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OBJECTIVE: To compare the impact of a commercial "sleep lotion" versus a placebo control lotion on salivary melatonin and sleep quality. METHODS: The presence of melatonin in the lotion was confirmed and quantitated with High Performance Liquid Chromatography (HPLC). In this randomized, controlled, double-blind crossover trial, undergraduate student participants applied lotions on two separate nights and we quantitated melatonin in saliva samples with enzyme linked immunosorbent assays. We also assessed sleep quality with a modified Pittsburgh Sleep Quality Index. CLINICALTRIALS: gov ID NCT06053385. RESULTS: Sixty-three participants (age 20.5 ± 1.2 years; 81% female) enrolled in the study and provided at least one saliva sample. The sleep lotion contained 0.24 ± 0.01% melatonin (g/100 g lotion) which dramatically impacted salivary melatonin levels, increasing them up to ~1000 fold compared to the placebo lotion (n = 36 participants with all six timepoints). The lotion improved sleep quality in a subsample of the poor sleepers (n = 18). However, the sleep quality in the overall sample was not significantly different on the active versus placebo lotion nights. CONCLUSION: In a small undergraduate student sample, a commercial melatonin-containing lotion improved sleep in those with poor sleep quality. Caution should be taken by consumers using over-the-counter melatonin lotions because the undisclosed dosage can be high and well absorbed by the skin.
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A Continuous Performance Task is an example of a mental stressor which requires vigilance, attention, and effort. We hypothesized that a sympathetic nervous system response would be evident from a resting baseline period to this attention test, and explored if physiological measures were correlated to state and trait anxiety, perceived stress, mindfulness, and performance on the task. In 20 undergraduates, blood pressure and skin conductance increased due to the attention test but heart rate variability did not change. The physiological variables did not correlate to psychological variables; there was a trend of higher perceived stress correlating to lower foil accuracy rate ( p = 0.09). ClinicalTrials.gov ID: NCT06098352.
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ABSTRACT: Fanconi anemia (FA) is an inherited DNA repair disorder characterized by bone marrow (BM) failure, developmental abnormalities, myelodysplasia, leukemia, and solid tumor predisposition. Allogeneic hematopoietic stem cell transplantation (allo-HSCT), a mainstay treatment, is limited by conditioning regimen-related toxicity and graft-versus-host disease (GVHD). Antibody-drug conjugates (ADCs) targeting hematopoietic stem cells (HSCs) can open marrow niches permitting donor stem cell alloengraftment. Here, we report that single dose anti-mouse CD45-targeted ADC (CD45-ADC) facilitated stable, multilineage chimerism in 3 distinct FA mouse models representing 90% of FA complementation groups. CD45-ADC profoundly depleted host stem cell enriched Lineage-Sca1+cKit+ cells within 48 hours. Fanca-/- recipients of minor-mismatched BM and single dose CD45-ADC had peripheral blood (PB) mean donor chimerism >90%; donor HSCs alloengraftment was verified in secondary recipients. In Fancc-/- and Fancg-/- recipients of fully allogeneic grafts, PB mean donor chimerism was 60% to 80% and 70% to 80%, respectively. The mean percent donor chimerism in BM and spleen mirrored PB results. CD45-ADC-conditioned mice did not have clinical toxicity. A transient <2.5-fold increase in hepatocellular enzymes and mild-to-moderate histopathological changes were seen. Under GVHD allo-HSCT conditions, wild-type and Fanca-/- recipients of CD45-ADC had markedly reduced GVHD lethality compared with lethal irradiation. Moreover, single dose anti-human CD45-ADC given to rhesus macaque nonhuman primates on days -6 or -10 was at least as myeloablative as lethal irradiation. These data suggest that CD45-ADC can potently promote donor alloengraftment and hematopoiesis without significant toxicity or severe GVHD, as seen with lethal irradiation, providing strong support for clinical trial considerations in highly vulnerable patients with FA.
