Effects of PNA Sequence and Target Site Selection on Function of a 4.5S Non-Coding RNA.

Peptide nucleic acid (PNA) based antisense strategy is a promising therapeutic approach to specifically inhibit target gene expression. However, unlike protein coding genes, identification of an ideal PNA binding site for non-coding RNA is not straightforward. Here, we compare the inhibitory activities of PNA molecules that bind a non-coding 4.5S RNA called SRP RNA, a key component of the bacterial signal recognition particle (SRP). A 9-mer PNA (PNA9) complementary to the tetraloop region of the RNA was more potent in inhibiting its interaction with the SRP protein, compared to an 8-mer PNA (PNA8) targeting a stem-loop. PNA9, which contained a homo-pyrimidine sequence could form a triplex with the complementary stretch of RNA in vitro as confirmed using a fluorescent derivative of PNA9 (F-PNA13). The RNA-PNA complex formation resulted in inhibition of SRP function with PNA9 and F-PNA13, but not PNA8 highlighting the importance of target site selection. Surprisingly, F-PNA13 which was more potent in inhibiting SRP function in vitro, showed weaker antibacterial activity compared to PNA9 likely due to poor cell penetration of the longer PNA. Our results underscore the importance of suitable target site selection and optimum PNA length to develop better antisense molecules against non-coding RNA.

Giant Pleomorphic Adenoma of Infratemporal Fossa: A Rare Case Report.

Of all the head and neck tumors, salivary gland tumors account to 3%. Pleomorphic adenomas are one of the most common benign tumors arising from major salivary glands, although it could also develop from minor salivary glands situated at accessory sites like nasal cavity, pharynx, parapharyngeal space, lacrimal glands etc. Tumors of infratemporal fossa are quite unusual, mainly because of its hidden location in retromaxillary region. We report an unusual case of 65 years old male presenting with complaint of progressive left cheek swelling for 4 years. FNAC revealed pleomorphic adenoma of minor salivary gland tumor. Intraoperatively a giant lobulated tumor was seen occupying almost whole space of infratemporal fossa, which was removed in-toto via open approach. Patient was kept on regular follow up with no evidence of recurrence reported till date.

Outcomes of Palisade Cartilage Tympanoplasty in Chronic Otitis Media: Our Experience at a Tertiary Care Centre.

To evaluate the anatomical and functional success rates of palisade cartilage tympanoplasty in chronic otitis media. Thirty patients with chronic otitis media with subtotal perforation underwent palisade cartilage tympanoplasty and were assessed prospectively. The outcomes evaluated were graft uptake and hearing gain. Overall graft uptake was 90% with failure in 3 cases. The preoperative mean air conduction threshold was 22.087 ± 6.120 dB which was improved to 13.387 ± 5.253 dB postoperatively at 12 weeks which was statistically significant. The mean postoperative ABG closure was 8.700 dB with a p value of 0.001 which was statistically significant. Palisade cartilage tympanoplasty demonstrates that subtotal perforations, which are at high risk for graft failure, can be treated efficiently and a durable and resistant reconstruction of the tympanic membrane with reasonable hearing can be achieved.

Evaluation of Pre and Post-operative Pulmonary Artery Pressures in Children with Adenotonsillar Hypertrophy and Correlation with Chronicity: A Prospective Study on 50 Children.

Children with grade III or IV of adenotonsillar hypertrophy especially of a longer duration, may show cardiovascular changes due to mouth breathing. These changes can be detected on doppler echocardiocardiography which can be used as a screening tool. Fifty paediatric patients of grade 3 and 4 adenotonsillar hypertrophy underwent adenotonsillar resection after a pre-operative work up which included doppler echocardiography to measure pulmonary artery pressures. A post-operative doppler echocardiography was done after three months. 11 children (22%) showed deranged pulmonary artery pressure values with increased mean pulmonary artery pressure (MPAP) and pulmonary artery systolic pressure (PSAP) on preoperative doppler echocardiography, and 2 out of these 11 children had Pulmonary Artery Hypertension. A significant (p < 0.001) improvement was noted in pulmonary artery pressure values (MPAP and PSAP returned to normal) after adenotonsillectomy in 3-month postoperative echocardiography. Also, a statistically significant correlation was obtained between chronicity of symptoms and raised pulmonary artery pressures. Our study concluded that these children with chronic untreated upper airway obstructive symptoms get predisposed to potential cardiac complications. And these grave consequences can be easily prevented by a timely intervention.

