Marked Response Research Articles

Evaluation of the therapeutic effect of PUVA therapy in chronic vitiligo patients

Introduction: Vitiligo is an acquired pigmentary disease, characterized by depigmented macules or patches which distracts the psychosocial behaviour of patients. Various therapeutic modalities have been admintered over time with variable outcome. Psoralen followed by irradiation with UV-A (PUVA) is one of the effective treatment methods for chronic vitiligo cases who had failed to react the other treatment methods. Aim: To assess the efficacy of therapeutic effect of oral psoralen followed by ultraviolet-A irradiation (PUVA) therapy for chronic vitiligo cases. Materials and Methods: A total 36 patients of both sexes with acral, acrofacial and generalised vitiligo with more than 20% spread over body, between age group 15-60 years were recruited. 8 MOP tablets are given with empty stomach according to body weight. All cases were undergone to UVA exposure artificial phototherapy chamber starting with a dose of 4 J/m2 (Dosage depends on Fitzpatrick skin type) over whole body after 90 min. Clinical response was evaluated as marked response rate. Results: Generalised vitiligo was seen in 66.6% cases, acral was seen in 16.6% cases, acrofacial was seen in 16.6% cases. Erythema was the common side effect followed by xerosis, pruritus, nausea and vomiting, bulla and burning. The mean duration of treatment was 14.52 months with mean no of sessions is 62.18. Conclusion: PUVA therapy is a well-established treatment modality for vitiligo with very minimal risk of malignancies and has less side effects like erythema and xerosis. PUVA is safe, effective, and cosmetically acceptable. Generalized vitiligo was responded well to the treatment. Keywords: Psoralen and ultraviolet A (PUVA), Generalised vitiligo, Acral vitiligo, Acrofacial vitiligo.

High-Dose Thymidine Infusions in Patients With Leukemia and Lymphoma

We have recently explored the cytokinetic effects of thymidine given by continuous infusion in an animal model and have demonstrated an arrest of cell cycle traverse in rapidly proliferating tissues. Similar biologic effects have been observed in clinical trials upon infusing thymidine at a dose rate of 75 g/sq m/day. Fifteen courses of at least 5 days in duration have been administered to 11 patients with either leukemia or lymphoma. Steady-state serum thymidine levels were achieved in the range of 1-2 mM and the serum half-life of thymidine was approximately 90 min upon completion of the infusion. The associated toxicity included myelosuppression, headache, anorexia, nausea, vomiting, and diarrhea. The antitumor effect was dependent on the disease being treated and the percentage of blasts in the peripheral blood. Three patients with AML and three patients with T-cell leukemia responded to the thymidine infusions with an abrupt decrease in peripheral blast count. In contrast, no response was observed in two patients with poorly differentiated lymphocytic lymphoma of B-cell origin in a leukemic phase. One patient with T-cell leukemia had a marked clinical response with resolution of hepatosplenomegaly and another patient with the same diagnosis had partial clearing of the bone marrow. The cytokinetic effect on the bone marrow or peripheral blasts was monitored by microfluorimetry and labeling with precursors to monitor DNA synthesis. These data indicate an arrest of DNA synthesis during the period of thymidine infusion. The clinical effects observed with the high-dose thymidine infusions are correlated with measurements of thymidine kinase and phosphorylase levels detected in the blasts obtained from the leukemic patients. The relative ratios of thymidine kinase to phosphorylase are predictive of cellular sensitivity to the cytokinetic effects of thymidine.

Fms-Like Tyrosine Kinase 3-Independent Dendritic Cells Are Major Mediators of Th2 Immune Responses in Allergen-Induced Asthmatic Mice.

Dendritic cells (DCs) are the main mediators of Th2 immune responses in allergic asthma, and Fms-like tyrosine kinase 3 ligand (Flt3L) is an important growth factor for the development and homeostasis of DCs. This study identified the DC populations that primarily cause the initiation and development of allergic lung inflammation using Fms-like tyrosine kinase 3 (Flt3) knockout (KO) mice with allergen-induced allergic asthma. We observed type 2 allergic lung inflammation with goblet cell hyperplasia in Flt3 KO mice, despite a significant reduction in total DCs, particularly CD103+ DCs, which was barely detected. In addition, bone marrow-derived dendritic cells (BMDCs) from Flt3 KO mice directed Th2 immune responses in vitro, and the adoptive transfer of these BMDCs exacerbated allergic asthma with more marked Th2 responses than that of BMDCs from wild-type (WT) mice. Furthermore, we found that Flt3L regulated the in vitro expression of OX40 ligand (OX40L) in DCs, which is correlated with DC phenotype in in vivo models. In conclusion, we revealed that Flt3-independent CD11b+ DCs direct Th2 responses with the elevated OX40L and are the primary cause of allergic asthma. Our findings suggest that Flt3 is required to control type 2 allergic inflammation.

