Current Protocols: Alopecia Areata Mouse Models for Drug Efficacy and Mechanism Studies.

Alopecia areata is the second most common form of hair loss in humans after androgenetic alopecia. Although a variety of animal models for alopecia areata have been described, currently the C3H/HeJ mouse model is the most commonly used and accepted. Spontaneous hair loss occurs in 15%-25% of older mice in which the lesions wax and wane, similar to the human disease, with alopecia being more common and severe in female mice. Full-thickness skin grafts from mice with spontaneous alopecia areata to young, normal-haired, histocompatible mice provide a highly reproducible model with progressive lesions that makes it useful for drug efficacy and mechanism-based studies. As alopecia areata is a cell-mediated autoimmune disease, transfer of cultured lymph node cells from affected mice to unaffected, histocompatible recipients also promotes disease development and provides an alternative, nonsurgical protocol. Protocols are presented to produce these models such that they can be used to study alopecia areata and to develop novel drug therapies. © 2024 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Full-thickness skin grafts to reproducibly induce alopecia areata in C3H/HeJ mice Basic Protocol 2: Adoptive transfer of cultured lymphoid cells provides a nonsurgical method to induce alopecia areata in C3H/HeJ mice.

Low Threshold for Laparoscopic Exploration in Dementia Patients With Unexplained Abdominal Pain and Risk Factors for Acute Mesenteric Ischemia.

Acute mesenteric ischemia (AMI) is associated with a high mortality and morbidity. There are limited studies on the presentation and management of elderly dementia patients with AMI. The following case of an 88-year-old female with dementia presenting with AMI highlights the challenges in the care of elderly dementia patients with AMI, the importance of identifying risk factors and hallmarks of acute mesenteric ischemia early in the clinical course, and suggests that aggressive workup with diagnostic laparoscopy is crucial to timely diagnosis and effective care.

Financial Toxicity in Emergency General Surgery: Novel Propensity-Matched Outcome Comparison.

BACKGROUND: Financial toxicity describes the harmful effect of individual treatment costs and fiscal burdens that have a compounding negative impact on outcomes in surgery. While this phenomenon has been widely studied in surgical oncology, the purpose of this study was to perform a novel exploration of the impact of financial toxicity in emergency general surgery (EGS) patients throughout the US. STUDY DESIGN: The Nationwide Readmissions Database for January and February 2018 was queried for all EGS patients aged 18 to 65 years. One-to-one propensity matching was performed with and without risk for financial toxicity. The primary outcome was mortality, and the secondary outcomes were venous thromboembolism (VTE), prolonged length of stay (LOS), and readmission within 30 days. RESULTS: There were 24,154 EGS patients propensity matched. The mortality rate was 0.2% (n = 39), and the rate of VTE was 0.5% (n = 113). With financial toxicity, there was no statistically significant difference for mortality (p = 0.08) or VTE (p = 0.30). The rate of prolonged LOS was 6.2% (n = 824), and the risk was increased with financial toxicity (risk ratio 1.24 [1.12 to 1.37]; p < 0.001). The readmission rate was 7.0% (n = 926), and the risk with financial toxicity was increased (risk ratio 1.21 [1.10 to 1.33]; p < 0.001). The mean count of comorbidities per patient per admission during readmission within 1 year with financial toxicity was 2.1 ± 1.9 versus 1.8 ± 1.7 without (p < 0.001). CONCLUSIONS: Despite little difference in the rate of mortality or VTE, EGS patients at risk for financial toxicity have an increased risk of readmission and longer LOS. Fewer comorbidities were identified at index admission than during readmission in patients at risk for financial toxicity. Future studies aimed at reducing this compounding effect of financial toxicity and identifying missed comorbidities have the potential to improve EGS outcomes.

Primary cicatricial alopecias are characterized by dysregulation of shared gene expression pathways.