Temporal Bone Osteomyelitis Masquerading as Malignancy: A Diagnostic Challenge.

Xanthogranulomatous osteomyelitis is a rare chronic inflammatory disorder. Until now, it has only been reported in long bones. To the best of our knowledge, it has never been reported in temporal bone. We present the case of this rare disease in a 64-year-old male involving the temporal bone, presenting with ear pain, discharge, decreased hearing, and granulation tissue in the external auditory canal, mimicking malignancy clinically and radiologically. The patient was unresponsive to medical management and was taken up for surgical debridement, followed by treatment with systemic and topical antibiotics, with a successful outcome. As this disease has not been reported in the literature yet in the temporal bone and mimics malignancy, it must be differentiated on histopathology to establish a definite diagnosis and provide appropriate management. A long-term follow-up is also necessary to recognize the clinical behavior of this disease, as no treatment protocol has been established yet.

Surgical Outcomes of Type 1 Tympanoplasty in Chronic Otitis Media in Paediatric Patients: A Prospective Study.

Objectives: To observe the surgical outcome of type 1 tympanoplasty in paediatric patients in the form of graft uptake and audiological improvement. Materials and methods: A prospective study was done in which 40 paediatric patients in age group 6-16 years, diagnosed of chronic otitis media, were taken up for tympanoplasty under general anaesthesia. Patients were followed in post operative period at 4, 6 and 12 weeks for assessment of graft status and audiological evaluation. Results: In this study, we found surgical success rate in the form of graft uptake in 80% of the cases and audiological improvement in 75% of the cases. Conclusion: Tympanoplasty is safe and effective in children with adequate graft uptake and audiological outcome. Supplementary Information: The online version contains supplementary material available at 10.1007/s12070-023-03965-1.

A Rare Presentation of Thyroid Mass as Tubercular Abscess: A Case Report.

Isolated thyroid gland tuberculosis is a rare disease, while even rarer is its presentation as an acute abscess. Here we present the case of a 65-year-old female who presented with a neck swelling which progressed to an abscess with multiple discharging sinuses which was proven to be tubercular abscess on histopathology.

Effect of Chronic Suppurative Otitis Media on Gustatory Function of chorda Tympani Nerve : A Prospective Study.

Introduction the ability to taste is among the life's finest pleasures and perception of taste can be altered by various disorders or therapeutic regimens as colds or dental procedures. Head trauma patients, patients on certain drugs such as phenylbutanone, carbamazepine and baclofen and patients with inflammatory diseases of the middle ear frequently complain about alterations in their ability to taste. So, assessment of gustatory sensitivity is the prerequisite for the correct diagnosis of taste dysfunction which uses taste strips impregnated with taste stimulants. Material and method in this prospective study, 40 patients were enrolled after assessment of history, general and local examination of ear and finally assessment of gustatory function was done according to Muller technique using taste strips. Results in present study out of 40 patients, 16 (40%) patients had COM squamous, 24 patients (60%) had COM mucosal. In the ipsilateral diseased ear (both COM squamous & mucosal) the mean taste score for various taste sensation was 11.65 ± 2.59. In the contralateral normal ear, the mean taste score was 15.42 ± 0.78. The difference between the two was found to be statistically significant (p < 0.0001). Conclusion a significant difference was obtained in taste scores between diseased (ipsilateral) and normal (contralateral) ears both overall and also when subtypes were compared. Thus, the present prospective study indicates that COM squamous as well as mucosal can induce taste dysfunction. Supplementary Information: The online version contains supplementary material available at 10.1007/s12070-023-03703-7.

Post-Covid Isolated Mandibular Mucormycosis in a Patient with Chronic Kidney Disease: A Case Report.

Mucormycosis is an opportunistic fungal infection which was earlier seen only in immunocompromised patients. With the recent covid pandemic, there had been a drastic surge in cases of mucormycosis worldwide and especially in India. Here, we present an unusual case of biopsy proven invasive mandibular mucormycosis in a patient with chronic kidney disease and a history of COVID-19 infection. The patient was given low-dose amphotericin B and underwent surgical debridement with a successful outcome.

