Thg1 family 3'-5' RNA polymerases as tools for targeted RNA synthesis.

Members of the 3'-5' RNA polymerase family, comprised of tRNAHis guanylyltransferase (Thg1) and Thg1-like proteins (TLPs), catalyze templated synthesis of RNA in the reverse direction to all other known 5'-3' RNA and DNA polymerases. Discovery of enzymes capable of this reaction raised the possibility of exploiting 3'-5' polymerases for post-transcriptional incorporation of nucleotides to the 5'-end of nucleic acids without ligation, and instead by templated polymerase addition. To date, studies of these enzymes have focused on nucleotide addition to highly structured RNAs, such as tRNA and other non-coding RNA. Consequently, general principles of RNA substrate recognition and nucleotide preferences that might enable broader application of 3'-5' polymerases have not been elucidated. Here, we investigated the feasibility of using Thg1 or TLPs for multiple nucleotide incorporation to the 5'-end of a short duplex RNA substrate, using a templating RNA oligonucleotide provided in trans to guide 5'-end addition of specific sequences. Using optimized assay conditions, we demonstrated a remarkable capacity of certain TLPs to accommodate short RNA substrate-template duplexes of varying lengths with significantly high affinity, resulting in the ability to incorporate a desired nucleotide sequence of up to 8 bases to 5'-ends of the model RNA substrates in a template-dependent manner. This work has further advanced our goals to develop this atypical enzyme family as a versatile nucleic acid 5'-end labeling tool.

Impact of Formulation and Microneedle Length on Transdermal Metronidazole Permeation through Microneedle-Treated Skin.

OBJECTIVE: This study aimed to determine the impact of formulation (gel vs cream) and microneedle characteristics (length, number) on permeation of metronidazole through excised microneedle-treated skin. The long-term goal is to apply these results towards a pharmacokinetic study in human subjects with diverse skin types, using in vitro flux data to determine dosing conditions and ultimately establish in vitro-in vivo correlations. METHODS: Metronidazole release from 0.75% gel and cream was quantified with flow-through diffusion cells, using a cellulose membrane. Excised porcine skin was treated with stainless steel microneedles (500 or 800 µm length), to create 50 or 100 micropores. Metronidazole gel or cream was applied to microneedle-treated skin and replaced every 48 h for up to 7 days. Metronidazole permeation was quantified using HPLC. Intact skin (no microneedle treatment) served as controls. RESULTS: Metronidazole release was faster from the gel vs cream. At 7 days there was no difference between gel vs cream in total metronidazole permeated through intact skin. For both formulations, metronidazole permeation was significantly higher (vs intact skin) following microneedle application, regardless of microneedle length or micropore number. Increasing microneedle length and micropore number enhanced MTZ permeation multiple fold for both gel and cream. The greatest enhancement in total permeation for both formulations was achieved with the 800 µm MN, 100 micropore condition. CONCLUSIONS: Formulation and microneedle conditions both impacted metronidazole permeation. These data will be used to estimate in vivo serum concentrations after applying metronidazole to microneedle-treated skin in humans.

How to assess nonresponsiveness to vasodilator stress.

Myocardial perfusion imaging (MPI) is a powerful tool for the functional assessment of ischemia in patients with suspected or known coronary artery disease (CAD). Given that the diagnostic accuracy and prognostic value of MPI and post-test management are highly dependent on achieving an adequate stress vasodilatory response, it is critical to identify those who may not have adequately responded to vasodilator pharmacological stress agents such as adenosine, dipyridamole, and regadenoson. Caffeine, a potent inhibitor of the adenosine receptor, is a compound that can affect vasodilatory hemodynamics, result in false negative studies, and potentially alter management in cases of inaccurate test results. Vasodilator non-responsiveness can be suspected by examining hemodynamics, quantitative positron emission tomography (PET) metrics such as myocardial flow reserve (MFR), and splenic response to stress. Quantitative MFR values of 1-1.2 should raise suspicion for nonresponsiveness in the setting of normal perfusion, along with the absence of a splenic switch off. Newer metrics, such as splenic response ratio, can be used to aid in the identification of potential nonresponders to pharmacologic vasodilators.

