Contrasting roles of MERS-CoV and SARS-CoV-2 internal proteins in pathogenesis in mice.

Betacoronaviruses encode an internal (I) gene via an alternative reading frame within the nucleocapsid gene, called ORF8b for Middle-East respiratory syndrome coronavirus (MERS-CoV) and ORF9b for severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2. Previous reports suggested that proteins 8b and 9b are involved in evading multiple innate immune signaling pathways. However, their roles in mediating pathogenesis in infected animals have not been determined. In this study, we abrogated the expression of protein 8b in MERS-CoV and protein 9b in SARS-CoV-2. Using mouse models of MERS-CoV and SARS-CoV-2 infection, we found that MERS-CoV lacking protein 8b expression was more virulent, while SARS-CoV-2 lacking protein 9b expression was attenuated compared with the respective wild-type viruses. Upon further analysis, we detected increased levels of type I interferon and enhanced infiltration of immune cells to the lungs of mice infected with MERS-CoV lacking protein 8b expression. These data suggest that the I protein of MERS-CoV plays a role in limiting pathogenesis while that of SARS-CoV-2 enhances disease severity. IMPORTANCE The function of betacoronavirus internal protein has been relatively understudied. The earliest report on the internal protein of mouse hepatitis virus suggested that the internal protein is a structural protein without significant functions in virus replication and virulence. However, the internal proteins of severe acute respiratory syndrome coronavirus (SARS-CoV), Middle-East respiratory syndrome coronavirus, and SARS-CoV-2 have been shown to evade immune responses. Despite the reported functions of the internal protein in these highly pathogenic human coronaviruses, its role in mediating pathogenesis in experimentally infected animals has not been characterized. Our data indicated that despite the similar genomic location and expression strategy of these internal proteins, their effects on virulence are vastly different and virus specific, highlighting the complexity between host-virus interaction and disease outcome.

Arg-tRNA synthetase links inflammatory metabolism to RNA splicing and nuclear trafficking via SRRM2.

Cells respond to perturbations such as inflammation by sensing changes in metabolite levels. Especially prominent is arginine, which has known connections to the inflammatory response. Aminoacyl-tRNA synthetases, enzymes that catalyse the first step of protein synthesis, can also mediate cell signalling. Here we show that depletion of arginine during inflammation decreased levels of nuclear-localized arginyl-tRNA synthetase (ArgRS). Surprisingly, we found that nuclear ArgRS interacts and co-localizes with serine/arginine repetitive matrix protein 2 (SRRM2), a spliceosomal and nuclear speckle protein, and that decreased levels of nuclear ArgRS correlated with changes in condensate-like nuclear trafficking of SRRM2 and splice-site usage in certain genes. These splice-site usage changes cumulated in the synthesis of different protein isoforms that altered cellular metabolism and peptide presentation to immune cells. Our findings uncover a mechanism whereby an aminoacyl-tRNA synthetase cognate to a key amino acid that is metabolically controlled during inflammation modulates the splicing machinery.

A Novel Macrophage-Activating Gel Improves Healing and Skin Quality After CO2 Laser Resurfacing of the Chest.

BACKGROUND: After laser resurfacing, it is imperative that an appropriate postoperative regimen is followed for optimal wound healing. There is currently no consensus about which agents should be used. OBJECTIVE: To evaluate the safety and efficacy of a novel macrophage-activating gel in a Phase 2B trial to be used after fractionated ablative laser resurfacing of the chest. MATERIALS AND METHODS: Forty-two adults who received fractionated CO2 laser resurfacing of the chest were randomized (active or placebo) for 5 consecutive days after procedure. Skin quality at baseline and follow-up was assessed by a blinded evaluator using the Fitzpatrick-Goldman Wrinkle Scale. Subject satisfaction with skin healing and quality was also assessed. RESULTS: At 28 days according to the Fitzpatrick-Goldman Wrinkle Scale, 85% of subjects achieved an improvement of at least 33% for the active group versus 50% in the placebo group (absolute difference 35%; p = .04). Similarly, 75% of subjects achieved an improvement score of at least 33% in elastosis in the active group versus 35% in the placebo group at 28 days (40% absolute difference; p = .011). CONCLUSION: This study confirms the potent effects of the novel macrophage-activating gel for optimization of skin healing and quality after laser resurfacing of the chest.

