Role of polymerase chain reaction–based viral detection in pterygia

Purpose: Pterygium is a fibrovascular disease that originates in the conjunctiva and commonly spreads to the corneal surface, thereby posing a threat to eyesight. Despite intensive research, the pathophysiology of this disease remains unclear. Recent research suggests that oncogenic viruses, such as human papillomavirus (HPV), cytomegalovirus, and Epstein–Barr virus (EBV), may play a role in pterygia development. Although there are questions concerning the function of oncogenic viruses in pterygium pathogenesis, existing research shows a lack of consensus on the subject, demonstrating the heterogeneity of pterygium pathophysiology. Therefore, we aimed to simultaneously detect the three common viral pathogens that have been reported in pterygium tissue obtained after excision. Methods: Thirty?five tissue specimens of pterygium from patients undergoing pterygium surgery (as cases) were analyzed for evidence of viral infection with multiplex polymerase chain reaction (PCR), and virus?specific real?time quantitative PCR was used for the samples that were detected positive by multiplex PCR. Results: Of the 35 patients, one sample was positive for EBV and two samples were positive for HPV. Further PCR?based DNA sequencing of the HPV PCR?positive product showed identity with HPV?16. Real?time quantitative PCR on samples that showed EBV or HPV positivity did not yield any detectable copy number. Conclusion: Our study results confirmed that PCR positivity could be due to transient flora, but it was not quantitatively significant to conclude as the causative factor of pterygium pathogenesis. However, additional studies with larger sample populations are warranted to fully determine the role of the virus in pterygium.

Multisystem inflammatory syndrome in neonate with skin lesions—A rare case report

Multisystem inflammatory syndrome in children is a serious condition that occurs due to postinfectious immune-mediated hyperinflammatory reaction seen in children, which develops after 4–6 weeks of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection; it is rare in neonates. We present here a case of a term newborn with fever, respiratory distress, and necrotic skin lesion that gradually progressed to multisystem dysfunction. Reverse transcription polymerase chain reaction for SARS-CoV-2 was negative for both mother and the baby. SARS-CoV-2 IgG titer was negative in the mother but was found to be positive in the newborn. The diagnosis of multisystem inflammatory syndrome in neonate was made and was successfully treated with intravenous immunoglobulin and corticosteroids.

Is mean platelet volume the earliest diagnostic marker of neonatal sepsis: a prospective case control study

Background: Neonatal sepsis is the third leading cause of neonatal mortality after prematurity and intrapartum related complications worldwide. The literature regarding the use of Mean platelet volume as a diagnostic tool in neonatal sepsis is scanty.Methods: Prospective case control study in a district hospital. Neonates > 30 weeks gestation admitted to NICU during the study period were included. Neonates who did not have any risk factors for sepsis and did not have a positive blood culture or elevated CRP were categorized as group 1. Neonates who were admitted with risk factors of sepsis but did not have a positive CRP or blood culture positivity were categorized as group 2. Neonates who were admitted as probable sepsis and subsequently developed blood culture or CRP positivity were categorized as group 3.Septic workup was done for all the subjects at admission and at 72 hours after admission. Newborn with congenital anomalies and who were already on antibiotics prior to admission were excluded from the study. Statistical analysis was done using Statistical Package of Social Sciences (SPSS) version 20.0.Results: Total 240 Neonates were included in the study. Elevation of MPV in neonates with sepsis was seen as early as the first sample whereas CRP elevation was seen only on Day 3. Cut off value for Mean Platelet Volume (MPV) was found to be 10.15fl with sensitivity of 84% and specificity of 74%.Conclusions: MPV can be used as a earliest diagnostic marker for prediction of neonatal sepsis and mortality. It can facilitate early initiation of treatment without any additional exposure.

Effect of Maltodextrin Concentration and Inlet Air Temperature on Properties of Spray Dried Powder from Reverse Osmosis Concentrated Sweet Orange Juice

Abstract Sweet orange juice is an important part of diet since it is nutritious beverage offering good taste and play significant part in a healthy diet. High hygroscopicity, thermo-plasticity and presence of low molecular weight components in sweet orange juice offer low glass transition temperature (Tg), likely to form soft particle with sticky surface leading to sticky powder during drying. Maltodextrins are amorphous drying aids that tend to inhibit sugar crystallization and form a high Tg product after drying. In this study, the effect of the different spray drying parameters on the quality of powder derived from control and concentrated juice at three inlet air temperatures 120, 130 and 140 °C and at three levels of juice total soluble solids (TSS): maltodextrin levels at 1:0.5; 1:1 and 1:1.5 were studied. The impact of inlet air temperature and maltodextrin concentration has significantly affected various properties of sweet orange powder. For control juice, process yields increased with increase in inlet air temperature and maltodextrin concentration. However, for reverse osmosis (RO) concentrate, process yield increased with increase in maltodextrin concentration and decreased with increase in inlet air temperature. For control juice, process yields obtained were in the range of 12.59-41.16% and in case of concentrated juice, the process yield obtained was in the range of 21.35-56.95% at different combinations of inlet air temperature and maltodextrin concentrations. Spray-dried powder was considered as "possible" and "fair" in terms of flowability and cohesiveness. Vitamin C retention was high at lower inlet air temperature with lower concentration of maltodextrin.