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Anemia de Fanconi , Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Inmunoconjugados , Antígenos Comunes de Leucocito , Animales , Anemia de Fanconi/terapia , Ratones , Enfermedad Injerto contra Huésped/patología , Inmunoconjugados/farmacología , Inmunoconjugados/uso terapéutico , Acondicionamiento Pretrasplante/métodos , Trasplante Homólogo , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
A variety of mouse models for Down syndrome (Trisomy 21) have been created to test hypotheses about the correlation of phenotypes to gene content and copy number. Ts1Rhr mice are trisomic for a region on mouse chromosome 16 that is homologous to 5.3 Mb of human chromosome 21. Ms1Rhr mice are monosomic for this region. Magnetic Resonance Imaging (MRI) has revealed characteristic volumetric changes in the brains of humans with Down syndrome such as reductions in the cerebellum, hippocampus, and brain stem, and increases in the ventricles and thalamus. We used MRI with region of interest analysis to measure the volume of the thalamus and hypothalamus in Ts1Rhr, Ms1Rhr, and euploid control mice (n = 10-11 per group). Ts1Rhr mice had a 6.6% reduction and Ms1Rhr mice had an 8.2% reduction in the volume of the thalamus. Ts1Rhr and Ms1Rhr hypothalamic volumes were equivalent to controls. Conflicting data in mouse models show a lack of clarity on causative roles of regions homologous to human chromosome 21 in phenotypes related to the thalamus and hypothalamus in Down syndrome.
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Hematopoietic stem cell (HSC) gene therapy has curative potential; however, its use is limited by the morbidity and mortality associated with current chemotherapy-based conditioning. Targeted conditioning using antibody-drug conjugates (ADC) holds promise for reduced toxicity in HSC gene therapy. Here we test the ability of an antibody-drug conjugate targeting CD117 (CD117-ADC) to enable engraftment in a non-human primate lentiviral gene therapy model of hemoglobinopathies. Following single-dose CD117-ADC, a >99% depletion of bone marrow CD34 + CD90 + CD45RA- cells without lymphocyte reduction is observed, which results are not inferior to multi-day myeloablative busulfan conditioning. CD117-ADC, similarly to busulfan, allows efficient engraftment, gene marking, and vector-derived fetal hemoglobin induction. Importantly, ADC treatment is associated with minimal toxicity, and CD117-ADC-conditioned animals maintain fertility. In contrast, busulfan treatment commonly causes severe toxicities and infertility in humans. Thus, the myeloablative capacity of single-dose CD117-ADC is sufficient for efficient engraftment of gene-modified HSCs while preserving fertility and reducing adverse effects related to toxicity in non-human primates. This targeted conditioning approach thus provides the proof-of-principle to improve risk-benefit ratio in a variety of HSC-based gene therapy products in humans.
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Trasplante de Células Madre Hematopoyéticas , Inmunoconjugados , Animales , Busulfano/farmacología , Terapia Genética/métodos , Trasplante de Células Madre Hematopoyéticas/métodos , Células Madre Hematopoyéticas , Inmunoconjugados/farmacología , Proteínas Proto-Oncogénicas c-kit/inmunología , Proteínas Proto-Oncogénicas c-kit/uso terapéutico , Macaca mulatta/inmunologíaRESUMEN
BACKGROUND: Physician burnout increased during the COVID-19 pandemic. OBJECTIVE: To evaluate the effectiveness of a multimodal workplace intervention designed to reduce hospitalist burnout. DESIGN: Participants and setting: Our intervention group was composed of internal medicine hospitalists at Providence Portland Medical Center (64 providers including 58 physicians and 6 nurse practitioners). Our control was composed of internal medicine hospitalists at Providence St Vincent's Hospital (59 physicians and 6 nurse practitioners). MEASUREMENTS: Two surveys were given during, before, and after a 12-month intervention period (October 2020 and again in October 2021). Surveys included demographics, job satisfaction, the Maslach Burnout Inventory, the Pandemic Experiences Survey, and 2 questions about leaving the job. INTERVENTIONS: Three hospitalists designated as wellness warriors created weekly COVID group meetings, providing up-to-date information about COVID-19 infection rates, treatments, and work-flow changes. Discussions included coping and vaccine hesitancy, difficult case debriefs, and intensive care unit updates. Individual coaching was also offered. Meeting minutes were taken and sessions were recorded for asynchronous access. RESULTS: No site differences in burnout or job satisfaction were evident pre-intervention. Post-intervention, the intervention group reported 32% burnout while controls reported 56% (p = .024). Forty-eight percent of the intervention group reported high wellness support vs. 0% of the controls (< .001). Intervention participants attributed 44% of wellness support to Providence alone, vs. controls at 12% (< .001). Regressions controlling sex, work hours, experience, race, and children in the home showed the intervention's positive effects on burnout and job satisfaction remained significant (all p < .02). LIMITATIONS: For privacy reasons, all survey responses were anonymous, meaning that individual pre-post changes could not be tracked. CONCLUSION: We believe the intervention resulted in substantial burnout prevention and is feasible for adoption in most hospitals and clinics.