Marked elevations in lung and plasma ceramide in COVID-19 linked to microvascular injury.

The pathogenesis of the marked pulmonary microvasculature injury, a distinguishing feature of COVID-19 acute respiratory distress syndrome (COVID-ARDS), remains unclear. Implicated in the pathophysiology of diverse diseases characterized by endothelial damage, including ARDS and ischemic cardiovascular disease, ceramide and in particular palmitoyl ceramide (C16:0-ceramide) may be involved in the microvascular injury in COVID-19. Using deidentified plasma and lung samples from COVID-19 patients, ceramide profiling by mass spectrometry was performed. Compared with healthy individuals, a specific 3-fold C16:0-ceramide elevation in COVID-19 patient plasma was identified. Compared with age-matched controls, autopsied lungs of individuals succumbing to COVID-ARDS displayed a massive 9-fold C16:0-ceramide elevation and exhibited a previously unrecognized microvascular ceramide-staining pattern and markedly enhanced apoptosis. In COVID-19 plasma and lungs, the C16-ceramide/C24-ceramide ratios were increased and reversed, respectively, consistent with increased risk of vascular injury. Indeed, exposure of primary human lung microvascular endothelial cell monolayers to C16:0-ceramide-rich plasma lipid extracts from COVID-19, but not healthy, individuals led to a significant decrease in endothelial barrier function. This effect was phenocopied by spiking healthy plasma lipid extracts with synthetic C16:0-ceramide and was inhibited by treatment with ceramide-neutralizing monoclonal antibody or single-chain variable fragment. These results indicate that C16:0-ceramide may be implicated in the vascular injury associated with COVID-19.

Tensions in Taxonomies: Current Understanding and Future Directions in the Pathobiologic Basis and Treatment of Group 1 and Group 3 Pulmonary Hypertension.

In the over 100 years since the recognition of pulmonary hypertension (PH), immense progress and significant achievements have been made with regard to understanding the pathophysiology of the disease and its treatment. These advances have been mostly in idiopathic pulmonary arterial hypertension (IPAH), which was classified as Group 1 Pulmonary Hypertension (PH) at the Second World Symposia on PH in 1998. However, the pathobiology of PH due to chronic lung disease, classified as Group 3 PH, remains poorly understood and its treatments thus remain limited. We review the history of the classification of the five groups of PH and aim to provide a state-of-the-art review of the understanding of the pathogenesis of Group 1 PH and Group 3 PH including insights gained from novel high-throughput omics technologies that have revealed heterogeneities within these categories as well as similarities between them. Leveraging the substantial gains made in understanding the genomics, epigenomics, proteomics, and metabolomics of PAH to understand the full spectrum of the complex, heterogeneous disease of PH is needed. Multimodal omics data as well as supervised and unbiased machine learning approaches after careful consideration of the powerful advantages as well as of the limitations and pitfalls of these technologies could lead to earlier diagnosis, more precise risk stratification, better predictions of disease response, new sub-phenotype groupings within types of PH, and identification of shared pathways between PAH and other types of PH that could lead to new treatment targets. © 2023 American Physiological Society. Compr Physiol 13:4295-4319, 2023.

Characterization of pulmonary vascular remodeling and MicroRNA-126-targets in COPD-pulmonary hypertension.

