Derivation of two human induced pluripotent stem cell lines carrying a missense mutation in FHL1 (c.377G > A, p.C126Y) linked to familial muscular dystrophy.

FHL1 gene locates in the Xq26 region and encodes for four and half LIM domain protein 1. It plays a crucial role in muscle cells and mutations in FHL1 are related to muscular dystrophy (MD). Peripheral blood mononuclear cells (PBMCs) were obtained from 2 family patients with MD that carry a pathogenic missense mutation in FHL1 (c.377G > A, p.C126Y). Induced pluripotent stem cells (iPSCs) were generated by PBMCs reprogramming using the lentiviral-hSTEMCCA-loxP vector, obtaining FHL1-T and FHL1-V iPSCs lines from patients. FHL1 genotype was maintained, and stemness and pluripotency were confirmed in both iPSCs lines.

Caustics and wavefronts produced by multiple refraction-reflections through plane-parallel plates.

We study the formation of caustics and wavefronts produced by multiple refraction-reflections through a plane-parallel transparent plate, assuming a point source placed at an arbitrary position along the optical axis. The caustic surfaces are obtained by using the envelope's method. Subsequently, the wavefronts are directly related to the involutes, which are associated with the envelopes for all the rays. Hence by using the Malus-Dupin theorem, we obtain their respective wavefronts produced by multiple refraction-reflections through a plane-parallel transparent plate. On the other hand, we implement Huygens' principle to obtain the wavefronts leaving the plate after undergoing multiple reflections inside the plate, which we have called zero-distance phase wavefronts. Finally, we establish the correspondence between the wavefronts obtained by Huygens' principle and the involutes associated with caustic surfaces; they are brought in coincidence assuming parallel curves from each other.

Isokinetic Training Program to Improve the Physical Function and Muscular Performance of an Individual with Partial Injury of the Medial Meniscus: A Case Report.

Background and Purpose: One of the best alternatives for the treatment of meniscal injuries in relation to surgical procedures, is exercise. This case report aimed to describe the effects of isokinetic training and a neuromuscular/proprioceptive exercise program on muscle performance and physical function in an individual after a rupture of the posterior horn of the medial meniscus. Study Design: Case report. Case Description: A 40-year-old man injured his right knee during a soccer match, with a partial lesion of the medial meniscus confirmed by magnetic resonance imaging. He completed an isokinetic training program in addition to exercises that targeted proprioception (22 sessions, 11 weeks) to improve physical function and performance, which were assessed before and after treatment and at a six-month follow-up. An individual with similar anthropometric characteristics was chosen to be used as a control for understanding the patient's assessment values. Outcomes: Muscular performance of the knee flexors and extensors was evaluated isokinetically using the Biodex System-4 in a concentric mode at angular velocities of 60, 120, and 300 °/s . The main results indicated that after 11 weeks, the peak torque normalized to body mass (PT/BM), at 60 °/s of the knee extensors remained unchanged (2.54 N.m/kg) (below the control value - 3.06 N.m/kg), and at the six-month follow-up, increased by approximately 20% (3.08 N.m/kg). For the hamstrings, at 60 °/s, an increase of 18 % occurred after intervention (1.98 N.m/kg) and by approximately 30 % at the six-month follow-up (2.12 N.m/kg) - values much higher than the control 1.55 N.m/kg). This increase in the PT/BM was also reflected in the Hamstrings:Quadriceps ratio (78 %) after treatment which improved at follow-up (68 %). Discussion: The results showed that the isokinetic training and neuromuscular/proprioceptive exercises improved the muscle performance of the knee flexors and extensors, after eleven weeks of intervention, and remained (or continued to improve) at the six-month follow-up. Level of evidence: 5, single case report.

Generation of two edited iPSCs lines by CRISPR/Cas9 with point mutations in PKP2 gene for arrhythmogenic cardiomyopathy in vitro modeling.

The arrhythmogenic cardiomyopathy (ACM) is an inherited heart muscle disease characterized by the progressive replacement of contractile myocardium by fibro-fatty adipose tissue, that generates ventricular arrhythmias and sudden death in patients. The ACM has a genetic origin with alterations in desmosomal genes with the most commonly mutated being the PKP2 gene. We generated two CRISPR/Cas9 edited iPSCs lines, one iPSC line with a point mutation in PKP2 reported in patients with ACM and another iPSC line with a premature stop codon to knock-out the same gene.