Subconjunctival bevacizumab and argon laser photocoagulation for preexisting neovascularization following deep lamellar anterior keratoplasty

We report a rare case of deep anterior lamellar keratoplasty (DALK) neovascularization managed with combination of subconjunctival bevacizumab and argon laser photocoagulation. A 24 year old male underwent Deep anterior lamellar keratoplasty for corneal stromal opacity following presumed viral keratitis. Deep corneal neovascularization was observed postoperatively which was successfully managed using a combination of subconjunctival bevacizumab and argon laser photocoagulation within one week of DALK. The neovascularization resolved by 3 months and at 2 years follow up, patient maintained good visual acuity of 6/12 Snellen's without recurrence of vascularization. A combination of bevacizumab and argon laser may be an effective approach to manage neovascularisation in the immediate postoperative phase (Post DALK) and improve graft survival.

Unusual ulcerative keratitis caused by Prototheca wickerhamii in a diabetic patient

The purpose of the study was to report a case of ulcerative keratitis caused by an unusual algae Prototheca wickerhamii in a diabetic patient. This study design was a case report. A 46-year-old male, who was a known diabetic for 3 years, had an injury to the left cornea with the sparks of fire from wielding at work that developed into an ulcerative keratitis over a period of next 3 months as the patient was not on any medication. Corneal scraping culture report and Vitek 2 system investigation result confirmed it to be a P. wickerhamii infection. The patient was started on intensive topical 1% voriconazole and 5% natamycin for 1 month and with no improvement subsequently underwent penetrating keratoplasty. No recurrence of infection postoperatively was noted. This opportunistic algae rarely known to cause human eye infections is so far reported in either patients with severe systemic immunosuppression causing posterior segment eye involvement or as postcorneal surgery infections. We report an ulcerative keratitis by P. wickerhamii in a diabetic patient post corneal trauma with no prior ocular surgery.

Nested reverse transcriptase-polymerase chain reactions targeting the messenger RNA of icl[2], hspx, and rRNAP1 genes to detect viable Mycobacterium tuberculosis directly from clinical specimens

There is an urgent need for a rapid and reliable test to detect actively multiplying Mycobacterium tuberculosis directly from clinical specimens for an early initiation of the appropriate antituberculous treatment. This study was aimed at the optimization and application of nested reverse transcriptase-PCR [nRT-PCR] targeting the messenger RNA of the icl[2], hspx, and rRNAP1 genes directly from sputum specimens, and their evaluation against the culture by the BACTEC MicroMGIT mycobacterial culture system. 203 Sputum samples from clinically suspected tuberculosis patients and 30 control specimens [clinically proven viral or bacterial infections other than tuberculosis] were included in this study. The mycobacterial culture was performed by the BACTEC MicroMGIT system following the manufacturer's instructions. The primers for nRT-PCRs targeting icl[2], hspx, and rRNAP1 genes were indigenously designed using the Primer-BLAST software, and optimized for sensitivity and specificity. The icl[2], hspx, and rRNAP1 genes were able to pick up 63.9%, 67.2%, and 58.75%, respectively, of culture-negative sputum specimens collected from clinically suspected tuberculosis patients. However, three [1.4%] were negative for nRT-PCR, but M. tuberculosis culture positive. All the 30 controls were negative for culture by the BACTEC MicroMGIT method and all three nRT-PCR. The novel nRT-PCRs targeting icl[2], hspx, and rRNAP1 genes developed in this study are rapid and reliable diagnostic tools to detect viable M. tuberculosis directly from sputum specimens. However, further study by including a larger number of sputum specimens needs to be carried out to ascertain the diagnostic utility of the novel nRT-PCRs optimized in the study

Genetic analysis of HA gene of pandemic H1N1 2009 influenza viruses circulating in India.