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Agotamiento Profesional , COVID-19 , Médicos Hospitalarios , Niño , Humanos , COVID-19/epidemiología , COVID-19/prevención & control , Pandemias , Agotamiento Profesional/epidemiología , Agotamiento Profesional/prevención & control , Encuestas y Cuestionarios , Satisfacción en el TrabajoRESUMEN
Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a potentially curative treatment of patients with nonmalignant or malignant blood disorders. Its success has been limited by graft-versus-host disease (GVHD). Current systemic nontargeted conditioning regimens mediate tissue injury and potentially incite and amplify GVHD, limiting the use of this potentially curative treatment beyond malignant disorders. Minimizing systemic nontargeted conditioning while achieving alloengraftment without global immune suppression is highly desirable. Antibody-drug-conjugates (ADCs) targeting hematopoietic cells can specifically deplete host stem and immune cells and enable alloengraftment. We report an anti-mouse CD45-targeted-ADC (CD45-ADC) that facilitates stable murine multilineage donor cell engraftment. Conditioning with CD45-ADC (3 mg/kg) was effective as a single agent in both congenic and minor-mismatch transplant models resulting in full donor chimerism comparable to lethal total body irradiation (TBI). In an MHC-disparate allo-HSCT model, pretransplant CD45-ADC (3 mg/kg) combined with low-dose TBI (150 cGy) and a short course of costimulatory blockade with anti-CD40 ligand antibody enabled 89% of recipients to achieve stable alloengraftment (mean value: 72%). When CD45-ADC was combined with pretransplant TBI (50 cGy) and posttransplant rapamycin, cyclophosphamide (Cytoxan), or a JAK inhibitor, 90% to 100% of recipients achieved stable chimerism (mean: 77%, 59%, 78%, respectively). At a higher dose (5 mg/kg), CD45-ADC as a single agent was sufficient for rapid, high-level multilineage chimerism sustained through the 22 weeks observation period. Therefore, CD45-ADC has the potential utility to confer the benefit of fully myeloablative conditioning but with substantially reduced toxicity when given as a single agent or at lower doses in conjunction with reduced-intensity conditioning.
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Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Inmunoconjugados , Animales , Quimerismo , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Inmunoconjugados/farmacología , Inmunoconjugados/uso terapéutico , Inmunoconjugados/toxicidad , Ratones , Acondicionamiento Pretrasplante/métodosRESUMEN
BACKGROUND: The interest of medical students and surgery residents in global surgery continues to grow. Few studies have examined how the presence of global surgery opportunities influences an applicant's decision to choose a surgical training program. We designed a survey to examine the interest in global surgery among general surgery residency applicants and the influence of a global surgery rotation on a general surgery residency applicant's rank list. METHODS: In March 2019, an online 20-question qualitative survey was administered to all general surgery applicants to a single academic institution. Results were stratified into two applicant groups; applicants from domestic or international medical schools. The survey was designed to capture demographic information, previous global rotations or experiences, future interest in global surgery opportunities, and the importance of global surgery in choosing a residency program. RESULT: s: A total of 179 (21% response rate) applicants completed the entire survey. Of the respondents 81% were interested in a global surgery rotation during residency, 56% considered a global surgery opportunity as moderately to extremely important to their residency rankings, 71% said they would rank a residency higher if it had a funded global surgery program compared to one without funding and 58% of the surveyed applicants were interested in incorporating global surgery into their future career. CONCLUSIONS: Global surgery opportunities are important to some general surgery residency applicants. A majority of applicants believe a funded global surgery would positively influence their rank list. As residency programs train residents for their future careers greater consideration needs to be given to developing global surgery opportunities.
