A scalable and cost-efficient rRNA depletion approach to enrich RNAs for molecular biology investigations.

Transcriptomics analyses play pivotal roles in understanding the complex regulatory networks that govern cellular processes. The abundance of rRNAs, which account for 80%-90% of total RNA in eukaryotes, limits the detection and investigation of other transcripts. While mRNAs and long noncoding RNAs have poly(A) tails that are often used for positive selection, investigations of poly(A)- RNAs, such as circular RNAs, histone mRNAs, and small RNAs, typically require the removal of the abundant rRNAs for enrichment. Current approaches to deplete rRNAs for downstream molecular biology investigations are hampered by restrictive RNA input masses and high costs. To address these challenges, we developed rRNA Removal by RNaseH (rRRR), a method to efficiently deplete rRNAs from a wide range of human, mouse, and rat RNA inputs and of varying qualities at a cost 10- to 20-fold cheaper than other approaches. We used probe-based hybridization and enzymatic digestion to selectively target and remove rRNA molecules while preserving the integrity of non-rRNA transcripts. Comparison of rRRR to two commercially available approaches showed similar rRNA depletion efficiencies and comparable off-target effects. Our developed method provides researchers with a valuable tool for investigating gene expression and regulatory mechanisms across a wide range of biological systems at an affordable price that increases the accessibility for researchers to enter the field, ultimately advancing our understanding of cellular processes.

Static Suspension of the Paralyzed Face Utilizing the Midfacial Corridor: Anatomic Evaluation and Surgical Technique.

BACKGROUND: Fascia lata and tendon grafts are frequently utilized to support the paralyzed midface and to extend muscular reach in McLaughin style, orthodromic temporalis transfers. The grafts are frequently placed in a deep subcutaneous positioning that can lead to the development of a, bowstring deformity in the cheek. This paper describes insertion of tendon grafts into the midfacial corridor collectively formed by the buccal, submasseteric and superficial temporal spaces. METHODS: Over a seven-year period, all patients that underwent insertion of facia lata and tendon grafts in the midfacial corridor were included. Demographic information, perioperative variables and clinical outcomes were collected and analyzed. RESULTS: A total of 22 patients were included with a mean age of 64.3 years (33-86). There were multiple etiologies for the facial weakness including acoustic neuroma (9.1%), Bell's palsy (13.6%), facial nerve schwannoma (9.1%), temporal bone fracture (4.6%) and malignancy (22.7%). Midfacial corridor grafts were utilized in combination with nerve transfers (V-VII and XII-VII) in nine patients, McLaughin style temporalis transfers in 12 and as a standalone procedure in one individual. During the study period, no patients exhibited a tethering, or concave deformity in the midface. Additionally, no impingement, difficulties with mastication, parotitis or hematoma were encountered. One patient developed a postoperative infection, that was successfully managed. CONCLUSION: Placement of tendon or fascia grafts for static support or tunneling of an orthodromic temporalis transfer through the midfacial corridor can be performed rapidly while providing midfacial support and avoiding the creation of visible cutaneous deformities.

Midline Intranasal Dermoid Cyst With Intracranial Extension: Technical Details of a Multidisciplinary Approach to a Rare Condition.

Dermoid cysts are rare, benign neoplasms that develop when ectodermal tissue is ectopically included during neural tube closure. Only 0.7% to 1.8% of dermoid cysts occur intracranially, and these make up only 0.3% of all intracranial tumors. Definitive management of intracranial dermoid cysts is achieved with surgical excision, with the primary goal being removal of the lesion before rupture or infection occurs. The authors report the multidisciplinary management of a 6-month-old patient with a midline intranasal dermoid cyst with intracranial involvement successfully treated with surgical resection. The surgical approach included Neurosurgical access with a bifrontal approach to remove the intracranial portion, then Plastic Surgery accessed and removed the intranasal portion through a transcolumellar and infracartilaginous dissection. The entire cyst was successfully removed without rupture. The patient's postoperative course was without complication.Timely preoperative imaging and diagnosis were crucial to the success of this operation. When considering the possibility of a midline dermoid cyst with intracranial extension, it is essential to obtain proper imaging to distinguish between lesions on the differential diagnosis and to define extent of extension. Once diagnosed, surgical planning requires a multidisciplinary approach. Considerations for the optimal surgical approach to intracranial, intranasal dermoids include maximizing surgical exposure and ensuring an aesthetically pleasing reconstructive outcome.