Surgical outcomes and revision rates for velopharyngeal insufficiency (VPI) in syndromic and non-syndromic children: A systematic review and meta-analysis.

PURPOSE: To evaluate pre- and post-operative resonance, surgical technique, revision rate, and revision indication among syndromic and non-syndromic children with velopharyngeal insufficiency (VPI). MATERIALS AND METHODS: A systematic review was conducted through July 2022. Children surgically treated for VPI were included. A meta-analysis of single means, proportions, comparison of proportions, and mean differences with 95 % confidence interval [CI] was conducted. RESULTS: Twenty-three articles (n = 1437) were included in the analysis. The most common surgery was Sphincter Pharyngoplasty (SP), 62.6 % [31.3-88.9] for syndromic and 76.3 % [37.5-98.9] for non-syndromic children. Among all surgical techniques, for syndromic and non-syndromic children, 54.8 % [30.9-77.5] and 73.9 % [61.3-84.6] obtained normal resonance post-operatively, respectively. Syndromic patients obtained normal resonance post-operatively in 83.3 % [57.7-96.6] of Combined Furlow Palatoplasty and Sphincter Pharyngoplasty (CPSP), 72.6 % [54.5-87.5] of Pharyngeal Flap (PF), and 45.1 % [13.2-79.8] of Sphincter Pharyngoplasty (SP) surgeries. Non-syndromic patients obtained normal resonance post-operatively in 79.2 % [66.4-88.8] of PF and 75.2 % [61.8-86.5] of SP surgeries. The revision rate for syndromic and non-syndromic patients was 19.9 % [15.0-25.6] and 11.3 % [5.8-18.3], respectively. The difference was statistically significant, 8.6 % [2.9-15.0, p = 0.003]. Syndromic patients who underwent PF were least likely to undergo revision surgery as compared to SP and CPSP, 7.7 % [2.3-17.9] vs. 23.7 % [15.5-33.1] and 15.3 % [2.8-40.7], respectively. CONCLUSIONS: Syndromic children had higher revision rates and were significantly less likely to obtain normal resonance following primary surgery than non-syndromic patients. Among syndromic children, PF and CPSP have been shown to improve resonance and reduce revision rates more so than SP alone.

Implant-Based Gluteal Augmentation: Comparing Complications Between Single- and Double-Incision Techniques.

BACKGROUND: Gluteal implants are gaining popularity as an alternative to fat grafting for gluteal augmentation. This study aims to compare complication rates between single- and double-incision techniques for implant placement in gluteal augmentation from a single surgeon's experience. METHODS: Retrospective analysis of a single surgeon's implant-based gluteal augmentations was conducted between October 2018 and August 2022. Consecutive patient cases were reviewed at the beginning and end of the designated period to compare both incision techniques after switching from the single- to double-incision technique in September 2020. Demographics and postoperative complications were compared. Statistical comparisons were made using the independent sample t-test for quantitative variables and the Fisher's exact test for categorical variables. RESULTS: The study included 134 single-incision patients (247 implants) and 47 double-incision patients (91 implants). Baseline demographic and clinical characteristics between the two groups were similar. Overall complication rate per implant was higher in the single-incision group (16.6% vs. 6.6%, p = 0.02). The most common complication per implant was delayed wound healing (single incision: 9.7%, double incision: 5.5%). The odds ratio of developing at least one complication in the double compared to the single-incision group was 0.44 (95% CI: 0.19, 0.92). CONCLUSIONS: The double incision technique demonstrates a lower complication rate than the single-incision technique for implant-based gluteal augmentation. The most common complication in both groups was delayed wound healing which responded in most patients to conservative wound care. These results support using the double-incision technique for implant-based gluteal augmentation. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Cleft Lip and/or Palate Repair in Children With Hypopituitarism: Analysis of the Kids' Inpatient Database.