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Selección de Profesión , Educación de Postgrado en Medicina , Cirugía General , Salud Global , Estudiantes de Medicina/psicología , Adulto , Femenino , Humanos , Masculino , Estudiantes de Medicina/estadística & datos numéricos , Adulto JovenRESUMEN
Cytokines that signal through the JAK-STAT pathway, such as interferon-γ (IFN-γ) and common γ chain cytokines, contribute to the destruction of insulin-secreting ß cells by CD8+ T cells in type 1 diabetes (T1D). We previously showed that JAK1/JAK2 inhibitors reversed autoimmune insulitis in non-obese diabetic (NOD) mice and also blocked IFN-γ mediated MHC class I upregulation on ß cells. Blocking interferons on their own does not prevent diabetes in knockout NOD mice, so we tested whether JAK inhibitor action on signaling downstream of common γ chain cytokines, including IL-2, IL-7 IL-15, and IL-21, may also affect the progression of diabetes in NOD mice. Common γ chain cytokines activate JAK1 and JAK3 to regulate T cell proliferation. We used a JAK1-selective inhibitor, ABT 317, to better understand the specific role of JAK1 signaling in autoimmune diabetes. ABT 317 reduced IL-21, IL-2, IL-15 and IL-7 signaling in T cells and IFN-γ signaling in ß cells, but ABT 317 did not affect GM-CSF signaling in granulocytes. When given in vivo to NOD mice, ABT 317 reduced CD8+ T cell proliferation as well as the number of KLRG+ effector and CD44hiCD62Llo effector memory CD8+ T cells in spleen. ABT 317 also prevented MHC class I upregulation on ß cells. Newly diagnosed diabetes was reversed in 94% NOD mice treated twice daily with ABT 317 while still on treatment at 40 days and 44% remained normoglycemic after a further 60 days from discontinuing the drug. Our results indicate that ABT 317 blocks common γ chain cytokines in lymphocytes and interferons in lymphocytes and ß cells and are thus more effective against diabetes pathogenesis than IFN-γ receptor deficiency alone. Our studies suggest use of this class of drug for the treatment of type 1 diabetes.
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Diabetes Mellitus Tipo 1/inmunología , Interferón gamma/inmunología , Subunidad gamma Común de Receptores de Interleucina/inmunología , Janus Quinasa 1/antagonistas & inhibidores , Inhibidores de las Cinasas Janus/farmacología , Animales , Células Secretoras de Insulina/efectos de los fármacos , Células Secretoras de Insulina/inmunología , Inhibidores de las Cinasas Janus/farmacocinética , Masculino , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Transducción de Señal/efectos de los fármacos , Bazo/inmunologíaRESUMEN
INTRODUCTION: Many medical schools offer M4 boot camps to improve students' preparedness for surgical residencies. For three consecutive years, we investigated the impact of medical school boot camps on intern knot-tying and suturing skills when measured at the start of residency. METHODS: Forty-two interns completed questionnaires regarding their boot camp experiences. Their performance on knot-tying and suturing exercises was scored by three surgeons blinded to the questionnaire results. A comparison of these scores of interns with or without boot camp experiences was performed and statistical analysis applied. RESULTS: 26 of 42 (62%) interns reported boot camp training. There were no differences in scores between interns with or without a M4 boot camp experience for suturing [9.6(4.6) vs 9.8(4.1), p < 0.908], knot-tying [9.1(3.6) vs 8.4(4.1), p = 0.574], overall performance [2.0(0.6) vs 1.9(0.7), p = 0.424], and quality [2.0(0.6) vs 1.9(0.7), p = 0.665]) (mean(SD)). CONCLUSIONS: We could not demonstrate a statistically significant benefit in knot-tying and suturing skills of students who enrolled in M4 boot camp courses as measured at the start of surgical residency.