One-Step Salvage of Infected Prosthetic Breast Reconstructions Using Antibiotic-Impregnated Polymethylmethacrylate Plates and Concurrent Tissue Expander Exchange.

PURPOSE: Periprosthetic infection represents a major complication in breast reconstruction, frequently leading to expander-implant loss. Recent studies report variable success in the salvage of infected breast prostheses through systemic antibiotic therapy and surgical intervention. There is currently no consensus regarding a management algorithm for attempted salvage. The purpose of this pilot study was to evaluate the early outcomes of a protocol using antibiotic-impregnated polymethylmethacrylate (PMMA) implant placement with expander device exchange. METHODS: A retrospective database was queried to identify all patients with infected implant-based breast reconstruction who were treated by the study authors and who underwent attempted salvage under the study protocol. All patients received intravenous antibiotics followed by surgical debridement of the infected pocket, insertion of antibiotic-impregnated PMMA plates and/or beads, device exchange, and postoperative antibiotics. After clinical resolution of infection, tissue expansion was performed with the PMMA implants remaining in situ until exchanged to permanent implants. RESULTS: All patients with infected prosthetic breast reconstructions achieved implant pocket sterilization using this method. At a mean follow-up of 8.2 months (range, 1-19 months), none of these patients have required reoperation for capsular contracture. One patient, while under treatment with prednisone for a rash, developed recurrent infection, which led to explantation of her implant. Two patients underwent radiation therapy while an antibiotic plate and tissue expander were in place, with no observed exposure or infection recurrence. CONCLUSIONS: Sustained local antibiotic delivery using PMMA implants and expander device exchange can successfully salvage an infected breast implant. Perceived benefits include shorter time to completed reconstruction, preserved skin envelope integrity, and possibly improved long-term aesthetic outcomes.

Pharyngoesophageal reconstruction outcomes following 349 cases.

BACKGROUND: Pharyngoesophageal (PE) reconstruction is complex, with a diverse set of reconstructive considerations. This large series examines the impact of various defect characteristics and reconstructive modalities on outcomes. PATIENTS AND METHODS: A retrospective review identified 349 cases from 2000 to 2011. Patients were grouped according to defect extent and location. Groups were compared by comorbidities, flap type, donor and recipient site complications, postoperative diet, and tracheoesophageal speech. RESULTS: Of 349 cases, 193 (55.3%) PE defects were circumferential and 156 (44.7%) were partial. The majority of defects resulted from laryngopharyngectomy (72.5%), most reconstructed with the anterolateral thigh flap (60%). There were 15.5% total esophagectomies, all of which received supercharged jejunal flaps. Of 349 patients, 81 patients (23.2%) had recipient site complications and 51 patients (14.6%) had donor site complications. The fistula rate trended higher in circumferential defects (11 vs. 6%, p = 0.144), and the stricture rate was significantly higher (9.3 vs. 3.8%, p = 0.044). In total, 302 patients (86.5%) had an oral diet after reconstruction, 64 (18%) of whom required supplemental tube feeds. Among 147 patients (42%) who received tracheoesophageal puncture (TEP), 19 (12.9%) eventually failed. Approximately 87% of patients with TEPs achieved fluent speech. The 5-year survival was low for all groups, ranging from 0 to 35%. CONCLUSIONS: PE reconstruction can be performed safely, and most patients will achieve functional speech and swallowing. Swallowing function is worse when the larynx is removed, and the stricture rate is higher with circumferential defects. Specific technical measures can reduce the rate of common complications. LEVEL OF EVIDENCE: The level of evidence of this article was level III.

Bilateral Cleft Lip Repair: Lessons from History.

LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Describe the bilateral cleft lip and nasal deformity and associated anatomical variations. 2. Understand the history and evolution of the bilateral cleft lip repair. 3. Understand the key principles of the surgical repair. SUMMARY: This article describes characteristics of the bilateral cleft lip and nasal deformity and its management, including presurgical orthopedics, operative techniques, and postsurgical care.

Update in Management of Craniosynostosis.

LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Understand the craniofacial dysmorphology of craniosynostosis, and the variation of each type. 2. Identify the functional concerns and learn the rationale behind timing of operative intervention. 3. Approach each dysmorphology critically and identify the operative intervention needed to improve form and function 4. Understand and address the specific issues related to syndromic craniosynostosis and be able to delineate management plan. SUMMARY: Craniosynostosis is a condition in which premature fusion of one or more cranial sutures lead to abnormal head shape and growth restriction of the brain. Nonsyndromic craniosynostosis occurs in isolation, and usually involves a single suture, whereas syndromic craniosynostosis may involve multiple sutures and is associated with extracraniofacial findings. Although surgical management can be similar, the treatment plan must take into consideration issues specific to the syndromes. This article aims to provide a concise overview of the authors' current understanding regarding the presentation, treatment principle, surgical option, and debates in craniosynostosis.

Dual Innervation of Free Functional Muscle Flaps in Facial Paralysis.

Dual innervation in free muscle flap facial reanimation has been used to create a functional synergy between the powerful commissure excursion that can be achieved with the masseter nerve and the spontaneity that can be derived from a cross-face nerve graft. The gracilis has been the most frequently used muscle flap, and multiple combinations of neurorrhaphies have been described, including the masseter to the obturator (end-to-end) combined with a cross-face nerve graft to the obturator (end-to-side) and vice versa. Single and staged approaches have been reported. Minimally, dual innervation is as effective as using the motor nerve to masseter alone.

Update in Unilateral Cleft Lip Surgery.

LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Describe the unilateral cleft lip and nasal deformity and associated anatomical variations. 2. Understand the history and evolution of the unilateral cleft lip repair. 3. List different presurgical treatment options. 4. Differentiate between surgical techniques. SUMMARY: This article describes characteristics of the unilateral cleft lip and nasal deformity and its management, including presurgical orthopedics, operative techniques, and postsurgical care. The rotation-advancement and straight-line repairs are discussed in detail, as are the current concepts in primary cleft nose repair.

Achieving Predictability in Augmentation Mastopexy: An Update.

BACKGROUND: One-stage augmentation mastopexy is a challenging procedure, with the highest cited revision rates in plastic surgery. This is because when mastopexy and augmentation are performed together, they lead to opposing forces, which must be balanced carefully to avoid complications. The goal of this study was to revisit a previously described predictable and safe approach to one-stage augmentation mastopexy, and provide long-term updated results. METHODS: One hundred seventy-one patients who underwent augmentation mastopexy, performed by a single surgeon (R.J.R.), were included in this retrospective review between January 2005 and January 2019. Wise pattern mastopexy with wide pedicle was performed before placement of a small subpectoral implant. Demographic information, preoperative breast measurements, intraoperative technique, implant choice, and postoperative complications were analyzed. Specifically, postoperative measurement of vertical limbs was performed to assess long-term elongation of the lower breast pole. RESULTS: Cumulative complication rate was 11.7%. This rate decreased to 6% in the last 88 patients in this series as the technique matured. The most common complication was revision for implant size exchange. Long-term follow-up demonstrated elongation of nipple-to-inframammary fold distance by 1.0-2.2 cm. There was no recurrence of ptosis requiring reoperation. CONCLUSIONS: This one-stage augmentation mastopexy technique provides a safe and reliable surgical approach with predictable and minimal elongation of the lower breast pole. The reoperation rate of this technique is less than half of >20% revision rate currently cited in the literature.

Facial Fat Augmentation Finesse.

Volume restoration with autologous fat augmentation has become a key technique in facial rejuvenation. It is highly versatile and useful as an independent operation or as an adjunct to rhytidectomy and blepharoplasty. The goal of this article and the accompanying videos is to provide a detailed step-by-step guide to achieving predictable and optimized surgical results.

Dietary Supplements and Current Available Evidence.

Dietary supplements are frequently used in surgical patients. Surgeons should be up to date with regard to the efficacies and potential complications related to these supplements. This article provides the most updated practices and evidence of commonly used supplements.

Applications of Fat Grafting in Pediatric Patients.

Autologous fat grafting has become a widely utilized technique for a variety of cosmetic and reconstructive procedures. Its potential for volume restoration and tissue regeneration has made it a popular method for treating soft tissue defects in both adult and pediatric populations. While autologous fat grafting in the pediatric setting is not as well characterized as it is in the adult setting, various reports have demonstrated the safety and utility of its applications in nonadult patient populations. In this article, we present the first comprehensive review of the current applications of autologous fat grafting in pediatric patients. Specific challenges to fat grafting in the pediatric setting and future applications will also be discussed.