BACKGROUND: Despite causing increased morbidity and mortality, pulmonary hypertension (PH) in chronic obstructive pulmonary disease (COPD) patients (COPD-PH) lacks treatment, due to incomplete understanding of its pathogenesis. Hypertrophy of pulmonary arterial walls and pruning of the microvasculature with loss of capillary beds are known features of pulmonary vascular remodeling in COPD. The remodeling features of pulmonary medium- and smaller vessels in COPD-PH lungs are less well described and may be linked to maladaptation of endothelial cells to chronic cigarette smoking (CS). MicroRNA-126 (miR126), a master regulator of endothelial cell fate, has divergent functions that are vessel-size specific, supporting the survival of large vessel endothelial cells and inhibiting the proliferation of microvascular endothelial cells. Since CS decreases miR126 in microvascular lung endothelial cells, we set out to characterize the remodeling by pulmonary vascular size in COPD-PH and its relationship with miR126 in COPD and COPD-PH lungs. METHODS: Deidentified lung tissue was obtained from individuals with COPD with and without PH and from non-diseased non-smokers and smokers. Pulmonary artery remodeling was assessed by ⍺-smooth muscle actin (SMA) abundance via immunohistochemistry and analyzed by pulmonary artery size. miR126 and miR126-target abundance were quantified by qPCR. The expression levels of ceramide, ADAM9, and endothelial cell marker CD31 were assessed by immunofluorescence. RESULTS: Pulmonary arteries from COPD and COPD-PH lungs had significantly increased SMA abundance compared to non-COPD lungs, especially in small pulmonary arteries and the lung microvasculature. This was accompanied by significantly fewer endothelial cell markers and increased pro-apoptotic ceramide abundance. miR126 expression was significantly decreased in lungs of COPD individuals. Of the targets tested (SPRED1, VEGF, LAT1, ADAM9), lung miR126 most significantly inversely correlated with ADAM9 expression. Compared to controls, ADAM9 was significantly increased in COPD and COPD-PH lungs, predominantly in small pulmonary arteries and lung microvasculature. CONCLUSION: Both COPD and COPD-PH lungs exhibited significant remodeling of the pulmonary vascular bed of small and microvascular size, suggesting these changes may occur before or independent of the clinical development of PH. Decreased miR126 expression with reciprocal increase in ADAM9 may regulate endothelial cell survival and vascular remodeling in small pulmonary arteries and lung microvasculature in COPD and COPD-PH.

Association between Hypodontia and Angle's Malocclusions among Orthodontic Patients in Kathmandu, Nepal.

Background: Disturbances during the early tooth development stages may result in the congenital absence of teeth. The purpose of this study was to assess the relationship between hypodontia and Angle's malocclusions. Materials and Methods: The sample comprised 601 orthodontic patients' pretreatment records (242 men and 259 women), selected from the achieved orthodontic records. Developmental anomalies of teeth affecting the number were examined on dental panoramic radiographs. Based on Angle's classification, pretreatment dental casts were assessed and classified into different classes of malocclusion. The relationship between hypodontia and different classes of malocclusion was evaluated using the chi-square test. Results: The prevalence of tooth agenesis was 7.48%, that is, 45 out of 601 samples. There were a total of 72 (0.42%) missing teeth, excluding the third molars. The most frequent missing tooth was the maxillary lateral incisor (35, 48.61%), followed by the mandibular lateral incisor (14, 19.44%), the mandibular central incisor (6, 8.33%), the mandibular second premolar (5, 6.294%), and the maxillary second premolar (4, 5.55%). Hypodontia was more common in the upper jaw. Although hypodontia was mostly seen in Class I malocclusion patients (7.87%), followed by Class II malocclusion patients (6.99%) and least in Class III malocclusion patients. However, there was no significant difference in hypodontia among different classes of malocclusions (p = 0.352). Conclusion: The most frequently missing tooth was the maxillary lateral incisor, followed by lateral and central mandibular incisors and mandibular second premolars, while excluding the third molars. The present study did not find any association between various types of malocclusions and hypodontia.

Pharmacological sphingosine-1 phosphate receptor 1 targeting in cigarette smoke-induced emphysema in mice.

Primarily caused by chronic cigarette smoking (CS), emphysema is characterized by loss of alveolar cells comprising lung units involved in gas exchange and inflammation that culminate in airspace enlargement. Dysregulation of sphingolipid metabolism with increases of ceramide relative to sphingosine-1 phosphate (S1P) signaling has been shown to cause lung cell apoptosis and is emerging as a potential therapeutic target in emphysema. We sought to determine the impact of augmenting S1P signaling via S1P receptor 1 (S1P1) in a mouse model of CS-induced emphysema. DBA2 mice were exposed to CS for 4 or 6 mo and treated with pharmacological agonists of S1P1: phosphonated FTY720 (FTY720-1S and 2S analogs; 0.01-1.0 mg/kg) or GSK183303A (10 mg/kg). Pharmacological S1P1 agonists ameliorated CS-induced lung parenchymal apoptosis and airspace enlargement as well as loss of body weight. S1P1 agonists had modest inhibitory effects on CS-induced airspace inflammation and lung functional changes measured by Flexivent, improving lung tissue resistance. S1P1 abundance was reduced in chronic CS-conditions and remained decreased after CS-cessation or treatment with FTY720-1S. These results support an important role for S1P-S1P1 axis in maintaining the structural integrity of alveoli during chronic CS exposure and suggest that increasing both S1P1 signaling and abundance may be beneficial to counteract the effects of chronic CS exposure.

