblepharon if left untreated. OBJECTIVE: To report three consecutive cases of DLE with eyelid lesions mimicking blepharitis. They presented to the department of ophthalmology at our institute from April 2014 to March 2016. The first case presented with involvement of lower eyelid in one eye. She was having multiple skin lesions which on biopsy confirmed the diagnosis of DLE. The second case was a diagnosed case of DLE who received treatment 4 years back and came with relapse of the disease affecting the eyelid. The third case was a recently biopsy confirmed case of DLE with multiple skin lesions along with bilateral eyelid involvement. Two of them had madarosis and one case had destruction of the outer lid margin at presentation. All three cases responded well to treatment with Hydrxychloroquin. CONCLUSION: The discoid lesions of DLE affecting the eyelids can mimic the appearance of chronic blepharitis. Ophthalmologists should be aware that DLE is a possibility while dealing with an atypical case of chronic blepharitis. Early diagnosis and treatment can prevent deformities of eyelid.

Purification and antibiofilm activity of AHL-lactonase from endophytic Enterobacter aerogenes VT66.

The opportunistic pathogen Pseudomonas aeruginosa uses biofilm lifestyle to resist antibiotic treatment. In our study, endophytic bacterium Enterobacter aerogenes VT66 quenched the N-acyl homoserine lactone (AHL) molecules produced by P. aeruginosa PAO1. The quorum quenching activity was attributed to the presence of AHL-lactonase. The AHL-lactonase was purified using column chromatography and purified AHL-lactonase was applied for the control of biofilm formation in P. aeruginosa PAO1. The results showed that purified AHL-lactonase obtained with a molecular weight about 30kDa was able to inhibit more than 70% of biofilm in P. aeruginosa PAO1 (P<0.001). Antibiofilm activity of AHL-lactonase was correlated well with results from staining technique used to determine inhibition of biomass and viable cell activity. Therefore, results unambiguously confirm that the AHL-lactonase from E. aerogenes VT66 could be used as antibiofilm therapeutics in P. aeruginosa associated biomedical applications.

In vitro biological activity of aromadendrin-4'-methyl ether isolated from root extract of Ventilago madraspatana Gaertn with relevance to anticandidal activity.

In this study, antimicrobial and antioxidant activities of the hexane extract of the root of Ventilago madraspatana were evaluated. Based on the significant bioactivity of crude hexane extract, an active compound was purified from the root extract. The active compound was further purified and identified as aromodendrin-4'-methyl ether by the (1)H NMR spectrum. The isolated compound significantly inhibited Staphylococcus epidermidis with the lowest MIC and MBC at 78 µg/mL (P < 0.05). The compound also exhibited significant anticandidal activity with MIC and MBC values of 312 and 625 µg/mL, respectively. The radical scavenging activity of aromodendrin-4'-methyl ether was evident by its lower IC50 values of 60 µg/mL for DPPH scavenging and 3.2 µg/mL for ABTS scavenging. The compound also exhibited ferrous ion chelation and H2O2 scavenging activities. The study is an attempt to increase the industrial utility of V. madrasapatana.

Use of aiiA gene amplification for AHL-lactonase production from endophytic bacterium Enterobacter species.

AHL-lactonase has gained renewed interest due to biotechnological applications such as antiquorum sensing, antibiofilm strategies, biofouling, etc. In our study, the production of AHL-lactonase from endophytic bacteria Enterobacter aerogenes VT66 was optimized by response surface methodology (RSM) using central composite design (CCD) for four different cultural conditions. The relative activity of AHL-lactonase was correlated with amplification of aiiA homologous gene amplification with respect to cultural conditions. Statistical analysis by ANOVA of the quadratic regression model showed that the RSM model constructed is highly significant, as indicated by F-test with a low probability value (p(model) < 0.0001) and high regression coefficient (0.9997) as well as lower coefficient of variation (1.86%) indicate that suitability of variable parameters. The quadratic regression model of AHL-lactonase production in terms of relative activity was built and the optimal cultural conditions for maximum enzyme production were determined as 32.5 °C temperature, pH 7.0, 350 µM of substrate concentration and 33 h of incubation time. The enhanced AHL-lactonase yielded 1.33 fold increases in relative activity and it positively correlated with the amplification of aiiA gene.