Aging- and gender-related modulation of RAAS: potential implications in COVID-19 disease.

Coronavirus disease 2019 (COVID-19) is a new infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19 is frequently characterized by a marked inflammatory response with severe pneumonia and respiratory failure associated with multiorgan involvement. Some risk factors predispose patients to develop a more severe infection and to an increased mortality; among them, advanced age and male gender have been identified as major and independent risk factors for COVID-19 poor outcome. The renin-angiotensin-aldosterone system (RAAS) is strictly involved in COVID-19 because angiotensin converting enzyme 2 (ACE2) is the host receptor for SARS-CoV-2 and also converts pro-inflammatory angiotensin (Ang) II into anti-inflammatory Ang(1–7). In this review, we have addressed the effect of aging and gender on RAAS with emphasis on ACE2, pro-inflammatory Ang II/Ang II receptor 1 axis and anti-inflammatory Ang(1–7)/Mas receptor axis.

Efficacy of intralesional Purified Protein Derivative (PPD) in the treatment of multiple warts

Cutaneous warts are a common dermatological condition caused by the human papillomavirus (HPV) strains 1,2,4 and rarely 57. Although it is a benign condition, it causes disfigurement, pain, discomfort, has a tendency to koebnerize, and can be transmitted to others. This makes adequate and timely treatment important. There are several conventional treatments available which include electrosurgery, cryotherapy and topical medications like imiquimod which show variable responses. Our study evaluated the effectiveness and associated side effects of intralesional PPD for the treatment of multiple warts in a group of 20 patients. This is done as a form of immunotherapy. It was administered at a dosage of 0.1ml into each lesional site every two weeks until the clearance of lesions or up to a maximum of 6 injections. We observed a marked response in 5%, moderate response in 75% and no response in 20 % of our study population with no complications. On comparison to previous similar studies, the accuracy of our observed results for efficacy and observed side effects was reinforced. Hence proves to be a modality of treatment that should be more widely used for the treatment of warts as it is cheap, safe, less stressful and comparatively more effective.

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada).

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.

Histological Study of Human Placenta in Preeclampsia and the Role of Decidual Natural Killer Cells and Macrophages in its Pathogenesis

Background: Preeclampsia (PE) is a hazardous pregnancy condition, if untreated can result in serious complications to the mother and the fetus.Aim of work: Investigating the histological changes of human placenta in PE and the involvement of natural killer (NK) cells and macrophages in its pathogenesis.Subjects and Methods: Forty pregnant women were included; 20 were normal clinically and considered the control (group I) and the other 20 were included in PE (group II). Blood samples were taken just before delivery to assess serum interleukins (IL)-10 and -12 and transforming growth factor (TGF)-β1. Placental sections were stained with H&E, Mallory's trichrome and immunohistochemically with Bcl-2, vascular endothelial growth factor (VEGF), CD56 and CD68. Then morphometric and statistical studies were done.Results: Group II (PE) showed significantly decreased IL-10 and significantly increased IL-12 and TGF-β1 compared to group I. It also presented different histological alterations in placental sections mainly in the chorionic plates and villi. There was significant increase in area percent of collagen fibers and VEGF, and in the number of syncytial knots, CD56 positive decidual NK cells and CD68 positive decidual macrophages. But there was significant decrease in the area percent of Bcl-2 immunoexpression.Conclusion: Preeclampsia is associated with marked inflammatory and immune responses, and decidual NK cells and macrophages are highly imperative cells in its pathogenesis

Impact of neoadjuvant chemotherapy and stereotactic body radiation therapy (SBRT) on R0 resection rate for borderline resectable and locally advanced pancreatic cancer