OBJECTIVE: Children with hypopituitarism (CwHP) can present with orofacial clefting, frequently in the setting of multiple midline anomalies. Hypopituitarism (HP) can complicate medical and surgical care; the perioperative risk in CwHP during the traditionally lower risk cleft lip and/or palate (CL/P) repair is not well described. The objective of this study is to examine the differences in complications and mortality of CL/P repair in CwHP compared to children without hypopituitarism (CwoHP). DESIGN: A retrospective cross-sectional analysis. SETTING: The 1997 to 2019 Kids' Inpatient Databases (KID). PATIENTS: Children 3 years old and younger who underwent CL/P repair. MAIN OUTCOME MEASURE(S): Complications and mortality. RESULTS: A total of 34 106 weighted cases were analyzed, with 86 having HP. CwHP had a longer length of stay (3.0 days [IQR 2.0-10.0] vs 1.0 day [IQR 1.0-2.0], P < .001) and higher rates of complications and mortality (12.8% vs 2.9%, P < .001) compared to CwoHP. Controlling for demographic factors, CwHP had 6.61 higher odds of complications and mortality than CwoHP (95% CI 3.38-12.94, P < .001). CONCLUSIONS: CwHP can present with a CL/P and other midline defects that can increase the complexity of their care. These data show a significant increase in length of stay, complications, and mortality in CwHP undergoing CL/P repair. Increased multidisciplinary attention and monitoring may be needed for these children peri- and postoperatively, especially if additional comorbidities are present. Further studies on perioperative management in this population are warranted to reduce morbidity and mortality.

Help-Seeking Behaviors in a Community Experiencing Multiple, Concurrent Natural Disasters. Lessons from a Greater Houston Community.

Previous disaster research exploring mental health consequences has been conducted after a single disaster. Community mental health nurses offer critical support in the aftermath of a disaster through complex assessments, psychosocial education and treatment. This research examines help-seeking behaviors in a Greater Houston community struggling with two simultaneous disasters, a historic winter freeze amidst the COVID-19 pandemic. Three Mental Health Crisis clinics, staffed by an interdisciplinary team, offered walk-in crisis services. The study characterizes adults who accessed services and describes those who accepted follow-up, subsidized behavioral health therapy or case management, including populations experiencing health disparities. Statistical analysis included descriptive and inferential statistics. Those who recognized the need for behavioral health care and accessed episodic care differed from those who accepted follow-up care. The population that was more likely to accept follow up care largely consisted of females, individuals without a previously existing mental health condition, those who reported trauma exposure, and young adults of Hispanic/Latino origin. Nurses can leverage female support to inspire male adults to seek care and advocate for increased resources and training to support culturally relevant care. Nurses and the interdisciplinary team need to assess for trauma exposure, as it influences care decisions.

Structural Support for Large to Total Nasal Reconstruction.

When large defects of the nose are present, it is imperative to address all 3 layers: the external skin envelope, the osteocartilaginous support, and the inner mucosal lining. The middle structural framework is the primary factor in determining the overall shape of the nose, in addition to facilitating a functional and patent airway. As such, its reconstruction must be robust enough to provide lasting osteocartilaginous support while minimizing disfiguring bulk. The goal is replacement of missing tissue with grafts of similar strength, size, and shape. This article will review approaches to the reconstruction of structural support in large nasal defects.

Surgical Opportunism for Hand Reconstruction Following a Mutilating Injury: A Case Report.

We report our management of a 53-year-old female who suffered a wood planer hand-mutilating injury with significant dorsal soft tissue loss and partial metacarpophalangeal joint (MCPJ) amputations of the thumb, index, and middle fingers. The middle finger was deconstructed for "spare parts" and a vascularized osteochondral graft was utilized to reconstruct the metacarpal articular surface of the index finger proximal phalanx, allowing the pedicled transposition of the index finger to the third metacarpal. The middle finger's distal interphalangeal joint was transplanted non-vascularly to recreate the thumb MCPJ and the elevation of a middle finger fillet flap allowed dorsal wound coverage. The patient did well initially but required ulnar collateral ligament reconstruction with a palmaris longus tendon graft following MCPJ instability 10 months postoperatively. Nonetheless, she progressively regained thumb opposition and pinch grip and continues to have successful aesthetic and functional outcomes six years postoperatively, supporting the efficacy of non-vascularized joint transfers when vascularized options are superfluous or unavailable.