Serious Adverse Events With Injectable Fillers: Retrospective Analysis of 7,659 Patient Outcomes.

BACKGROUND: In total, 2.7 million injectable filler treatments were performed in 2019 in the United States. Although generally considered to be a safe treatment modality, adverse events may occur in rare situations. OBJECTIVE: Analyze serious adverse events from injectable filler treatments, including infections, cutaneous necrosis, blindness, or delayed-onset nodule formation, spanning 11 years for 3 board-certified dermatologists and review their incidence, management, and outcomes. MATERIALS AND METHODS: A retrospective analysis was performed of injectable filler treatments spanning 11 years at a multipractitioner outpatient clinic. Serious adverse events were identified, and treatment measures were documented. A literature search was performed to determine recent trends and outcomes for comparison. RESULTS: Between January 2009 and August 2020, 18,013 mL of injectable filler was administered to 7,659 patients. Of the 18,013 mL administered, 74.1% comprised hyaluronic acid derivatives, 19.19% poly-l-lactic acid, and 6.71% calcium hydroxylapatite. Four serious adverse events were identified. Three events were delayed-onset skin nodule formation. One adverse event was related to vascular compromise and subsequent cutaneous necrosis. After appropriate treatment, all adverse events resolved without significant long-term sequelae. CONCLUSION: Serious adverse events associated with injectable fillers, when performed by board-certified dermatologists, are extremely rare and can be successfully managed with appropriate treatment.

A Randomized, Evaluator-Blind, Split-Face Study Evaluating the Safety and Efficacy of Calcium Hydroxylapatite for Jawline Augmentation.

INTRODUCTION: Jawline augmentation with calcium hydroxylapatite has not yet been evaluated in a prospective study with a split-face design. This study aims to perform the first randomized controlled, split-face study on the efficacy and safety of calcium hydroxylapatite for jawline augmentation using the needle and cannula technique. OBJECTIVE: To perform the first randomized controlled, split-face study on the efficacy and safety of calcium hydroxylapatite for jawline augmentation using the needle and cannula technique. MATERIALS AND METHODS: This is a single-site, randomized, evaluator-blind trial enrolling a total of 10 healthy subjects with at least Grade 1 (mild) on a 4-point Jawline Scale. One side of the face was randomized to receive 1 to 2 syringes of calcium hydroxylapatite with lidocaine (total of 3 mL) for correction of wrinkles and folds along the jawline using both the cannula and needle method, and a balancing treatment will be performed 1 month later. Blinded investigator and subject evaluations will be performed immediately after treatment and at the 30-, 60-, and 90-day visits. RESULTS: Ten subjects were enrolled and completed the trial. There was a improvement in the degree of wrinkling and skin sagging in the 4-point Jawline Scale, with an average of a 1.3-point improvement in the scale on the day of treatment and at the Day 30 visit, which remained improved greater than baseline after 3 months as graded by blinded investigators. The Clinician Global Aesthetic Improvement Score for the treated side versus control, as assessed by blinded investigators, demonstrated a improvement with a 2.3-point improvement on the 5-point scale, and by the final visit on Day 90, most patients had a much improved appearance from baseline. CONCLUSION: This study demonstrates that calcium hydroxylapatite is effective and safe for restoration and augmentation of the jawline using the unique needle and cannula technique.

Review of Post-laser-resurfacing Topical Agents for Improved Healing and Cosmesis.