BackgroundThe role of neoadjuvant stereotactic body radiation therapy (SBRT) in patients with borderline resectable pancreas cancer (BRPC) and locally advanced pancreas cancer (LAPC) remains controversial. MethodsWe retrospectively evaluated BRPC and LAPC patients treated at our institution who underwent 2–3 months of chemotherapy followed by SBRT to a dose of 30–33 Gy. Overall survival (OS) and recurrence-free survival (RFS) were estimated and compared by Kaplan–Meier and log-rank methods. ResultsWe identified 103 (85 BRPC and 18 LAPC) patients treated per our neoadjuvant paradigm between 2011 and 2018, with resectability based on NCCN definitions. Median follow up was 25 months. Of patients completing neoadjuvant therapy, 73 (71%) underwent definitive resection. Seventy-one (97%) patients with definitively resected tumors had R0 resection and 5 (7%) had a complete pathologic response CR to neoadjuvant therapy. The median overall survival (OS) of the cohort was 24 months. Those with a complete or marked pathologic response had significantly better OS than those with a moderate response (41 vs 24 months, p < 0.02) and patients unable to undergo definitive surgery (17 months, p < 0.0003). Six resected patients experienced grade ≥3 surgical complications. ConclusionsNeoadjuvant chemotherapy and SBRT are associated with promising pathologic response rates and R0 resection rates, with acceptable perioperative morbidity.

Is Levodopa Response a Valid Indicator of Parkinson's Disease?

The clinical diagnosis of Parkinson's disease (PD) requires the presence of parkinsonism and supportive criteria that include a clear and dramatic beneficial response to dopaminergic therapy. Our aim was to test the diagnostic criterion of dopaminergic response by evaluating its association with pathologically confirmed diagnoses in a large population of parkinsonian patients. We reviewed clinical data maintained in an electronic medical record from all patients with autopsy data who had been seen in the Movement Disorders Center at Washington University, St. Louis, between 1996 and 2018. All patients with parkinsonism who underwent postmortem neuropathologic examination were included in this analysis. There were 257 unique parkinsonian patients with autopsy-based diagnoses who had received dopaminergic therapy. Marked or moderate response to dopaminergic therapy occurred in 91.2% (166/182) of those with autopsy-confirmed PD, 52.0% (13/25) of those with autopsy-confirmed multiple systems atrophy, 44.4% (8/18) of those with autopsy-confirmed progressive supranuclear palsy, and 1 (1/8) with autopsy-confirmed corticobasal degeneration. Other diagnoses were responsible for the remaining 24 individuals, 9 of whom had a moderate response to dopaminergic therapy. A substantial response to dopaminergic therapy is frequent but not universal in PD. An absent response does not exclude PD. In other neurodegenerative disorders associated with parkinsonism, a prominent response may also be evident, but this occurs less frequently than in PD. © 2020 International Parkinson and Movement Disorder Society.

Insulin inhibits inflammation-induced cone death in retinal detachment

BackgroundRhegmatogenous retinal detachment (RD) involving the macula is a major cause of visual impairment despite high surgical success rate, mainly because of cone death. RD causes the infiltration of activated immune cells, but it is not clear whether and how infiltrating inflammatory cells contribute to cone cell loss.MethodsVitreous samples from patients with RD and from control patients with macular hole were analyzed to characterize the inflammatory response to RD. A mouse model of RD and retinal explants culture were then used to explore the mechanisms leading to cone death.ResultsAnalysis of vitreous samples confirms that RD induces a marked inflammatory response with increased cytokine and chemokine expression in humans, which is closely mimicked by experimental murine RD. In this model, we corroborate that myeloid cells and T-lymphocytes contribute to cone loss, as the inhibition of their accumulation by Thrombospondin 1 (TSP1) increased cone survival. Using monocyte/retinal co-cultures and TSP1 treatment in RD, we demonstrate that immune cell infiltration downregulates rod-derived cone viability factor (RdCVF), which physiologically regulates glucose uptake in cones. Insulin and the insulin sensitizers rosiglitazone and metformin prevent in part the RD-induced cone loss in vivo, despite the persistence of inflammationConclusionOur results describe a new mechanism by which inflammation induces cone death in RD, likely through cone starvation due to the downregulation of RdCVF that could be reversed by insulin. Therapeutic inhibition of inflammation and stimulation of glucose availability in cones by insulin signaling might prevent RD-associated cone death until the RD can be surgically repaired and improve visual outcome after RD.Trial registrationClinicalTrials.govNCT03318588