The primary forms of cicatricial (scarring) alopecia (PCA) are a group of inflammatory, irreversible hair loss disorders characterized by immune cell infiltrates targeting hair follicles (HFs). Lichen planopilaris (LPP), frontal fibrosing alopecia (FFA), and centrifugal cicatricial alopecia (CCCA) are among the main subtypes of PCAs. The pathogenesis of the different types of PCAs are poorly understood, and current treatment regimens yield inconsistent and unsatisfactory results. We performed high-throughput RNA-sequencing on scalp biopsies of a large cohort PCA patients to develop gene expression-based signatures, trained into machine-learning-based predictive models and pathways associated with dysregulated gene expression. We performed morphological and cytokine analysis to define the immune cell populations found in PCA subtypes. We identified a common PCA gene signature that was shared between LPP, FFA, and CCCA, which revealed a significant over-representation of mast cell (MC) genes, as well as downregulation of cholesterogenic pathways and upregulation of fibrosis and immune signaling genes. Immunohistological analyses revealed an increased presence of MCs in PCAs lesions. Our gene expression analyses revealed common pathways associated with PCAs, with a strong association with MCs. The indistinguishable differences in gene expression profiles and immune cell signatures between LPP, FFA, and CCCA suggest that similar treatment regimens may be effective in treating these irreversible forms of hair loss.

Jejuno-jejunal intussusception secondary to recurrent/metastatic melanoma.

Intussusception is uncommon in adults, and usually secondary to a neoplasm. Small bowel obstruction (SBO) is prevalent in adults but rarely due to intussusception. A 55-year-old woman with history of melanoma (four years in remission) presented with abdominal pain and melena. Upper and lower endoscopy was normal. She developed bilious emesis and worsening pain, so she presented to the emergency department. CT abdomen/pelvis identified a SBO with transition point at a small bowel intussusception; she was taken to the operating room. A mid-jejunal intussusception was reduced revealing a mass; resection and primary anastomosis was performed. Final pathology demonstrated a 5.5 cm melanoma, likely metastatic. She required no additional therapy and remains in remission eighteen months later. Intussusception due to metastatic melanoma is rare but should be on the differential for patients with SBO and history of melanoma. Knowledge of this history should prompt consideration for oncologic resection to optimize outcome.

Cerebral Fat Embolism via a Patent Foramen Ovale.

Fat embolism syndrome (FES) is a multisystem process predominantly secondary to long bone/pelvic fractures and orthopedic procedures. A 19-year-old man presents after motor vehicle collision with trace right pneumothorax, right grade 3 kidney laceration, left pubic rami, and right femoral shaft fractures. Right femur closed reduction ensued and he underwent intramedullary nailing; his other injuries were managed nonoperatively. Upon awakening in recovery, he was newly aphasic. Despite negative repeat CT brain, he continued to worsen and became tachycardic and hypoxemic. MRI/MRA brain demonstrated innumerable bilateral frontal, parietal, and occipital acute ischemic infarcts in a starfield pattern. Echocardiogram revealed a PFO. With supportive care, he improved and was discharged with planned outpatient PFO closure. One month later, he had complete symptom resolution with return to neurologic baseline. FES is a potentially devastating condition which may include cerebral fat embolism (CFE) with outcomes varying widely from mortality to complete recovery.

Marine litter on sandy beaches with different human uses and waste management along the Gulf of Nicoya, Costa Rica.

This study was a baseline with quantitative data of marine litter along the Gulf of Nicoya, Costa Rica. The objective of the study was to quantify marine litter and its association with human activities in this estuarine gulf. A total of fourteen sandy beaches were cataloged by the degree of urbanization, tourism intensity, beach cleaning programs, and tributary rivers as possible drivers of marine litter presence. The items of the marine litter were separated and weighted by type. Analysis by the clean coastal index (CCI) and multivariate statistics were applied to find spatial patterns in marine litter in the gulf. On beaches with the highest touristic activity, cigarette butts and straws were the main components. Locations with river plume influence, less frequent cleanup, or waste cans showed more bottles, plastic parts, and sanitary waste than beaches in other conditions. A beach in a fisherman town had recently utilized plastic bags, household goods, and boat parts in the marine litter. A wildlife refuge beach showed only small plastic and coffee foam cup fragments that came with currents from other points in the estuary. River basin management, solid waste disposal programs, and environmental education to avoid single-use items combined with correct waste disposal are needed to reduce marine litter in tropical countries focused on ecological tourism.