Follow-Up Study: One-Step Salvage of Infected Prosthetic Breast Reconstructions Using Antibiotic-Impregnated Polymethylmethacrylate Plates and Concurrent Tissue Expander Exchange.

BACKGROUND: Periprosthetic infections represent major complications in breast reconstruction, frequently leading to expander-implant loss. No consensus regarding a management algorithm for attempted salvage currently exists. This study assessed outcomes of the authors' salvage protocol using an antibiotic-impregnated polymethylmethacrylate implant with expander device exchange. METHODS: A retrospective chart review identified infected implant-based breast reconstruction cases treated between 2009 and 2017. Of 626 cases initially identified, a total of 62 cases had severe prosthetic infections, and underwent either prosthetic salvage (n = 45) or immediate explantation (n = 17). All the prosthetic salvage patients received intravenous antibiotics followed by surgical débridement, insertion of polymethylmethacrylate plates, device exchange, and postoperative antibiotics. After clinical resolution of infection, tissue expansion was performed, with the polymethylmethacrylate plates remaining in situ until exchanged for permanent implants. RESULTS: The authors' study demonstrated a primary infection clearance rate of 82.2 percent (n = 37). Compared to the traditional explantation group, a significantly higher percentage of the salvage patients completed final reconstruction (84.4 percent versus 35.3 percent; p < 0.001). Fewer patients abandoned reconstruction efforts after infection clearance (2.2 percent versus 58.8 percent; p < 0.001). The majority of cases (78.8 percent) that succeeded the salvage protocol ultimately received implant-based reconstruction; 62.5 percent that failed the salvage protocol still went on to receive autologous tissue reconstruction. CONCLUSIONS: Sustained local antibiotic delivery using polymethylmethacrylate implants and expander device exchange can successfully salvage an infected breast expander/implant. Compared with the traditional explantation approach, more patients complete final reconstruction. Other benefits include preserved skin envelope integrity and possibly improved long-term aesthetic outcomes. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, III.

Is Tissue Expansion Worth It? Comparative Outcomes of Skin-preserving versus Delayed Autologous Breast Reconstruction.

The requirement for postmastectomy radiation therapy (PMRT) at the time of mastectomy is often unknown. Autologous reconstruction is preferred in the setting of radiotherapy by providing healthy vascularized tissue to the chest. To maximize mastectomy skin preservation, tissue expander (TE) placement maintains the breast pocket until definitive reconstruction. This study aims to compare outcomes of skin-preserving delayed versus standard delayed autologous breast reconstruction in the setting of PMRT. METHODS: A retrospective review of a prospective database was performed of two patient cohorts at a single center between 2006 and 2016. Inclusion criteria were locally advanced breast cancer patients who completed PMRT and free autologous reconstruction. Primary outcomes were major intraoperative and postoperative TE and flap complications. RESULTS: Over 10 years, 241 patients underwent mastectomy and PMRT. Standard delayed autologous breast reconstruction was performed in 131 breasts (non-TE group). Skin-preserving delayed autologous reconstruction was performed in 113 breasts (TE group). The TE group was associated with a higher incidence of intraoperative complications during flap reconstruction (P = 0.002) and had a higher venous thrombosis incidence than the non-TE cohort (P = 0.007). Other major postoperative complications were not significantly different between the two groups. TE patients had 7.5 times higher risk of intraoperative complications and an 18.6% TE loss rate. CONCLUSIONS: We identified higher intraoperative flap complications and a high rate of TE loss in patients who underwent skin-preserving delayed autologous breast reconstruction. The benefit of mastectomy skin preservation needs to be weighed against the increased risk of TE loss and higher rates of flap thrombosis.

The Supercharged Pedicled Jejunal Flap for Total Esophageal Reconstruction: A Retrospective Review of 100 Cases.