Development of CpG-adjuvanted stable prefusion SARS-CoV-2 spike antigen as a subunit vaccine against COVID-19

The COVID-19 pandemic is a worldwide health emergency which calls for an unprecedented race for vaccines and treatment. In developing a COVID-19 vaccine, we applied technology previously used for MERS-CoV to produce a prefusion-stabilized SARS-CoV-2 spike protein, S-2P. To enhance immunogenicity and mitigate the potential vaccine-induced immunopathology, CpG 1018, a Th1-biasing synthetic toll-like receptor 9 (TLR9) agonist was selected as an adjuvant candidate. S-2P in combination with CpG 1018 and aluminum hydroxide (alum) was found to be the most potent immunogen and induced high titer of neutralizing antibodies in sera of immunized mice against pseudotyped lentivirus reporter or live wild-type SARS-CoV-2. In addition, the antibodies elicited were able to cross-neutralize pseudovirus containing the spike protein of the D614G variant, indicating the potential for broad spectrum protection. A marked Th1 dominant response was noted from cytokines secreted by splenocytes of mice immunized with CpG 1018 and alum. No vaccine-related serious adverse effects were found in the dose-ranging study in rats administered single- or two-dose regimens of S-2P combined with CpG 1018 alone or CpG 1018 with alum. These data support continued development of CHO-derived S-2P formulated with CpG 1018 and alum as a candidate vaccine to prevent COVID-19 disease.

Abstract 141: Early Cytokine Profile in Selectively Vulnerable Brain Regions, Heart and Serum After Asphyxial Cardiac Arrest vs. Ventricular Fibrillation Cardiac Arrest in Rats

Background: Currently, post-resuscitative care for survivors of cardiac arrest (CA) is independent of the etiology of CA. We have previously reported that asphyxial CA (ACA) results in different injury pattern vs. ventricular fibrillation CA (VFCA). Exploration of the early inflammatory response could identify insult-specific targets for future therapies. We hypothesized that the early systemic and regional neuroinflammatory cytokine response could differ between ACA vs. VFCA. Methods: Adult male rats were subjected to a 10 min ACA, or 10 min VFCA (n=12/group). Naives (n=12) and shams (n=12) subjected to anesthesia and instrumentation but not CA served as controls. Serum, heart and regional brain cytokine signatures were assessed in selectively vulnerable brain regions: cerebellum, cortex, hippocampus and striatum at 3 h. NSE, S100b and troponin T were also assessed. Results: Asphyxiation produced an ~4 min period of severe hypoxemia followed by a no-flow duration of ~ 6±1 min. VF immediately induced no-flow. Brain cytokines in naïve rats were low or undetectable. Shams had a modest effect on selected cytokines. Both ACA and VFCA resulted in robust selected cytokine responses in serum, heart and brain (Fig. 1) with significant regional differences in brain that identified striatum as a selective neuroinflammatory target. No significant differences in cytokines between CA models were observed. NSE, S100b and troponin T were similarly increased after ACA and VFCA vs. naives and/or shams. Conclusions: Both models of CA resulted in marked systemic, heart and brain cytokine response. Biomarkers of organ injury were also comparable. Contrasting prior studies showing greater oxidative stress and brain injury in ACA, but greater hemodynamic impairment after VFCA, systemic and brain cytokine profiles revealed similar increases across insults, suggesting that the cytokine response may not mediate key secondary injury differences between these two CA phenotypes.

Primary colon adenocarcinoma with choriocarcinoma differentiation: a case report and review of the literature