Effect of short-course exercise training on the frequency of exacerbations and physical activity in patients with COPD: A randomized controlled trial.

BACKGROUND AND OBJECTIVE: Previous studies have suggested that early pulmonary rehabilitation (PR) programmes post-AECOPD are an effective and safe intervention for reducing hospital admissions and improving quality of life. This study assessed whether a short course of exercise training post-AECOPD with periodic reinforcement exercise training and phone call reminders reduces readmissions and increases physical activity in COPD patients. METHODS: Subjects were randomized into either the (i) intervention group (IG), consisting of 4-8 weeks of training supervised by a physiotherapist and phone contact every 2 weeks by a case manager providing support and reinforcement of continuous exercise at home or (ii) usual care group (UG), which had no input by a physiotherapist or case manager. Readmissions were assessed at 12 months. Activities of all patients were assessed by an activity monitor at baseline, 3 and 12 months. RESULTS: Altogether, 136 subjects were included and randomized (68 in IG and 68 in UG). The age, gender and FEV1 % predicted were 75.0 ± 6.7 years, 132 males and 47.0 ± 16.2%, respectively. The mean number of readmissions for AECOPD (1.06 vs 1.72 times, P = 0.014) was less and time to first readmission was increased (146.8 vs 122.4 days, P = 0.005) in the IG versus UG at 12 months. At 12 months, there was no change in activity measured by activity monitor between the two groups. CONCLUSION: This programme decreased exacerbation frequency and increased the time of readmissions for AECOPD. It did not improve physical activities and exercise tolerance at 12 months.

Nonsurgical Induction of Alopecia Areata in C3H/HeJ Mice via Adoptive Transfer of Cultured Lymphoid Cells.

Surgical induction of alopecia areata (AA) via full-thickness grafting of spontaneous AA-affected C3H/HeJ mouse skin to naïve recipients has been a primary method of transferring the AA disease model phenotype. However, this method is associated with the need to perform an invasive procedure that could negatively impact animal wellbeing. Therefore, a rodent model that rapidly develops AA at a predictable rate and without the need to perform invasive surgical procedures on the mice is essential for studying the pathogenesis of AA. Here we describe a cell injection technique using cultured skin-draining lymph node cells (LNCs) injected intradermally into naïve recipients to induce rapid AA development. The cultured LNCs can reach ~ten fold expansion after 6 days with specific cytokine stimulation. The LNCs derived from a single AA affected mouse donor can induce AA development in more than 80 naïve mice within 2-18 weeks. For comparative control studies, mice receiving cultured LNCs from normal donors remain normally haired. The method enables the production of large numbers of AA mice for use in research and treatment development studies while avoiding the use of surgical procedures. We anticipate that the protocol can also be adapted for use in other mouse autoimmune disease models.

Decitabine mildly attenuates MLL-rearranged acute lymphoblastic leukemia in vivo, and represents a poor chemo-sensitizer.

MLL-rearranged acute lymphoblastic leukemia (ALL) represents a highly aggressive ALL subtype, characterized by aberrant DNA methylation patterns. DNA methyltransferase inhibitors, such as decitabine have previously been demonstrated to be effective in eradicating MLL-rearranged ALL cells in vitro. Here, we assessed the in vivo anti-leukemic potential of low-dose DNA methyltransferase inhibitor decitabine using a xenograft mouse model of human MLL-rearranged ALL. Furthermore, we explored whether prolonged exposure to low-dose decitabine could chemo-sensitize MLL-rearranged ALL cells toward conventional chemotherapy as well as other known epigenetic-based and anti-neoplastic compounds. Our data reveal that decitabine prolonged survival in xenograft mice of MLL-rearranged ALL by 8.5 days (P = .0181), but eventually was insufficient to prevent leukemia out-growth, based on the examination of the MLLAF4 cell line SEM. Furthermore, we observe that prolonged pretreatment of low-dose decitabine mildly sensitized toward the conventional drugs prednisolone, vincristine, daunorubicin, asparaginase, and cytarabine in a panel of MLL-rearranged cell lines. Additionally, we assessed synergistic effects of decitabine with other epigenetic-based or anticancer drugs using high-throughput drug library screens. Validation of the top hits, including histone deacetylase inhibitor panobinostat, BCL2 inhibitor Venetoclax, MEK inhibitor pimasertib, and receptor tyrosine kinase foretinib, revealed additive and moderate synergistic effects for the combination of each drug together with decitabine in a cell line-dependent manner.