BACKGROUND: Laser resurfacing produces a controlled skin injury, resulting in a wound healing response. This wound healing response allows for collagen remodeling, which improves skin texture and tone. Topical agents are often employed following laser treatments to facilitate recovery. The introduction of newer small-molecule technologies allow for improved recovery and cosmesis. OBJECTIVE: We sought to perform a critical review of the safety and efficacy of newer small-molecule technologies employed following laser resurfacing. METHODS: We performed a PubMed search of the generic name of the following topicals and included literature relevant to laser procedures, with an emphasis on laser resurfacing: thermal spring water, conjugated linolenic acid, vitamin C/vitamin E/ferulic acid serum, tripeptide/hexapeptide technology-containing products, growth factor serum and gel, recombinant human epidermal growth factor ointment and gel, red deer umbilical cord lining mesenchymal stem cell extract cream and serum, silicone-based gel, and microparticulate (1-3, 1-6 beta-glucan) gel. RESULTS: Our search of the PubMed database yielded 62 results, out of which 17 clinical studies were included in this publication. The majority of aforementioned topicals show promise in terms of improving post-resurfacing recovery or cosmesis. CONCLUSION: Clinical data regarding these agents is limited by the number and quality of studies. It is therefore challenging to propose a recommendation supporting any particular topical. We provide our own provider-specific post-laser resurfacing protocols to offer insight regarding new small-molecule technologies.

Human airway cells prevent SARS-CoV-2 multibasic cleavage site cell culture adaptation.

Virus propagation methods generally use transformed cell lines to grow viruses from clinical specimens, which may force viruses to rapidly adapt to cell culture conditions, a process facilitated by high viral mutation rates. Upon propagation in VeroE6 cells, SARS-CoV-2 may mutate or delete the multibasic cleavage site (MBCS) in the spike protein. Previously, we showed that the MBCS facilitates serine protease-mediated entry into human airway cells (Mykytyn et al., 2021). Here, we report that propagating SARS-CoV-2 on the human airway cell line Calu-3 - that expresses serine proteases - prevents cell culture adaptations in the MBCS and directly adjacent to the MBCS (S686G). Similar results were obtained using a human airway organoid-based culture system for SARS-CoV-2 propagation. Thus, in-depth knowledge on the biology of a virus can be used to establish methods to prevent cell culture adaptation.

Novel Application of the 730 and 785 nm Picosecond Titanium Sapphire Lasers for the Treatment of Nevus of Ota.

BACKGROUND AND OBJECTIVES: Nevus of Ota is a benign dermal melanocytosis that may pose significant psychosocial distress to patients. Q-switched nanosecond lasers have traditionally been considered the first-line treatment but pain, bleeding, and postinflammatory pigmentary alteration are common adverse effects. Picosecond devices have been increasingly used to treat nevus of Ota with promising results. We present two cases demonstrating novel applications of the 730 and 785 nm picosecond titanium sapphire lasers for the treatment of nevus of Ota in two patients with types III and IV skin. STUDY DESIGN/MATERIALS AND METHODS: A 730 and 785 nm picosecond titanium sapphire laser with pulse durations of 250 and 300 picoseconds, respectively, were used to treat two cases of nevus of Ota. Four to seven treatment sessions were conducted at monthly intervals, and follow-up evaluation was performed 1-3 months following the final treatment session. RESULTS: Both cases demonstrated greater than 75% clearance following treatment. There were no adverse events or pigmentary alteration noted as a result of picosecond titanium sapphire laser treatment. CONCLUSIONS: The 730 and 785 nm picosecond titanium sapphire lasers are safe and effective for the treatment of nevus of Ota. Lasers Surg. Med. 00:00-00, 2021. © 2021 Wiley Periodicals LLC.

Intense Pulsed Light: A Methodical Approach to Understanding Clinical Endpoints.

BACKGROUND: Intense Pulsed Light (IPL) is a non-coherent polychromatic broadband filtered flashlamp that emits light in the spectrum of approximately 400–1200 nm. Its effects on photorejuvenation are well documented. The goal of this study is to help practitioners better conceptualize and fine tune IPL device settings in order to produce the most effective and safest clinical outcome. MATERIALS/METHODS: This was a prospective study testing several filters (515 nm; 560 nm; 590 nm and 530–650; 900–1200 nm vascular filter), fluences, pulse durations, and pulse numbers (ie, multiple sequence pulsing or MSP) with a new IPL system. RESULTS: Post-procedure erythema response was more pronounced with increasing fluence, decreasing wavelength, fewer pulses and shorter pulse duration. The exception was the 515 nm filter with regard to pulse duration, which was observed to have a more pronounced response with longer pulse durations. The overall clinical outcome at the 4-week follow-up visit demonstrated greatest improvement in erythema and pigmentation using the 515 nm filter on a Fitzpatrick Skin Type III individual. CONCLUSION: Greatest clinical endpoint response at 4-week follow-up was observed with more robust initial responses. This was most apparent at higher fluence levels and fewer pulse counts. However, when the IPL is pushed to aggressive parameters, there is risk of hypopigmentation and hair loss as seen in this case study. Skin type is an important consideration when using IPL and MSP adds to its safety profile. J Drugs Dermatol. 2021;20(2):203-207. doi:10.36849/JDD.5638.