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Competencia Clínica , Educación de Postgrado en Medicina/métodos , Evaluación Educacional/métodos , Técnicas de Sutura/educación , Femenino , Humanos , Internado y Residencia , Masculino , Reproducibilidad de los Resultados , Facultades de Medicina , Encuestas y Cuestionarios , Adulto JovenRESUMEN
BACKGROUND: Graduating general surgery residents are required to pass the FES exam for ABS certification. Trainees and surgery educators are interested in defining the most effective methods of exam preparation. Our aim is to define trainee perceptions, performance, and the most effective preparation methods regarding the FES exam. METHODS: General surgery residents from a single institution who completed the FES exam were identified. All participated in a flexible endoscopy rotation, and all had access to an endoscopy simulator. Residents were surveyed regarding preparation methods and exam difficulty. Descriptive statistics and a Kruskal-Wallis test were used. RESULTS: A total of 26 trainees took the FES exam with a first-time pass rate of 96.2%. Of 26 surveys administered, 21 were completed. Twenty trainees (76.9%) participated in a dedicated endoscopy curriculum. Scores were not different among those who received dedicated curricular instruction compared to those who did not (547 [IQR 539-562.5] vs. 516 [484.5-547], p = 0.1484; 535.5 [468.5-571] vs. 519 [464.75-575], p = 0.9514). Written exam difficulty was rated as 5.5 on a 10-point Likert scale, and 85.7% felt it was a fair assessment of endoscopy knowledge; skills exam difficulty was rated as 7, and 71% felt it was a fair assessment of endoscopy skills. Online FES modules, the endoscopy clinical rotation, and an exam preparation session with a faculty member were most effective for written exam preparation. The most effective skills exam preparation methods were independent simulator practice, the endoscopy clinical rotation, and a preparation session with a faculty member. The most difficult skills were loop reduction and retroflexion. Skill decay did not appear to be significant. CONCLUSIONS: A clinical endoscopy rotation, a method for independent skills practice, and faculty-mediated exam instruction appear to be effective exam preparation methods. When these are present, trainees report minimal need for dedicated exam preparation time prior to taking the FES exam.
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Competencia Clínica/normas , Endoscopía/educación , Humanos , Internado y Residencia , Encuestas y CuestionariosRESUMEN
OBJECTIVE: Mindfulness-based interventions have been shown to have psychological benefits in college students. We explored the effects of an academic Seminar on Compassion on student psychological health. PARTICIPANTS: Forty-one participants (14 male, 27 female, mean age 19.8 ± 1.4 years) were assessed pre- and post- spring semesters 2013 and 2014. METHODS: Students were randomized to the Seminar on Compassion or a wait-list control group. Participants completed self-report measures on anxiety, depression, perceived stress, self-compassion, compassion and mindfulness. Salivary alpha-amylase was also assessed. RESULTS: At baseline, self-compassion and mindfulness were negatively correlated with depression, anxiety, and perceived stress. There were significant changes between the intervention and control group from Time 1 to Time 2 in mindfulness, self-compassion, compassion, and salivary alpha-amylase; however, there were no significant changes in depression, anxiety, and perceived stress. CONCLUSIONS: The course was effective in increasing mindfulness, self-compassion and compassion, and decreasing a salivary marker of stress.
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Empatía , Salud Mental/estadística & datos numéricos , Atención Plena/educación , Estudiantes/psicología , Adolescente , Ansiedad/epidemiología , Depresión/epidemiología , Femenino , Humanos , Masculino , Autoinforme , Estrés Psicológico/epidemiología , Universidades , Adulto Joven , alfa-Amilasas/análisisRESUMEN
BACKGROUND: Anti-cytokine therapies such as adalimumab, tocilizumab, and the small molecule JAK inhibitor tofacitinib have proven that cytokines and their subsequent downstream signaling processes are important in the pathogenesis of rheumatoid arthritis. Tofacitinib, a pan-JAK inhibitor, is the first approved JAK inhibitor for the treatment of RA and has been shown to be effective in managing disease. However, in phase 2 dose-ranging studies tofacitinib was associated with dose-limiting tolerability and safety issues such as anemia. Upadacitinib (ABT-494) is a selective JAK1 inhibitor that was engineered to address the hypothesis that greater JAK1 selectivity over other JAK family members will translate into a more favorable benefit:risk profile. Upadacitinib selectively targets JAK1 dependent disease drivers such as IL-6 and IFNγ, while reducing effects on reticulocytes and natural killer (NK) cells, which potentially contributed to the tolerability issues of tofacitinib. METHODS: Structure-based hypotheses were used to design the JAK1 selective inhibitor upadacitinib. JAK family selectivity was defined with in vitro assays including biochemical assessments, engineered cell lines, and cytokine stimulation. In vivo selectivity was defined by the efficacy of upadacitinib and tofacitinib in a rat adjuvant induced arthritis model, activity on reticulocyte deployment, and effect on circulating NK cells. The translation of the preclinical JAK1 selectivity was assessed in healthy volunteers using ex vivo stimulation with JAK-dependent cytokines. RESULTS: Here, we show the structural basis for the JAK1 selectivity of upadacitinib, along with the in vitro JAK family selectivity profile and subsequent in vivo physiological consequences. Upadacitinib is ~ 60 fold selective for JAK1 over JAK2, and > 100 fold selective over JAK3 in cellular assays. While both upadacitinib and tofacitinib demonstrated efficacy in a rat model of arthritis, the increased selectivity of upadacitinib for JAK1 resulted in a reduced effect on reticulocyte deployment and NK cell depletion relative to efficacy. Ex vivo pharmacodynamic data obtained from Phase I healthy volunteers confirmed the JAK1 selectivity of upadactinib in a clinical setting. CONCLUSIONS: The data presented here highlight the JAK1 selectivity of upadacinitinib and supports its use as an effective therapy for the treatment of RA with the potential for an improved benefit:risk profile.