EFFECTS OF BACTERIAL LIPOPOLYSACCHARIDE AND SHIGA TOXIN ON INDUCED PLURIPOTENT STEM CELL-DERIVED MESENCHYMAL STEM CELLS.

ABSTRACT: Background : Mesenchymal stem cells (MSCs) can be activated by different bacterial toxins. Lipopolysaccharides and Shiga Toxin (Stx) are the main toxins necessary for hemolytic uremic syndrome development. The main etiological event in this disease is endothelial damage that causes glomerular destruction. Considering the repairing properties of MSC, we aimed to study the response of MSC derived from induced pluripotent stem cells (iPSC-MSC) to LPS and/or Stx and its effect on the restoration of injured endothelial cells. Methods : iPSC-MSC were treated with LPS and or/Stx for 24 h and secretion of cytokines, adhesion, and migration were measured in response to these toxins. In addition, conditioned media from treated iPSC-MSC were collected and used for proteomics analysis and evaluation of endothelial cell healing and tubulogenesis using human microvascular endothelial cells 1 as a source of endothelial cells. Results : The results obtained showed that LPS induced a proinflammatory profile on iPSC-MSC, whereas Stx effects were less evident, even though cells expressed the Gb 3 receptor. Moreover, LPS induced on iPSC-MSC an increment in migration and adhesion to a gelatin substrate. Addition of conditioned media of iPSC-MSC treated with LPS + Stx, decreased the capacity of human microvascular endothelial cells 1 to close a wound, and did not favor tubulogenesis. Proteomic analysis of iPSC-MSC treated with LPS and/or Stx revealed specific protein secretion patterns that support the functional results described. Conclusions : iPSC-MSC activated by LPS acquired a proinflammatory profile that induces migration and adhesion to extracellular matrix proteins but the addition of Stx did not activate any repair program to ameliorate endothelial damage, indicating that the use of iPSC-MSC to regenerate endothelial injury caused by LPS and/or Stx in hemolytic uremic syndrome could not be the best option to consider to regenerate a tissue injury.

Special Considerations in Face Transplantation: A Systematic Review.

ABSTRACT: Vascularized composite allotransplantation of the face is an exceedingly complex procedure, requiring extensive planning and surgical precision in order to successfully manage patients with facial disfigurements. This review aims to present an overview of the salient anatomic considerations in facial transplantation, as well as give attention to unique patient populations and special considerations.A literature review was performed in search of articles pertaining to considerations in facial transplantation using the databases PubMed, Web of Science, and Cochrane. Articles selected for further review included full-text articles with an emphasis on specific anatomic defects and how they were addressed in the transplant process, as well as management of special patient populations undergoing facial transplantation. In total, 19 articles were deemed appropriate for inclusion.The use of computer-assisted technologies for the planning portion of the procedure, as well as intraoperative efficiency, has yielded favorable results and can be considered as part of the operative plan. The ultimate outcome is dependent upon the synchronization of subunits of the allograft and the desired functional outcomes, including osseous, ocular, oral, and otologic considerations. Management of specific pathology and subgroups of patients are critical aspects. Although pediatric face transplantation has not yet been performed, it is a likely a future step in the evolution of this procedure.When performing a face transplantation, many components must be considered pre-, intra-, and post-operatively. This systematic review presents specific anatomic considerations, as well as information about special patient populations within this crosssection of multidisciplinary microsurgery, psychiatry, and transplant immunology.

Cranioplasty Outcomes From 500 Consecutive Neuroplastic Surgery Patients.