Molecular identification of aiiA homologous gene from endophytic Enterobacter species and in silico analysis of putative tertiary structure of AHL-lactonase.

The aiiA homologous gene known to encode AHL- lactonase enzyme which hydrolyze the N-acylhomoserine lactone (AHL) quorum sensing signaling molecules produced by Gram negative bacteria. In this study, the degradation of AHL molecules was determined by cell-free lysate of endophytic Enterobacter species. The percentage of quorum quenching was confirmed and quantified by HPLC method (p<0.0001). Amplification and sequence BLAST analysis showed the presence of aiiA homologous gene in endophytic Enterobacter asburiae VT65, Enterobacter aerogenes VT66 and Enterobacter ludwigii VT70 strains. Sequence alignment analysis revealed the presence of two zinc binding sites, "HXHXDH" motif as well as tyrosine residue at the position 194. Based on known template available at Swiss-Model, putative tertiary structure of AHL-lactonase was constructed. The result showed that novel endophytic strains of Enterobacter genera encode the novel aiiA homologous gene and its structural importance for future study.

Quorum quenching activity in cell-free lysate of endophytic bacteria isolated from Pterocarpus santalinus Linn., and its effect on quorum sensing regulated biofilm in Pseudomonas aeruginosa PAO1.

Quorum sensing mechanism allows the microorganisms to resist the antibiotic treatment by forming biofilms. Quorum quenching is one of the mechanisms to control the development of drug resistance in microbes. Endophyte bacteria are beneficial to plant growth as they support the immune system against the pathogen attack. The endophytic bacteria present in Pterocarpus santalinus were screened for the presence of N-acyl homoserine lactones (AHLs) degrading bacteria using biosensor strains and further confirmed by quantifying the violacein production. Cell-free lysate of endophytic bacteria, Bacillus firmus PT18 and Enterobacter asburiae PT39 exhibited potent AHL degrading ability by inhibiting about 80% violacein production in biosensor strain. Furthermore, when the cell-free lysate was applied to Pseudomonas aeruginosa PAO1 and PAO1-JP2 biofilm it resulted in significant (p<0.01) inhibition of biofilm formation. The biofilm inhibition was confirmed by visualization of biofilm slides under fluorescence microscopy, which showed decrease in total biomass formation in treated slides. Isolation and amplification of the gene (aiiA) indicated that the presence of AHL lactonase in cell-free lysate and sequence alignment indicated that AiiA contains a "HXHXDH" zinc-binding motif that is being conserved in several groups of metallohydrolases. Therefore, the study shows the potential of AHLs degradation by AHL lactonase present in cell-free lysate of isolated endophytic bacteria and inhibition of quorum sensing regulated biofilm formation in P. aeruginosa PAO1.

Comparison of endothelial cell loss and surgically induced astigmatism following conventional extracapsular cataract surgery, manual small-incision surgery and phacoemulsification.

PURPOSE: To compare the surgically induced astigmatism (SIA) and endothelial cell loss following conventional extracapsular cataract surgery (ECCE), manual small-incision cataract surgery (Blumenthal technique)(SICS) and phacoemulsification (PE) with non-foldable intraocular lens implantation. METHODS: 186 cataractous eyes with nuclear sclerosis grade 3 or less were randomized to undergo ECCE, SICS or PE with intraocular lens (non-foldable) implantation after a detailed pre-operative assessment. Keratometry and specular microscopy were performed pre-operatively and 6 weeks postoperatively. Surgically induced astigmatism was calculated using the rectangular coordinate method (Holladay et al.). RESULTS: Mean endothelial cell loss was similar for all three groups (p = 0.855); ECCE induced a loss of 4.72% (SD: 13.07); SICS 4.21% (SD: 10.29) and PE 5.41% (SD: 10.99). Mean SIA was 1.77D (1.61D) for the ECCE group, 1.17D (0.95D) for the SICS group and 0.77D (0.65D) for the PE group (p = 0.001). The magnitude of the difference between the SICS and the PE group was 0.4D. CONCLUSION: PE induced less astigmatism than SICS and ECCE in this study but the magnitude of the difference between SICS and PE was small. There was no significant difference in endothelial cell loss between the three groups.