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An interdisciplinary, intensive, and experiential course on Neuroscience of Meditation was designed to fulfill a general education science requirement. Class activities included lecture, class discussion of the textbook and scholarly articles, laboratory experimentation, and practicing 15 different forms of meditation. Laboratory investigations included sheep brain dissection, physiological measurements of the autonomic nervous system, electroencephalogram, salivary enzyme assays, attention testing, and psychological questionnaires. The Determinants of Meditation Practice Inventory (DMPI) and My Attitude Toward Science (MATS) scales were administered at the beginning and conclusion of the course; barriers to meditation were reduced and positive attitudes toward school science were increased. Comments on course evaluations praised this incorporation of contemplative pedagogy into a neuroscience course.
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Smith, CA, Olson, BK, Olson, LA, Chimera, NJ, and Warren, M. Comparison of female collegiate athletes and college age cohort in tuck jump assessment. J Strength Cond Res 31(4): 1048-1054, 2017-The tuck jump assessment (TJA) is a plyometric jumping assessment with 10 flaw criteria against which technique is assessed over a 10-second interval. The TJA has been reported as a tool for identifying neuromuscular deficits that increase risk for anterior cruciate ligament injury, but group specific data on female TJA scores are limited. No cut point has been developed for groups with different activity levels or participation in athletics. This study investigated the association between TJA score and athletic participation in college-aged females. One hundred twenty-one females (53 collegiate athletes and 68 college students) completed the TJA. TJA score was the sum of flaws for the 10 criteria observed, and the number of jumps was recorded. Poisson regression was used to assess the association between TJA score and number of jumps. The association between each of the 10 flaws between groups was assessed with the chi-square test. No significant association was found between groups for TJA score (mean ± SD: 4.66 ± 1.07 athletes; 5.45 ± 1.05 college cohort; p = 0.06; ß = 0.82). Athletes jumped significantly more times (12.23 ± 1.04 athletes; 9.35 ± 1.04 college cohort). Athletes had a lower proportion of 2 flaws: "thighs do not reach parallel" and "pause between jumps." Lower statistical power may limit interpretation of the remaining flaws. The lack of control of the number of jumps may impact TJA score. To improve the TJA usefulness on the field and clinic, the protocol may need to standardize the number of jumps.
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Lesiones del Ligamento Cruzado Anterior/fisiopatología , Atletas , Prueba de Esfuerzo/métodos , Adolescente , Fenómenos Biomecánicos , Femenino , Humanos , Universidades , Adulto JovenRESUMEN
Undergraduate courses in biopsychology, neuroscience, and physiology often include laboratory exercises that examine responses to stimulation of the sympathetic nervous system with measurements of heart rate, blood pressure, or galvanic skin levels (sweat response). A newer bioindicator of the sympathetic nervous system is salivary alpha-amylase (sAA) measured with a colorimetic enzyme assay. Undergraduate students successfully measured a rise in sAA due to the stress of giving a class presentation (n=13). Students were enthusiastic to measure a physiological response to a real-life anxiety-producing situation. We describe potential difficulties in the assay and our adaptations to the manufacturer's protocol to make it more feasible in the undergraduate setting.