BACKGROUND: Cranioplasty is critical to cerebral protection and restoring intracranial physiology, yet this procedure is fraught with a high risk of complications. The field of neuroplastic surgery was created to improve skull and scalp reconstruction outcomes in adult neurosurgical patients, with the hypothesis that a multidisciplinary team approach could help decrease complications. OBJECTIVE: To determine outcomes from a cohort of cranioplasty surgeries performed by a neuroplastic surgery team using a consistent surgical technique and approach. METHODS: The authors reviewed 500 consecutive adult neuroplastic surgery cranioplasties that were performed between January 2012 and September 2020. Data were abstracted from a prospectively maintained database. Univariate analysis was performed to determine association between demographic, medical, and surgical factors and odds of revision surgery. RESULTS: Patients were followed for an average of 24 months. Overall, there was a reoperation rate of 15.2% (n = 76), with the most frequent complications being infection (7.8%, n = 39), epidural hematoma (2.2%, n = 11), and wound dehiscence (1.8%, n = 9). New onset seizures occurred in 6 (1.2%) patients.Several variables were associated with increased odds of revision surgery, including lower body mass ratio, 2 or more cranial surgeries, presence of hydrocephalus shunts, scalp tissue defects, large-sized skull defect, and autologous bone flaps. importantly, implants with embedded neurotechnology were not associated with increased odds of reoperation. CONCLUSIONS: These results allow for comparison of multiple factors that impact risk of complications after cranioplasty and lay the foundation for development of a cranioplasty risk stratification scheme. Further research in neuroplastic surgery is warranted to examine how designated centers concentrating on adult neuro-cranial reconstruction and multidisciplinary collaboration may lead to improved cranioplasty outcomes and decreased risks of complications in neurosurgical patients.

Approach for designing thick cemented doublet lenses based on the caustic surface.

A new, to the best of our knowledge, method for designing a thick-lens achromatic doublet based on the concept of a caustic surface to correct both third- and fifth-order spherical aberration is presented. We consider two different wavelengths brought into coincidence at the back focal length instead of the effective focal length as it is usually done, to calculate the radii of curvature assuming predefined values for axial thicknesses and their indices of refraction for both lenses. Alternatively, we apply Taylor's series around the optical axis, and to vanish the approximate caustic surface, we obtain the values for the conic constants, which reduce at third- and fifth-order spherical aberration. Two designs for cemented doublets are provided assuming that the lenses are cemented. Finally, we propose a method to qualitatively test doublet lenses by using null screens, considering to place the detection plane at arbitrary positions.

Ronchi-Hartmann type null screens for testing a plano-freeform surface with a detection plane inside a caustic surface.

We have implemented an exact ray trace through a plano-freeform surface for an incident plane wavefront. We obtain two caustic surfaces and provide the critical points related to the ray tracing process. Additionally, we study the propagation of the refracted wavefronts through the plane-curved surface. Finally, by using the Ronchi-Hartmann type null screen and placing the detection plane within the caustic region, we have evaluated the shape of a plano-freeform optical surface under test, obtaining an RMS difference in sagitta value of 6.3 µm.

celldeath: A tool for detection of cell death in transmitted light microscopy images by deep learning-based visual recognition.

Cell death experiments are routinely done in many labs around the world, these experiments are the backbone of many assays for drug development. Cell death detection is usually performed in many ways, and requires time and reagents. However, cell death is preceded by slight morphological changes in cell shape and texture. In this paper, we trained a neural network to classify cells undergoing cell death. We found that the network was able to highly predict cell death after one hour of exposure to camptothecin. Moreover, this prediction largely outperforms human ability. Finally, we provide a simple python tool that can broadly be used to detect cell death.

Airway Management of COVID-19 Positive Patients Aboard USNS COMFORT Mission to New York City.

During the USNS Comfort mobilization to New York City in response to the pandemic, the ship's medical team cared for over 35 mechanically ventilated corona virus disease 2019 (COVID-19) infected patients. Due to the better than expected mortality rates of these patients, tracheotomy for prolonged intubation or other indicated interventional bronchoscopies were performed on 7 COVID positive patients, as well as 2 with negative screening tests. No member of the health care team subsequently became symptomatic or tested positive for COVID-19. This was in part due to the formation of a dedicated surgical airway team, use of standardized procedural techniques and personal protective equipment (PPE), and construction of a negative pressure operating room within the COVID-19 isolation ward on the ship. This experience shows that tracheotomies and other aerosolizing procedures can be performed with due concern for patient and provider safety, regardless of patient's COVID status.

Neuroplastic Surgery Aboard USNS Comfort During the COVID-19 Pandemic in New York City.