A Systematic Review of Picosecond Laser in Dermatology: Evidence and Recommendations.

BACKGROUND AND OBJECTIVES: The use of picosecond laser in dermatology was originally focused on optimizing the removal of unwanted tattoos. Subsequent advances in this technology have broadened its clinical indications to include treatment of benign pigmented lesions, photodamage, melasma, and scar revision. In this systematic review, evidence-based recommendations are developed for the use of picosecond laser in dermatology. STUDY DESIGN/MATERIALS AND METHODS: A comprehensive search of the English language literature was performed up to and including November 2019. Relevant citations were individually evaluated, synthesized, and categorized based on the Level of Evidence. With the addition of the authors' combined clinical experience, clinical recommendations were developed. RESULTS: After application of inclusion and exclusion criteria, a total of 77 unique studies were evaluated. Treatment of benign pigmented lesions was associated with level I-IV evidence; rejuvenation was associated with level II evidence; melasma was associated with level II evidence; scar revision was associated with level II-III evidence; tattoo removal was associated with level I evidence. CONCLUSION: Picosecond laser is a safe and effective treatment modality for an increasing range of dermatologic indications. Further development of this technology is warranted. Lasers Surg. Med. © 2020 Wiley Periodicals, Inc.

A Randomized, Split-Face, Double-Blind Comparison Trial Between Fractionated Frequency-Doubled 1064/532 nm Picosecond Nd:YAG Laser and Fractionated 1927 nm Thulium Fiber Laser for Facial Photorejuvenation.

BACKGROUND AND OBJECTIVE: Fractionated lasers are a popular therapeutic option for facial photorejuvenation. In this study, we compare the safety, tolerability, and efficacy of a fractionated frequency-doubled 1064/532 nm picosecond Nd:YAG fractionated picosecond laser (FPL) versus a fractionated 1927 nm thulium fiber laser (TFL) for facial rejuvenation. STUDY DESIGN/MATERIALS AND METHODS: This was a double-blind, randomized, split-face comparison study involving 20 subjects. Facial halves were randomized to receive either FPL or TFL treatment. Three treatments were delivered at 1-month intervals. Subjects were followed up for 1, 3, and 6 months post-final treatment session and evaluated by blinded, non-treating investigators for dyspigmentation, erythema, keratosis, texture, and rhytids on a standardized scale. Subjects also recorded a quantitative daily diary rating healing progress for 14 days after every treatment session. RESULTS: Statistically significant improvements in elastosis, erythema, keratosis, dyschromia, and skin texture were noted in both treatment groups. There were no significant differences detected in clinical efficacy between the two groups. Subject daily dairies revealed statistically significant differences in tolerability during the immediate 14-day post-operative recovery period. The facial half treated with FPL displayed significantly less redness on days 3 and 4; significantly less swelling on day 5; significantly less crusting on days 1 through 9; significantly less peeling on days 3 through 9; and significantly less itch on days 4 and 7. There were no unexpected adverse effects observed. CONCLUSION: Both FPL and TFL are safe and effective treatment options for facial rejuvenation. FPL may be associated with significantly less downtime. Lasers Surg. Med. © 2020 Wiley Periodicals, Inc.

Safety and Efficacy of a Novel 730 nm Picosecond Titanium Sapphire Laser for the Treatment of Benign Pigmented Lesions.