A 56-Year-Old Man With Emphysema, Rash, and Arthralgia.

CASE PRESENTATION: A 56-year-old man presented to the pulmonary clinic with dyspnea and hypoxemia on exertion. He was an avid biker and skier who had noticed a significant decrease in high-level physical activity over the past 3 years. He reported dyspnea, desaturations at altitudes higher than 9,000 feet, dry cough, tachycardia, and palpitations with exercise. Review of systems was also notable for gluten-intolerance, Raynaud's phenomenon, recurrent skin lesions and joint swelling, pain, and stiffness in the areas overlying the jaw, wrists, knees, and ankles (after capsaicin exposure). He denied fever, chills, anorexia, weight loss, hair loss, ocular symptoms, jaw claudication, chest pain, or lower extremity swelling. He had a five pack-year smoking history, no history of prematurity, childhood asthma, recurrent infections, or environmental and occupational exposure. Based on pulmonary function tests from an outside provider, he had received a diagnosis of exercise-induced asthma and had been prescribed an albuterol inhaler to use on an as-needed basis, which failed to improve his symptoms. He was later prescribed a mometasone-formoterol inhaler, still with no symptomatic improvement.

Sphingosine 1 Phosphate (S1P) Receptor 1 Is Decreased in Human Lung Microvascular Endothelial Cells of Smokers and Mediates S1P Effect on Autophagy.

Destruction of alveoli by apoptosis induced by cigarette smoke (CS) is a major driver of emphysema pathogenesis. However, when compared to cells isolated from non-smokers, primary human lung microvascular endothelial cells (HLMVECs) isolated from chronic smokers are more resilient when exposed to apoptosis-inducing ceramide. Whether this adaptation restores homeostasis is unknown. To better understand the phenotype of HLMVEC in smokers, we interrogated a major pro-survival pathway supported by sphingosine-1-phosphate (S1P) signaling via S1P receptor 1 (S1P1). Primary HLMVECs from lungs of non-smoker or smoker donors were isolated and studied in culture for up to five passages. S1P1 mRNA and protein abundance were significantly decreased in HLMVECs from smokers compared to non-smokers. S1P1 was also decreased in situ in lungs of mice chronically exposed to CS. Levels of S1P1 expression tended to correlate with those of autophagy markers, and increasing S1P (via S1P lyase knockdown with siRNA) stimulated baseline macroautophagy with lysosomal degradation. In turn, loss of S1P1 (siRNA) inhibited these effects of S1P on HLMVECs autophagy. These findings suggest that the anti-apoptotic phenotype of HLMVECs from smokers may be maladaptive, since it is associated with decreased S1P1 expression that may impair their autophagic response to S1P.

Assessment of Gingival Biotypes in Patients Visiting a Tertiary Care Centre in Eastern Nepal.

BACKGROUND: Evaluation of gingival biotype has become a routine procedure in periodontal examination because the type of gingival biotype can positively or negatively affect the outcome of periodontal, restorative, orthodontic and implant therapy. The aim of the study was to assess the proportion of types of gingival biotypes in patients visiting a tertiary care center in eastern Nepal. METHODS: Two hundred and fifty patients between 25 to 45 years attending the Periodontology and Oral Implantology were assessed. Gingival biotype of the patents was determined with Probe Transparency technique Results: Out 250 patients assessed, 73 patients (approximately 29.2 %) had thin gingival biotype and remaining 177 patients (approximately 70.8 %) had thick gingival biotype. The number of the male with thin biotype was 31 whereas the number of the male with thick biotype was 82. Similarly, out of 137 female, 42 had thin biotype and remaining 95 female had thick biotype. The types of biotypes were not associated with gender (p=0.67). CONCLUSIONS: Thicker gingival biotype was the more common type of gingival biotype in patients attending the tertiary care center of Eastern Nepal. The occurrence of thick gingival biotype was more common in Adivasi Janajati ethnic community compared to Brahmin / Chhetri ethnic community.