In December of 2019, a novel virus known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) produced a cluster of viral pneumonia cases in Wuhan, China. In the United States (US), New York City was particularly devastated, with the volume and acuity of patients placing an unprecedented strain on the hospital system and health care workers. In response to this crisis, USNS Comfort (T-AH 20) was deployed to New York City with a 1100 member medical team to augment local hospitals. Comfort's mission to New York City was dynamic, and required special adaptation to care for both COVID positive and COVID negative patients. Neuroplastic surgery procedures were indicated in both COVID positive and COVID negative patients, and lessons learned with regard to performance of complex surgery in an unfamiliar environment consisted of developing a thorough understanding of ones capabilities, and working with a highly skilled team of Navy surgeons, anesthesiologists, and surgical support staff, in order to provide high quality care in a deployment platform.

Misconceptions About Negative Pressure Rooms and Their Impact Aboard USNS Comfort in New York City.

The outbreak of novel coronavirus disease 2019 (COVID-19) has had a momentous impact on the field of otolaryngology due to the high number of aerosol-generating procedures involving the upper aerodigestive tract. These procedures bear significant risk to the provider and clinical environment due to the possibility of viral aerosolization. While significant attention has been appropriately paid to personal protective equipment during this pandemic, an understanding of industrial hygiene is also necessary for the safe delivery of health care to mitigate the risk of exposure to other patients and health care workers. We provide a review of air ventilation practices and their role in reducing pathogen spread. In addition, we share our experiences with effectively treating COVID-19-positive patients aboard the USNS Comfort through proper environment control measures.

PIWI-interacting RNAs are differentially expressed during cardiac differentiation of human pluripotent stem cells.

PIWI-interacting RNAs (piRNAs) are a class of non-coding RNAs initially thought to be restricted exclusively to germline cells. In recent years, accumulating evidence has demonstrated that piRNAs are actually expressed in pluripotent, neural, cardiac and even cancer cells. However, controversy remains around the existence and function of somatic piRNAs. Using small RNA-seq samples from H9 pluripotent cells differentiated to mesoderm progenitors and cardiomyocytes we identified the expression of 447 piRNA transcripts, of which 241 were detected in pluripotency, 218 in mesoderm and 171 in cardiac cells. The majority of them originated from the sense strand of protein coding and lncRNAs genes in all stages of differentiation, though no evidences of amplification loop (ping-pong) were found. Genes hosting piRNA transcripts in cardiac samples were related to critical biological processes in the heart, like contraction and cardiac muscle development. Our results indicate that these piRNAs might have a role in fine-tuning the expression of genes involved in differentiation of pluripotent cells to cardiomyocytes.

Risk of Complications in Primary Versus Revision-Type Cranioplasty.

INTRODUCTION: Cranioplasty (CP) is a multifaceted procedure in a heterogenous patient population, with a high risk for complication. However, no previous large-scale studies have compared outcomes in primary (ie, first attempt) CP versus revision CP (ie, following previous attempts). The authors, therefore, analyzed long-term outcomes of 506 consecutive primary and revision CPs, performed by a single surgeon. METHODS: All CPs performed between 2012 and 2019 were analyzed under IRB protocol approval. Surgeries were categorized as either primary (no previous CP; n = 279) or revision CP (at least one previous CP; n = 227). Complications were defined as either major or minor. Subgroup analyses investigated whether or not CP complication risk directly correlated with the number of previous neuro-cranial surgeries and/or CP attempts. RESULTS: The primary CP group experienced a major complication rate of 9% (26/279). In comparison, the revision CP group demonstrated a major complication rate of 32% (73/227). For the revision CP group, the rate of major complications rose with each additional surgery, from 4% (1 prior surgery) to 17% (2 prior surgeries) to 39% (3-4 prior surgeries) to 47% (≥5 prior surgeries). CONCLUSION: In a review of 506 consecutive cases, patients undergoing revision CP had a 3-fold increase in incidence of major complications, as compared to those undergoing primary CP. These results provide critical insight into overall CP risk stratification and may guide preoperative risk-benefit discussions. Furthermore, these findings may support a center-of-excellence care model, particularly for those patients with a history of previous neuro-cranial surgeries and/or CP attempts.