BackgroundChoriocarcinoma is an aggressive malignancy of trophoblastic tissue, typically of gestational etiology. Sporadic, nongestational cases are rarely found outside of the gonads. There are only 31 cases of primary choriocarcinoma of the colon reported in the literature. As a consequence of their rarity and aggressive nature, timely diagnosis and effective treatment have proved challenging, and prognosis is very poor. For that reason, we present a rare case with prolonged survival in the youngest reported patient .Case presentationA 26-year-old Caucasian woman presented with abdominal cramping and rectal and vaginal bleeding. Elevated serum human chorionic gonadotropin and an 8-cm right-sided mass seen on ultrasound suggested ectopic pregnancy. The patient was treated with methotrexate; however, her symptoms persisted, and her human chorionic gonadotropin levels continued to rise. Further workup showed a large mass of the sigmoid colon with multiple hepatic lesions suggestive of metastases. Preliminary pathology showed adenocarcinoma. Despite surgical resection and initiation of FOLFOX chemotherapy (folinic acid, fluorouracil, oxaliplatin), the patient had significant clinical deterioration, and her human chorionic gonadotropin increased exponentially. Further pathological review showed two distinct phenotypes: adenocarcinoma merging with choriocarcinoma. The result of evaluation of the metastatic lesions was also positive for choriocarcinoma. Treatment was promptly changed to a choriocarcinoma-targeting chemotherapy regimen of EMA/CO (etoposide, methotrexate, actinomycin D, cyclophosphamide, vincristine), resulting in rapid and dramatic response. The patient had mild progression after 1 year and was switched back to FOLFOX with bevacizumab. After five cycles, scans showed further progression, and the patient was started on third-line therapy with FOLFIRI (folinic acid, fluorouracil, irinotecan) and bevacizumab. Eighteen months after her diagnosis, the patient was alive and maintaining an overall response.ConclusionsOur patient achieved a marked response and prolonged survival. Although a comprehensive review of the literature showed that survival with these tumors has improved over the past 10 years, prognosis remains poor. Currently, there is no established algorithm for the management of these rare tumors, but both the literature and our patient’s case indicate that a choriocarcinoma-targeted regimen is critical for survival. Further evaluation of these rare tumors is warranted in order to identify pathological patterns that may help in the diagnosis, management, and survival of these malignancies.

Shrub-dwelling arthropod assemblages respond differently to grazing disturbance in the southern Monte, Argentina

Grazing disturbance affects an extensive area of the Monte desert (Argentina). Arthropods are the main faunistic component of the Monte and play key roles in ecosystem processes. The aim of this study was to analyse the effect of grazing on the canopy arthropod assemblages that inhabit three representative shrub species of the southern Monte (Chuquiraga avellanedae, Schinus johnstonii and Larrea divaricata). Three sampling sites were established in three paddocks grazed by sheep and another three sites in a grazing exclosure. Five specimens of each shrub species were selected per site. Canopy arthropods were sampled using the beating method. The response of arthropod assemblages to grazing was found to vary among shrub species and between seasons. The assemblages associated with two shrub species (L. divaricata and C. avellanedae) were modified significantly by grazing. The most marked response was a general decrease in arthropod diversity in disturbed sites. The abundance of sap-sucking arthropods increased with grazing. These results could be related, in part, to differences in canopy architecture between grazed and ungrazed sites. This study contribute to understanding the ecological impacts of grazing on biological communities of the southern Monte and will aid in developing management strategies that improve regional biodiversity outcomes.

INNV-04. CONGENITAL SPINAL GLIOBLASTOMA WITH SUSTAINED RESPONSE TO TROPOMYOSIN RECEPTOR KINASE (TRK) INHIBITION: A CASE REPORT

Abstract Large scale genomic efforts have shown that a significant percentage of infant high-grade gliomas (HGG) carry targetable gene fusions. Given the rarity of congenital spinal HGGs, little is known about the molecular features of these tumors and their response to treatment. We describe a case of a patient with congenital spinal glioblastoma. A full-term female infant with symmetric IUGR was noted to have weakness with little movement of her upper extremities. MRI of the spine showed an infiltrative and hemorrhagic intraspinal cervicothoracic neoplasm with a large hemorrhagic intramedullary component. A biopsy was done and revealed a high-grade glial neoplasm with pan-TRK expression. Next generation sequencing (NGS) confirmed a MEF2D-NTRK1 gene fusion. Prior to the result of her molecular testing, the patient clinically deteriorated with decreased movement and increased apneic episodes despite steroid treatment. She was then started on treatment with carboplatin and etoposide per Macy et al 1. After one cycle of chemotherapy and given the results of NGS, targeted therapy with larotrectinib was added to her treatment. The patient showed a response (clinical and imaging) following cycle 1 with chemotherapy, and a marked response following cycle 2 with both chemotherapy and targeted TRK inhibition. The patient completed six cycles of chemotherapy and has continued on daily oral larotrectinib. She continues to tolerate the targeted therapy well and currently is 8 months old, growing and developing well. She has regained much of her movement and is eating and growing with apparent normal development. This case provides an example of a patient with congenital glioblastoma with TRK gene fusion that showed an impressive response to targeted therapy combined with more conventional chemotherapy. 1Macy M, et al. Clinical and molecular characteristics of congenital glioblastoma. Neuro Oncol. Jul;14 (7) 931–941, 2012).