Suture fixation of traumatic rib fracture flail segment with Klebsiella osteomyelitis.

We report the case of a 71-year-old male who initially presented with urosepsis and was found to have a rib fracture of his right 6th rib with a flail segment and an associated abscess. Given the concern for infection, surgical rib fixation with titanium plating was not pursued during the washout of his abscess and instead, he successfully underwent rib fracture stabilization with bilateral suture transfixation. He was continued on a prolonged course of antibiotics for Klebsiella pneumonia osteomyelitis and was discharged uneventfully with optimal pain control and adequate respiratory effort.

High-Grade Pleomorphic Sarcoma in Postauricular Keloid Scar After Surgical Excision and Radiation.

Keloid scars can cause significant morbidity to the patient including substantial cosmetic disfigurement, particularly in the head and neck region. Surgical excision followed by immediate postoperative radiation therapy has been shown to be more effective than single modality treatment. Radiation therapy increases risk for radiation-induced malignancy, though very few cases have been reported in the treatment of keloids. We report the case of a patient with a large postauricular keloid treated with excision and immediate post-operative radiation who developed a high-grade pleomorphic sarcoma in the scar bed. Laryngoscope, 134:3143-3145, 2024.

Influenza A (H1N1) Virus Outbreak in the Districts of Chhattisgarh: A Cross-Sectional Study.

Background The H1N1 flu is a subtype of the influenza A virus, also known as the swine flu. An entirely new strain of the H1N1 virus started sickening people in the 2009-2010 flu season. It was a novel influenza virus combination that can infect humans, pigs, and birds. It was frequently referred to as the "swine flu." The virus may be able to spread for a little while longer in children and individuals with compromised immune systems. Objective The objective is to investigate the outbreaks of H1N1 among young adults in the Bastar District of Chhattisgarh. Methods Collection of the blood samples of 342 individuals between December 2015 and November 2017 was done. Thirty-one cases of Influenza A (H1N1) PDM09 virus infection were identified and confirmed. The molecular relationship between viruses is identified by the real-time polymerase chain reaction (RT-PCR) method. Result The majority of samples (n=13) were sourced from Raipur Medical College, followed by contributions from Durg District Hospital (n=5), Raigarh Medical College (n=4), Rajnandgaon District Hospital (n=3), Jagdalpur Medical College (n=2), Bilaspur Medical College (n=2), and smaller contributions from Dhamtari District Hospital and Gariyabandh Primary Health Care. Among these, 31 samples tested positive for Influenza A (H1N1) PDM 2009 virus, with a slightly higher prevalence among 19 female patients. Age-wise distribution revealed higher proportions of positive cases in the age groups of 0-10 years, 31-40 years, and 21-30 years. In the molecular analysis, 154 samples showed no target amplification, while 125 samples exhibited amplification of only Influenza A without subtype (H1) amplification. Remarkably, 31 patients who tested positive for Influenza A (H1N1) died from the virus; most of the deaths were in children under five and middle-aged adults. Conclusion The detection of Influenza A (H1N1) PDM 2009 virus, especially among females, indicates its persistent circulation. Positive cases were prevalent among younger and middle-aged individuals. Molecular analysis showed subtype variations, with significant fatalities observed in children under five and middle-aged adults, emphasizing the severity of the virus across different age groups. It is advised that in order to keep Indian influenza surveillance up to date and robust, more epidemiological data should be gathered, along with information on risk factors like immunization status, hospitalization, and mortality rates should be estimated, and influenza case subtyping should be improved.