Sagittal and tangential foci produced by tilted plane wavefronts refracted through simple lenses.

We study the formation of caustic and wavefront surfaces produced by a tilted plane wavefront propagating through spherical positive lenses. The shape of the caustic surface is a function of the indices of refraction, the geometrical parameters of the lens involved in the process of refraction, and the obliquity angle with respect to the optical axis, as we expect. We provide exact and approximate analytic equations for tangential and sagittal focal surfaces and also for Petzval field curvature considering arbitrary lenses.

Adult Cranioplasty.

OBJECTIVES: After reading this article, the participant should be able to: Understand the etiology of cranial defects. Understand the anatomy of the cranium. Understand the importance of the preoperative workup in the cranial reconstruction decision-making process. Describe the options available for calvarial reconstruction including autologous and alloplastic materials. Describe the basic differences between available alloplastic materials. Understand the intraoperative and postoperative complications that may arise during cranioplasty. SUMMARY: Cranial defects can arise from a variety of causes, yielding a diverse group of patients who require cranioplasty. The goals of calvarial reconstruction are to protect the underlying brain, to restore the aesthetic contour of the calvarium, and/or to treat postcraniectomy cerebrospinal fluid circulation abnormalities that may be symptomatic. Options for calvarial reconstruction include the autogenous bone flap that was removed for access, autologous bone grafting, and a variety of alloplastic materials such as titanium, hydroxyapatite, polymethylmethacrylate, polyether ether ketone, and high-density porous polyethylene. A detailed preoperative workup and discussion with the patient is important to choosing the appropriate reconstructive path.

LMO2 Confers Synthetic Lethality to PARP Inhibition in DLBCL.

Deficiency in DNA double-strand break (DSB) repair mechanisms has been widely exploited for the treatment of different malignances, including homologous recombination (HR)-deficient breast and ovarian cancers. Here we demonstrate that diffuse large B cell lymphomas (DLBCLs) expressing LMO2 protein are functionally deficient in HR-mediated DSB repair. Mechanistically, LMO2 inhibits BRCA1 recruitment to DSBs by interacting with 53BP1 during repair. Similar to BRCA1-deficient cells, LMO2-positive DLBCLs and T cell acute lymphoblastic leukemia (T-ALL) cells exhibit a high sensitivity to poly(ADP-ribose) polymerase (PARP) inhibitors. Furthermore, chemotherapy and PARP inhibitors synergize to inhibit the growth of LMO2-positive tumors. Together, our results reveal that LMO2 expression predicts HR deficiency and the potential therapeutic use of PARP inhibitors in DLBCL and T-ALL.

First-in-Human Experience With Integration of a Hydrocephalus Shunt Device Within a Customized Cranial Implant.

BACKGROUND: Implantable shunt devices are critical and life saving for hydrocephalus patients. However, these devices are fraught with high complication rates including scalp dehiscence, exposure, and extrusion. In fact, high shunt valve profiles are correlated with increased complications compared to those with lower profiles. As such, we sought a new method for integrating shunt valves for those challenging patients presenting with scalp-related complications. OBJECTIVE: To safely implant and integrate a hydrocephalus shunt valve device within a customized cranial implant, in an effort to limit its high-profile nature as a main contributor to shunt failure and scalp breakdown, and at the same time, improve patient satisfaction by preventing contour deformity. METHODS: A 64-yr-old male presented with an extruding hydrocephalus shunt valve and chronic, open scalp wound. The shunt valve was removed and temporary shunt externalization was performed. He received 2 wk of culture-directed antibiotics. Next, a contralateral craniectomy was performed allowing a new shunt valve system to be implanted within a low-profile, customized cranial implant. All efforts were made, at the patient's request, to decrease the high-profile nature of the shunt valve contributing to his most recent complication. RESULTS: First-in-human implantation was performed without complication. Postoperative shunt identification and programming was uncomplicated. The high-profile nature of the shunt valve was decreased by 87%. At 10 mo, the patient has experienced no complications and is extremely satisfied with his appearance. CONCLUSION: This first-in-human experience suggests that a high-profile hydrocephalus shunt device may be safely integrated within a customized cranial implant.