BACKGROUND AND OBJECTIVES: Picosecond lasers in dermatology were originally focused on optimizing the removal of unwanted tattoos. Subsequent advances in this technology have expanded its clinical indications to include treatment of benign pigmented lesions, photodamage, melasma, and scar revision. In this retrospective review, we evaluate a novel 730 nm picosecond titanium sapphire laser in treating benign pigmented lesions. STUDY DESIGN/MATERIALS AND METHODS: This is a retrospective review of all patients who presented to our institution between December 2019 and March 2020 for treatment of their benign pigmented lesions with a 730 nm picosecond titanium sapphire laser. All Fitzpatrick skin types (I-VI) were included. Absolute and relative evaluations were conducted by two blinded board-certified dermatologists using high-resolution photographic images. RESULTS: Twenty-two of 64 patients satisfied inclusion and exclusion criteria. Patients received 1.1 ± 0.3 treatment sessions. The absolute average pigmentation score prior to treatment was 2.04 ± 0.7 versus 1.39 ± 0.6 after treatment (P < 0.05). Improvement in pigmentation was observed in 86% of the patients, while 3% had no improvement and 11% had worsening of pigmentation. No other adverse events were observed. Downtime consisted of 1-2 days of mild edema and erythema followed by 3-5 days of mild pigment darkening and superficial crust. CONCLUSION: The novel 730 nm picosecond titanium sapphire laser is a safe and effective treatment for benign pigmented lesions. Future prospective randomized control studies would be beneficial to further clarify its role in the treatment of benign pigmentation. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

Successful Treatment of Scalp Micropigmentation With 1064 nm Picosecond Nd:YAG Laser.

BACKGROUND AND OBJECTIVES: Scalp micropigmentation (SMP) is a useful technique that applies permanent camouflage for hair loss and scarring. However, suboptimal outcomes necessitate removal. To date, there have been no reports of SMP removal with a picosecond laser. In this report, we demonstrate the safety and efficacy of 1064 nm picosecond neodymium yttrium aluminum garnet (Nd:YAG) laser for the removal of SMP. STUDY DESIGN/MATERIALS AND METHODS: This was a case series of four patients who presented with undesirable SMP. Following induction of adequate cutaneous anesthesia, a 1064 nm picosecond Nd:YAG laser was applied to all surface areas with SMP. Post-operative care consisted of 1% dimethicone ointment applied twice daily to the affected areas for 3-7 days. Patients were evaluated after 3-4 weeks and additional treatment sessions were undertaken if necessary. RESULTS: Clearance of SMP was achieved within 1-3 treatment sessions in all cases. There were no unexpected adverse events. CONCLUSION: Picosecond Nd:YAG laser is an excellent treatment option for undesirable SMP. It is safe and effective and typically requires fewer treatment sessions than traditional tattoo removal. Lasers Surg. Med. © 2020 Wiley Periodicals LLC.

A Prospective Double-blind, Placebo-controlled Clinical Trial Evaluating the Efficacy of a Novel Combination of Hyaluronic Acid Serum and Antioxidant Cream for Rejuvenation of the Aging Neck.

BACKGROUND: The neck is one of the most common areas affected by the aging process. A novel two product combination system composed of a serum and cream with hyaluronic acid and multiple strong antioxidants were investigated to determine their efficacy and safety in neck rejuvenation. OBJECTIVE: The objective of this prospective, randomized, double-blind, placebo-controlled clinical trial was to assess the efficacy and safety of a novel serum containing fractionated hyaluronic acid, peptides, and antioxidants for photodamage of the neck. METHODS: This was an institutional review board (IRB)-approved, randomized, double blind, placebo-controlled clinical trial involving 31 healthy subjects with moderate-to-severe neck wrinkling corresponding to at least a Grade 2 in wrinkles and score of 4 in elastosis on the Fitzpatrick-Goldman Wrinkle Scale. Twenty subjects were randomized to receive the active cream and serum system, while 11 subjects were randomized to receive the vehicles alone in serum and cream format for a course of two months. RESULTS: Both active and placebo cream and serum showed improvement of wrinkles, laxity, pigmentation, erythema, dryness, and texture of the skin, and high patient satisfaction scores. Histology of one of the active serums and cream samples revealed improvement in the quality of papillary dermal collagen and increase in the number of elastic fibers in the upper dermis after treatment. CONCLUSION: Our prospective, randomized controlled trial showed that the novel serum and cream showed improvement in skin aging on the neck, was well-tolerated by patients, and had a high degree of patient satisfaction.