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OBJECTIVE: In lupus nephritis, tubulointerstitial inflammation (TII) is associated with in situ adaptive immune cell networks that amplify local tissue damage. Since conventional therapy appears ineffective for severe TII, and these patients often progress to renal failure, understanding in situ mechanisms might reveal new therapeutic targets. This study was undertaken to assess whether dysregulated apoptotic regulators maintain local adaptive immunity and drive inflammation in TII. METHODS: This study utilized novel computational approaches that, when applied to multicolor confocal images, quantified apoptotic regulator protein expression in selected lymphocyte subsets. This approach was validated using laser-capture microdissection (LCM) coupled to quantitative polymerase chain reaction (qPCR). Furthermore, the consequences of dysregulated apoptotic mediator expression were explored in a murine model of lupus nephritis. RESULTS: Analyses of renal biopsy tissue from patients with lupus nephritis and those with mixed cellular renal allograft rejection revealed that the B cell lymphoma 2 protein (Bcl-2) was frequently expressed in infiltrating lymphocytes, whereas expression of myeloid cell leukemia 1 was low. In contrast, the reciprocal pattern of expression was observed in tonsil germinal centers. These results were consistent with RNA expression data obtained using LCM and qPCR. Bcl-2 was also highly expressed in tubulointerstitial infiltrates in (NZB × NZW)F1 (NZB/NZW) mice. Furthermore, treatment of NZB/NZW mice with ABT-199, a selective oral inhibitor of Bcl-2, prolonged survival and prevented proteinuria and development of TII in a lupus prevention model. Interestingly, glomerular immune complexes were partially ameliorated by ABT-199 treatment, and serum anti-double-stranded DNA antibody titers were unaffected. CONCLUSION: These data demonstrate that Bcl-2 is an attractive therapeutic target in patients with lupus nephritis who manifest TII.
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Apoptosis , Nefritis Lúpica/metabolismo , Linfocitos/metabolismo , Nefritis Intersticial/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Inmunidad Adaptativa/inmunología , Animales , Complejo Antígeno-Anticuerpo/efectos de los fármacos , Complejo Antígeno-Anticuerpo/inmunología , Complejo Antígeno-Anticuerpo/metabolismo , Proteína 11 Similar a Bcl2/genética , Proteína 11 Similar a Bcl2/metabolismo , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Femenino , Centro Germinal/metabolismo , Rechazo de Injerto/inmunología , Rechazo de Injerto/metabolismo , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Inflamación , Riñón/efectos de los fármacos , Riñón/inmunología , Riñón/metabolismo , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/inmunología , Glomérulos Renales/metabolismo , Trasplante de Riñón , Captura por Microdisección con Láser , Nefritis Lúpica/inmunología , Linfocitos/inmunología , Ratones , Ratones Endogámicos NZB , Microscopía Confocal , Microscopía Fluorescente , Proteína 1 de la Secuencia de Leucemia de Células Mieloides/genética , Proteína 1 de la Secuencia de Leucemia de Células Mieloides/metabolismo , Nefritis Intersticial/inmunología , Tonsila Palatina , Reacción en Cadena de la Polimerasa , Proteínas Proto-Oncogénicas c-bcl-2/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/genética , Sulfonamidas/farmacologíaRESUMEN
The anthracycline chemotherapeutic agent doxorubicin is converted by the enzyme carbonyl reductase 1 (CBR1) into its cardiotoxic metabolite doxorubicinol. Cbr1+/- mice have been shown to be protected from doxorubicin-induced cardiotoxicity, and the inhibition of CBR1 activity may be a useful means of ameliorating the side effects of doxorubicin in patients undergoing chemotherapy. Because reduced conversion to doxorubicinol increases circulating levels of the more effective parent drug doxorubicin, it was hypothesized that therapeutic efficacy against tumors might also be enhanced. Cbr1+/- mice were bred to mice transgenic for the polyomavirus middle T antigen (PyVT) to create offspring with palpable mammary tumors. Latency to initial tumor formation was similar in Cbr1+/- and Cbr1+/+ animals. Tumor regression was improved in Cbr1+/- animals, but only in male mice. Western blotting showed a marked sex difference in protein levels, with a much higher expression of Cbr1 in the female kidney and liver. Thus, the combined effects of a naturally low expression and the heterozygous Cbr1 null allele seem to have enhanced tumor regression in Cbr1+/- males. Future efforts to design a clinical CBR1 inhibitor to protect patients from the cardiac side effects of doxorubicin treatment should evaluate the effect of sex on anticancer efficacy.