The H1N1 2009 influenza pandemic took the health care workers by surprise in spite of warning about influenza pandemic. Influenza A virus has the ability to overcome immunity from previous infections through the acquisition of genetic changes by shift or drift. Thus, understanding the evolution of the viruses in human is important for the surveillance and the selection of vaccine strains. A total of 23 pandemic A/H1N1 2009 viral HA gene sequences were downloaded from NCBI submitted during March and May 2010 by NIV and were analysed. Along with that the vaccine strain A/California/07/2009 was also downloaded from NCBI. All the sequences were used to analyse the evolution of the haemagglutinin (HA) by phylogenetic analysis. The HA gene could be divided into four groups with shift from 1 to lV revealing that the HA genes of the influenza A viruses evolved in a sequential way, in comparison to vaccine strain A/California/07/2009. Amino acid sequence analysis of the HA genes of the A/H1N1 2009 isolates, revealed mutations at positions 100, 220 and additional mutations in different positions 114, 171, 179, 190, 208, 219, 222, 239, 240, 247, 251, 260 and 285 .The mutations identified showed the adaptation of the new virus to the host that could lead to genetic changes inherent to the virus resulting in a reassortant which could be catastrophic, hence continuous monitoring of strains is mandatory.

Comparison of manual and physiologically molded denture bases in complete denture wearers.

Background and Objectives: In complete denture wearers, the accumulation of food in the buccal vestibule is one of the factors causing discomfort to the patient. This study was conducted to evaluate the effect of mandibular denture buccal flange on food retention and to correlate between varying angulations of buccal flange of mandibular denture on food retention. Materials and Methods: Complete dentures were fabricated and the mandibular complete denture was duplicated. The existing (unduplicated) manually molded and physiologically molded dentures were tested for food retention. Twenty test particles having a diameter of 2±0.2 mm were placed on the middle part of the buccal flange and the subjects were instructed to expel the test particles by performing only functional movements of the buccal musculature. The numbers of residual test particles in the buccal vestibule were counted after 30s. The angulations of the denture buccal flange (duplicated) were measured with an optical bevel protractor and trimmed to 50° and the food retention test was performed. Similar tests were performed at 60°, 70° and 80° angulations. The data were evaluated using the Pearson correlation coefficient, One-way ANOVA test and Wilcoxon signed rank test. Results: The difference between the residual scores of manually molded and physiologically molded denture base was not significant. The difference between the residual scores at different angulations of the buccal flange of the denture was significant. Conclusion: Only the slope of the flange significantly affects the ability of subjects to expel the test particles.

Evaluation, grafting success and field establishment of cashew rootstock as influenced by VAM fungi.

Seven isolates of vesicular arbuscular mycorrhizal (VAM) fungi were isolated from cashew rhizosphere soil of different cashew growing regions of South India. These seven isolates along with two more VAM fungi namely Acaulospora laevis and Glomus mosseae, which were found to be better symbionts for cashew during our earlier study were used to study their effectiveness on the growth and nutrition of cashew rootstock Ullal-1. Four promising VAM fungi were selected based on this study. Rootstocks inoculated with these four fungi were evaluated for their vigour through grafting success, using Ullal-3 cashew variety as scion. Grafting success was more in rootstocks inoculated with A. laevis and one of local isolates Glomus etunicatum. Grafts with rootstock treated with G. etunicatum and A. laevis survived and performed better when planted in the field compared to the uninoculated and other VAM fungal treatments.

Characterization of leprosy based on the nasal lipid profile.

While extracting the M. leprae from the nasal flushings of leprosy patients it was found that these organisms were trapped in the waxy layer, between the aqueous and the chloroform layers. Thin layer chromotography (TLC) analysis of this layer, using chloroform-methanol-water system, revealed different spots when sprayed with acid alcohol and heated at 160 degrees C. The TLC profile of lipids of lepromatous and borderline (MB according to the WHO terminology) leprosy patients was distinctly different from that of tuberculoid leprosy patients and normal human volunteers. A simple, economical and fast procedure to characterize patients belonging to different spectra has been developed.

Bone-transplants (heterogenous)

It is not intended here to describe the fate and ultimate absorption of the bone peg in detail, although it has been observed that the whole process of final incorporation of the peg takes place without much of discomfort into the recepient bone. No foreign protein reaction was ever observed, and also the peg was never seen to extrude. The only precaution required to be observed was that the peg should not exceed 3 mm in diameter and 4 to 5 cm in length. Any excess in the size of the bone peg other than the stated measurements, leads in my opinion, to a foreign body reaction and it is found that there is a surrounding inflammatory reaction, which is not due to a septic process. Hence the limitations of use is evident. As cited in the cases above, one thing stands out in common, and that is, the peg must be slender enough to be incorporated into the medullary cavity of a metacarpal or that of a radius, ulna or a drill hole of 2.5 mm in the case of medial malleolus of medial epicondyle of the humerus. Therefore, it is clear, that no extensive beef-bone plate or other stout pegs can be used with success. It is pointed out, that in these selected variety of fractures, use of a slender beef-bone peg is met with good results and good union. The following X-Ray pictures denote where the beef-bone pegs were used with satisfactory results.