Identification of protein-protected mRNA fragments and structured excised intron RNAs in human plasma by TGIRT-seq peak calling.

Human plasma contains > 40,000 different coding and non-coding RNAs that are potential biomarkers for human diseases. Here, we used thermostable group II intron reverse transcriptase sequencing (TGIRT-seq) combined with peak calling to simultaneously profile all RNA biotypes in apheresis-prepared human plasma pooled from healthy individuals. Extending previous TGIRT-seq analysis, we found that human plasma contains largely fragmented mRNAs from > 19,000 protein-coding genes, abundant full-length, mature tRNAs and other structured small non-coding RNAs, and less abundant tRNA fragments and mature and pre-miRNAs. Many of the mRNA fragments identified by peak calling correspond to annotated protein-binding sites and/or have stable predicted secondary structures that could afford protection from plasma nucleases. Peak calling also identified novel repeat RNAs, miRNA-sized RNAs, and putatively structured intron RNAs of potential biological, evolutionary, and biomarker significance, including a family of full-length excised intron RNAs, subsets of which correspond to mirtron pre-miRNAs or agotrons.

The Efficacy of Pulsed Dye Laser Pretreated With or Without Local Anesthetic on Patients Presenting With Erythema of Face, Neck, Chest, and Extremities.

BACKGROUND AND OBJECTIVES: Erythema is one of the most common cosmetic concerns and usually responds well to pulsed dye laser (PDL) treatment. As this laser can cause significant discomfort, topical anesthesia is sometimes offered. However, it is still uncertain whether topical anesthetics can affect the outcome of the laser therapy. We performed a retrospective single site study to compare the efficacy of PDL for the treatment of erythema in patients with and without pretreatment with topical anesthetic. STUDY DESIGN/MATERIALS AND METHODS: A chart review was performed and patients who presented with erythema of face, neck, chest, and extremities pretreated with topical anesthesia (23% lidocaine/7% tetracaine ointment or 7% lidocaine/7% tetracaine ointment) undergoing PDL were reviewed and compared with another group without anesthesia. Two blinded dermatologists evaluated the postlaser procedure photographs and gave an assessment compared with baseline. RESULTS: A total of 69 patient charts were reviewed. The erythema resulted from various skin conditions including telangiectasia, cherry angioma, striae, and rosacea. The mean improvement was 2.2581 in the anesthesia group and 2.2632 in the nonanesthesia group. There was no significant difference between both groups as confirmed by a noninferiority test. CONCLUSIONS: Topical anesthesia with lidocaine and tetracaine ointment do not interfere with the efficacy of the PDL. Since pain management is essential for any cosmetic procedure, the application of a local anesthetic will enhance patient comfort and satisfaction during treatment with PDL. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc.

Preventing an Antigenically Disruptive Mutation in Egg-Based H3N2 Seasonal Influenza Vaccines by Mutational Incompatibility.

Egg-based seasonal influenza vaccines are the major preventive countermeasure against influenza virus. However, their effectiveness can be compromised when antigenic changes arise from egg-adaptive mutations on influenza hemagglutinin (HA). The L194P mutation is commonly observed in egg-based H3N2 vaccine seed strains and significantly alters HA antigenicity. An approach to prevent L194P would therefore be beneficial. We show that emergence of L194P during egg passaging can be impeded by preexistence of a G186V mutation, revealing strong incompatibility between these mutations. X-ray structures illustrate that individual G186V and L194P mutations have opposing effects on the HA receptor-binding site (RBS), and when both G186V and L194P are present, the RBS is severely disrupted. Importantly, wild-type HA antigenicity is maintained with G186V, but not L194P. Our results demonstrate that these epistatic interactions can be used to prevent the emergence of mutations that adversely alter antigenicity during egg adaptation.