Optimization of uranium and plutonium separations using TEVA and UTEVA cartridges for MC-ICP-MS analysis of environmental swipe samples.

The analysis of environmental swipe samples for ultra-trace uranium (U) and plutonium (Pu) determinations is essential in the nuclear safeguards community. While mass spectrometry techniques for U and Pu detection continually improve, established separation methods are seldom reevaluated. Currently, actinide separations within the forensics community predominantly employ either Eichrom TEVA® or UTEVA® resins. The direct optimization of U and Pu separations utilizing both resins has not been widely reported. Here, several methods were explored with goals of increasing analyte recovery, acquiring cleaner blanks, and improving the separation efficiency of ultra-trace levels of U and Pu from environmental swipe samples. The optimized separation methodology of U and Pu was examined using certified reference materials and archived environmental swipe samples.

Evaluation and Specifications for In-Line Uranium Separations Using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) Detection for Trace Elemental Analysis.

Automated introduction platforms integrated with inductively coupled plasma optical emission spectroscopy (ICP-OES) systems are continuously being improved. Expanding on the introduction systems, a newly developed automated ion chromatography system was explored for performing rapid in-line separations coupled to ICP-OES for the detection of trace elements in uranium. Trace elements are separated from a uranium material and the analytes are directed into the ICP-OES for subsequent detection. Detection parameters such as exposure time frequency, wavelength selection, and settling times were explored to gain insight on optimal detection schemes for in-line trace elemental analysis. The methodology was applied in the analysis of a uranium oxide (U3O8) certified reference material, CRM-124. It was found here that the sensitivity and uncertainty of the technique are greatly affected by how the ICP-OES is employed to collect data. Overall it was determined that faster exposure replicates can provide greater peak resolution with higher fidelity measurements but are limited with respect to the total analysis time (i.e., limited in detection timely separations). Zeta scores, which combine accuracy and uncertainty of certified values and experimental values, were used to validate the ICP-OES modes of operation.

Investigating Clinical and Cost Burdens of Law Enforcement-Related K9 Injuries: The Impact of "the Bite" on a Community Hospital.

The decision to introduce canines (K9s) to a law enforcement (LE) agency does not typically involve the evaluation of the fiscal or clinical impact on local hospitals. This study compared injury, cost, and care associated with K9s to a common nonlethal force method, the Thomas A Swift Electrical Rifle (TASER), to highlight the cost and resources required to treat both patient types. Patients treated for LE-related K9 and TASER injuries at a Level I community-based trauma center (2011-2016) were evaluated for level of care required (e.g., surgeon/specialist), clinical interventions, proxy medical costs, and length of stay (LOS). Nearly one-third of K9 patients required tertiary-level medical care. The cost of treating the K9-inflicted injuries (n = 75) was almost twice as costly as care for patients subdued with a TASER (n = 80); the K9 patients had significantly (one-tailed t tests) higher medical costs (P = 0.036), required more medical procedures (P = 0.014), and had longer LOS (P = 0.0046) than the TASER patients. Patients with K9 injuries had higher acuity and were significantly more expensive to treat with longer LOS than TASER injuries. LE agencies considering establishing and operating a K9 unit should initiate discussions with their local medical first responders and health-care facilities regarding the capabilities to treat severe K9 injuries to ensure adequate resource allocation.