Plastic Surgery Tourism: Complications, Costs, and Unnecessary Spending?

Background: The liability of plastic surgery tourism in patient health and postoperative resource allocation is significant. Procedures completed within the context of medical tourism often lack rigorous quality assurance and provide limited preoperative evaluation or postoperative care. Not only does this jeopardize the patient's well-being, but it also increases the financial burden and redirects invaluable resources domestically through often unnecessary diagnostic tests and hospitalizations. Objectives: This manuscript will examine the complications and associated costs following plastic surgery tourism and highlight unnecessary expenses for patients with outpatient complications. Methods: A retrospective review was conducted of all patients 18 years or older who underwent destination surgery and were seen within 1 year postoperatively in consultation with plastic surgery at our health system between January 11, 2015 and January 7, 2022. Patient admissions were reviewed and deemed necessary or unnecessary after review by 2 physicians. Results: The inclusion criteria were met by 41 patients, of whom hospitalization was deemed necessary in 28 patients vs unnecessary in 13 patients. The most common procedures included abdominoplasty, liposuction, breast augmentation, and "Brazilian butt lift." The most common complications were seroma and infection. Patients deemed to have a necessary admission often required at least 1 operation, were more likely to need intravenous antibiotics, were less likely to have the diagnosis of "pain," necessitated a longer hospitalization, and incurred a higher cost. The total financial burden was $523,272 for all 41 patients. Conclusions: Plastic surgery tourism poses substantial health risks, the morbidities are expensive, and it strains hospital resources.

Endovascular covered stent graft repair by through-and-through snaring of an acute traumatic femoral-femoral arteriovenous fistula following a gunshot wound.

Arteriovenous fistula (AVF) formation after penetrating injury underscores a rare and challenging complication of vascular trauma. Traumatic AVFs have various clinical presentations and reported methods of repair. Although open surgical repair is the most frequently used method of repair, the advancement of endovascular techniques has been increasingly used during the past 3 decades. We report a case of an acute traumatic AVF of the superficial femoral artery and superficial femoral vein from a gunshot wound repaired with a unique endovascular technique involving snaring to establish through-and-through access to allow deployment of a covered stent graft.

Alternative Linkage Chemistries in the Chemoenzymatic Synthesis of Microviridin-Based Cyclic Peptides.

Engineering biosynthetic pathways to ribosomally synthesized and post-translationally modified peptides (RiPPs) offers several advantages for both in vivo and in vitro applications. Here we probe the ability of peptide cyclases to generate trimacrocycle microviridin analogs with non-native cross-links. The results demonstrate that diverse chemistries are tolerated by macrocyclases in the ATP-grasp family and allow for the construction of unique cyclic peptide architectures that retain protease inhibition activity. In addition, cocomplex structures of analogs bound to a model protease were determined, illustrating how changes in functional groups maintain peptide conformation and target binding.

When Blood Is Thicker Than Water: A Case of Acute Pancreatitis Secondary to Familial Hypertriglyceridemia.

Hypertriglyceridemia is one of the major causes of acute pancreatitis in addition to gallstones and alcohol use. These etiologies are often associated with underlying comorbidities. Acute pancreatitis secondary to hypertriglyceridemia is associated with an increase in clinical severity and further complications. We present a case of a 56-year-old man with a past medical history of hypertension, diabetes mellitus, and familial hypertriglyceridemia who was diagnosed with acute pancreatitis secondary to hypertriglyceridemia. The patient presented with 9/10 pressure across the abdomen radiating to the sternum. Labs revealed elevated triglyceride count > 8000 mg/dL and cholesterol > 705 mg/dL. Abdominal CT showed fat stranding along the anterior aspect of the pancreatic head. The patient was managed with IV fluids, nil per os (NPO), and statin management for hypertriglyceridemia. Seven days later, triglycerides decreased to 658 mg/dL, and abdominal pain resolved. This case highlights an unusual presentation of acute pancreatitis and demonstrates the importance of understanding the spectrum of etiologies for this condition.