BACKGROUND: When gastric pull-up is unsuccessful or unsuitable for total esophageal reconstruction, a supercharged pedicled jejunum can be used to reestablish gastrointestinal continuity. The authors reviewed their technique and outcomes of the supercharged pedicled jejunum for total esophageal reconstruction. METHODS: A retrospective review of a prospectively maintained database was performed of 100 patients who underwent supercharged pedicled jejunum for total esophageal reconstruction between 2000 and 2017 at the Texas Medical Center. Patient characteristics, technical details, and outcomes were analyzed. RESULTS: Mean patient age was 59.5 ± 11.4 years. Forty-two patients (42 percent) had surgical complications (18 percent at the recipient site, 13 percent at the donor site, and 11 percent at both). Medical complications occurred in 28 patients (28 percent). A major surgical complication occurred in 20 patients (20 percent). The average length of stay was 15 days (range, 6 to 152 days). At last follow-up, 20 patients (20 percent) had metastatic disease and six (6 percent) had local recurrence. Fifty-four patients (54 percent) died during the follow-up period. Of 79 patients with follow-up longer than 6 months, 68 (86 percent) tolerated a solid or soft oral diet, with a 16 percent tube feed-dependence rate. Overall survival at 1, 3, and 5 years was 78.8, 53.7, and 33.1 percent, respectively. The median survival time was 38.7 months. CONCLUSIONS: The authors present their experience with 100 supercharged pedicled jejunums for total esophageal reconstruction. Functional outcomes are comparable to, or better than, other salvage modalities. With careful multidisciplinary planning and meticulous, well-orchestrated surgical technique, swallowing function can be restored to provide quality of life in patients with few remaining surgical options.

Differences in the Management of Pediatric Facial Trauma.

Craniofacial trauma is common in the pediatric population, with most cases limited to soft tissue and dentoalveolar injury. Although facial fractures are relatively rare in children compared with adults, they are often associated with severe injury and cause significant morbidity and disability. Initial evaluation of a child with facial trauma generally involves stabilizing the patient and identifying any severe concomitant injuries before diagnosing and managing facial injuries. The management of pediatric facial fractures is relatively more conservative than that of adults, and nonsurgical management is preferred when possible to prevent the disruption of future growth and development. Outcomes depend on the site of the injury, management plan, and subsequent growth, so children must be followed longitudinally for monitoring and the identification of any complications.

Multidisciplinary care of craniosynostosis.

The management of craniosynostosis, especially in the setting of craniofacial syndromes, is ideally done in a multidisciplinary clinic with a team focused toward comprehensive care. Craniosynostosis is a congenital disorder of the cranium, caused by the premature fusion of one or more cranial sutures. This fusion results in abnormal cranial growth due to the inability of the involved sutures to accommodate the growing brain. Skull growth occurs only at the patent sutures, resulting in an abnormal head shape. If cranial growth is severely restricted, as seen in multisuture craniosynostosis, elevation in intracranial pressure can occur. Whereas most patients treated in a multidisciplinary craniofacial clinic have non-syndromic or isolated craniosynostosis, the most challenging patients are those with syndromic craniosynostosis. The purpose of this article was to discuss the multidisciplinary team care required to treat both syndromic and non-syndromic craniosynostosis.

Optimizing Outcomes in Pharyngoesophageal Reconstruction and Neck Resurfacing: 10-Year Experience of 294 Cases.

BACKGROUND: Pharyngoesophageal reconstruction is a complex endeavor that poses many challenges. This 10-year series examines factors impacting the need for neck skin resurfacing and evaluates the impact of reconstructive modalities on outcomes. METHODS: A review identified 294 patients who underwent pharyngoesophageal reconstruction from 2002 to 2012. Patients were divided based on neck skin resurfacing requirements. Patients undergoing neck resurfacing were further subdivided into reconstructive technique, including a second skin paddle or muscle component from the same free flap pedicle, a local flap, or a second free flap. All groups were compared by comorbidities, complications, and functional outcomes. RESULTS: Of 294 patients, 179 (60.9 percent) required neck skin resurfacing. In the resurfaced group, there were 90 circumferential defects (50.3 percent) and 89 partial defects (49.7 percent). In the resurfaced group, 110 (61.4 percent) underwent reconstruction with a second skin paddle from the same free flap pedicle, 21 (11.7 percent) underwent reconstruction with a muscle component from the same pedicle, and 25 (13.9 percent) received a pectoralis major flap. There were five external paddle flap losses in the resurfaced group (2.8 percent) and no internal flap losses. Overall complications were similar among groups. The resurfaced group had a lower pharyngocutaneous fistula rate (4.5 percent) compared with the primary closure group (11.3 percent) (p = 0.026). Prior neck surgery and radiation therapy were strong predictors of neck skin resurfacing (p < 0.001). CONCLUSIONS: Neck resurfacing is often required in salvage pharyngoesophageal reconstruction. Providing additional vascularized tissue over the neoconduit is predictive of lower pharyngocutaneous fistula rates. An algorithmic approach to neck resurfacing is presented.