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Oxidorreductasas de Alcohol/genética , Antibióticos Antineoplásicos/efectos adversos , Doxorrubicina/efectos adversos , Oxidorreductasas de Alcohol/metabolismo , Animales , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/farmacología , Doxorrubicina/análogos & derivados , Doxorrubicina/metabolismo , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Femenino , Riñón/efectos de los fármacos , Riñón/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Neoplasias Mamarias Animales/tratamiento farmacológico , Ratones , Ratones Mutantes , Ratones TransgénicosRESUMEN
Immunological responses to protect against excessive inflammation can be regulated by the central nervous system through the cholinergic anti-inflammatory pathway wherein acetylcholine released from vagus nerves can inhibit inflammatory cytokines. Although a role for the α7 nicotinic acetylcholine receptor (α7 nAChR) in mediating this pathway has been suggested, pharmacological modulation of the pathway by selective agonists remains to be further elucidated. In this study, the role of α7 nAChRs in the regulation of TNF-α release was investigated using high affinity and selective α7 nAChR agonists in mouse peritoneal macrophage and human whole blood in vitro, and in mouse serum in vivo. In mouse peritoneal macrophages, LPS-induced TNF-α release in vitro was inhibited by a selective α7 nAChR agonist, A-833834 (5-[6-(5-Methyl-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-pyridazin-3-yl]-1H-indole), and that effect was attenuated by α7 nAChR antagonist methyllycaconitine. The inhibitory effect of A-833834 on LPS-induced TNF-α release was also observed in human whole blood in vitro. I.v. LPS-induced TNF-α release in mouse serum was attenuated following i.p. administration of A-833834. Similarly, i.v. LPS-induced TNF-α release in mouse serum was also attenuated following i.p. administration of A-585539, another α7 nAChR agonist with limited brain penetration, suggesting that these effects are mediated by peripheral α7 nAChRs. A-833834 was also efficacious in suppressing TNF-α release in mouse serum following oral administration in zymosan-induced peritonitis. These studies collectively demonstrate that selectively targeting α7 nAChRs could offer a novel therapeutic modality to treat acute and chronic inflammatory disease states.
Asunto(s)
Agonistas Nicotínicos/sangre , Agonistas Nicotínicos/farmacología , Receptores Nicotínicos/fisiología , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Citocinas/antagonistas & inhibidores , Citocinas/sangre , Citocinas/metabolismo , Femenino , Humanos , Mediadores de Inflamación/agonistas , Mediadores de Inflamación/antagonistas & inhibidores , Mediadores de Inflamación/sangre , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/metabolismo , Macrófagos Peritoneales/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Oocitos , Peritonitis/tratamiento farmacológico , Peritonitis/inmunología , Peritonitis/patología , Unión Proteica/efectos de los fármacos , Unión Proteica/inmunología , Receptores Nicotínicos/sangre , Receptores Nicotínicos/metabolismo , Factor de Necrosis Tumoral alfa/sangre , Xenopus laevis , Receptor Nicotínico de Acetilcolina alfa 7RESUMEN
Down syndrome (trisomy 21) is associated with reduced bone density in humans, but it is unclear whether this is due to specific effects of chromosome 21 genes or lifestyle factors. Mouse models with aneuploidy of segments of mouse chromosome 16 that are homologous to human chromosome 21 can be used to elucidate the mechanism by which Down syndrome phenotypes arise. Ts1Rhr and Ms1Rhr mice are trisomic and monosomic, respectively, for the hypothesized "Down syndrome critical region" containing approximately 33 genes. We assessed the skeletons of these mice from 3 to 16 weeks of age using dual X-ray absorptiometry. Ts1Rhr mice were unexpectedly similar to normal controls, showing that a larger region of trisomy is necessary to recapitulate the Down syndrome phenotype. Ms1Rhr mice, in contrast, showed decreases in weight, bone mineral content, bone mineral density, and bone area from weaning to adulthood. Regional bone density was also decreased in the femur, tibia, and lower lumbar spine. The microarchitecture of 3 week old Ms1Rhr femurs was then analyzed using µCT. Volumetric density, total tissue volume, bone volume, and bone fraction were all reduced in both cortical and trabecular bone. Ms1Rhr trabeculae were thinner and had decreased connectivity. A 31.5% reduction in the level of insulin-like growth factor I in the serum was found, and we hypothesize that this is responsible for the bone density phenotype. We discuss bone-related genes in the region and propose that humans with distal chromosome 21 deletions may exhibit reduced bone density.