Necrotizing Fasciitis Resulting from an Anastomotic Leak after Colorectal Resection.

One of the most feared complications in colorectal surgery is an anastomotic leak (AL) following a colorectal resection. While various recommendations have been proposed to prevent this potentially fatal complication, anastomotic leaks still occur. We present a case of an AL resulting in a complicated and fatal outcome. This case demonstrates the importance of high clinical suspicion, early recognition, and immediate management.

JAK Inhibitors for Treatment of Alopecia Areata.

The advancement of genetic and preclinical studies has uncovered the mechanisms involved in the pathogenesis of alopecia areata (AA). The development of targeted therapies using small molecules blocking specific pathways for the treatment of AA is underway. By repurposing Food and Drug Administration-approved small molecule JAK inhibitors as treatments for AA, it has been demonstrated that JAK inhibitors can effectively reverse hair loss in patients with moderate to severe AA. In this review, we summarize and discuss the current preclinical and clinical studies on JAK inhibitors, as well as the prospects of using JAK inhibitors for the treatment of AA.

Automated Separation of Uranium and Plutonium from Environmental Swipe Samples for Multiple Collector Inductively Coupled Plasma Mass Spectrometry.

A fully automated method for the separation of low-concentration uranium from plutonium in environmental swipe samples has been developed. The offline chromatography system features renewable 1 mL Eichrom TEVA and UTEVA column generation from bulk resin slurry. Discrete fractions of the separated actinides are delivered into user defined vials for future analysis. Clean room background levels were achieved outside of a cleanroom environment with this method. Purification of uranium and plutonium from various sample matrixes and at various concentrations was successful. Major and minor isotope ratios for both elements were measured via multiple collector inductively coupled plasma mass spectrometry and were in good agreement with certified reference values. Validation of the separation method was conducted on archived environmental samples and agreed with values previously reported using standard column chemistry.

Antileukemic Efficacy of BET Inhibitor in a Preclinical Mouse Model of MLL-AF4+ Infant ALL.

MLL-rearranged acute lymphoblastic leukemia (ALL) occurring in infants is a rare but very aggressive leukemia, typically associated with a dismal prognosis. Despite the development of specific therapeutic protocols, infant patients with MLL-rearranged ALL still suffer from a low cure rate. At present, novel therapeutic approaches are urgently needed. Recently, the use of small molecule inhibitors targeting the epigenetic regulators of the MLL complex emerged as a promising strategy for the development of a targeted therapy. Herein, we have investigated the effects of bromodomain and extra-terminal (BET) function abrogation in a preclinical mouse model of MLL-AF4+ infant ALL using the BET inhibitor I-BET151. We reported that I-BET151 is able to arrest the growth of MLL-AF4+ leukemic cells in vitro, by blocking cell division and rapidly inducing apoptosis. Treatment with I-BET151 in vivo impairs the leukemic engraftment of patient-derived primary samples and lower the disease burden in mice. I-BET151 affects the transcriptional profile of MLL-rearranged ALL through the deregulation of BRD4, HOXA7/HOXA9, and RUNX1 gene networks. Moreover, I-BET151 treatment sensitizes glucocorticoid-resistant MLL-rearranged cells to prednisolone in vitro and is more efficient when used in combination with HDAC inhibitors, both in vitro and in vivo Given the aggressiveness of the disease, the failure of the current therapies and the lack of an ultimate cure, this study paves the way for the use of BET inhibitors to treat MLL-rearranged infant ALL for future clinical applications. Mol Cancer Ther; 17(8); 1705-16. ©2018 AACR.

A unique case of popliteal artery transection after a motorcycle collision.

Popliteal artery injuries may have devastating consequences if not recognized in a timely fashion. The risk of delayed diagnosis of a vascular injury is particularly high in blunt trauma to the lower extremity. We present a case of popliteal artery injury that is unusual in two respects: severity (a complete transection in the setting of a blunt injury) and lack of clinical and radiographic signs of the commonly associated musculoskeletal injuries.