PATH-44. AN UNUSUAL PRESENTATION OF A PEDIATRIC MIDLINE H3K27M-MUTANT TUMOR WITH DISSEMINATED CRANIOSPINAL LEPTOMENINGEAL DISEASE

Abstract BACKGROUND H3K27M-mutant midline lesions were recently reclassified by the WHO as “Diffuse Midline Glioma” (DMG) based on their molecular signature. DMG is one of the most common and most lethal pediatric brain tumors; terminal progression is typically caused by local progression or secondary leptomeningeal dissemination. H3K27M mutations have also been infrequently associated with a histologically diverse set of lesions, particularly spinal masses with early leptomeningeal spread, but the role of H3K27M in these atypical lesions remains poorly understood. CASE PRESENTATION A 15-year-old girl was found to have a T2/FLAIR-hyperintense and heterogeneously contrast-enhancing thalamic mass accompanied by severe leptomeningeal enhancement along the entire neuraxis. Initial infectious workup was negative, and open intracranial biopsy was inconclusive. Follow-up spinal arachnoid biopsy was diagnostic for H3K27M neuroepithelial tumor, thereafter classified as DMG. Biopsy also showed focal p53 immunopositivity, variable immunoreactivity for GFAP and synaptophysin, and an increased proliferation index, altogether suggestive of a glioneuronal origin. She received craniospinal irradiation (CSI) to 41.4Gy with a boost to the thalamic lesion to 54Gy. Imaging 1-month post-radiation demonstrated significant treatment response only with residual enhancement at the conus. CONCLUSIONS This case report describes the unique presentation of an H3K27M-mutant midline lesion with significant craniospinal leptomeningeal spread on admission and atypical glioneuronal histopathological markers. Given her avid leptomeningeal disease, spinal dural biopsy could have been considered earlier given its diagnostic yield in classifying the lesion as DMG. This lesion, however, additionally demonstrated synaptophysin positivity. Per extensive literature review, this histopathology is also potentially consistent with diffuse leptomeningeal glioneuronal tumor (DLGNT). This patient’s uniquely marked treatment response to CSI is also not typically observed in DMG, further supporting the possibility of an alternative molecular identity. In atypical DMG cases, particularly with early leptomeningeal spread, additional consideration of clinical and histopathological context is warranted for accurate diagnosis and prognostication.

Evidence of Synergistic Effect of Idasanutlin and Navitoclax Using T-Cell Acute Lymphoblastic Leukemia Patient-Derived Xenografts

T-cell acute lymphoblastic leukemia (T-ALL) is diagnosed in approximately 2,500 individuals in the United States each year. The five-year event-free survival rate for this aggressive hematologic malignancy declines with age, ranging from 70-75% for pediatric patients to 21.7% for patients aged 60-79. Thus, there is an unmet clinical need for better therapies for T-ALL. Mutations that result in Notch activation are present in the majority of T-ALL and are thought to induce transformation, in part, through activation of c-Myc. Increased expression of c-Myc induces apoptosis through a p19(ARF)-Mdm2-p53 tumor suppressor pathway. Thus, it is not surprising that over 70% of T-ALL cases have inactivating mutations in CDKN2A (p19Arf). Of note, greater than 95% of primary T-ALL cases and 75% of relapsed cases maintain wild type p53. Based on these observations, we hypothesize that treatment with MDM2 inhibitors, by bypassing the loss of ARF, will increase p53 expression and lead to T-ALL cell apoptosis. We further hypothesize that inhibition of pro-survival BH3 domain proteins, in particular BCLXL, will synergize with MDM2 inhibition to kill T-ALL cells. Of note, a prior study showed that most T-ALL (~62%) respond to treatment with ABT-263 (navitoclax), an inhibitor of BCL2, BCLXL, and BCLW, although most responses were transient. To test this hypothesis, we assessed the response of four different T-ALL xenografts to treatment with idasanutlin (a second generation MDM2 inhibitor) alone, ABT-263 (navitoclax) alone, or their combination. In each case, human T-ALL cells were injected into immunodeficient NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl) mice. Leukemic burden was measured in the blood by flow cytometry for human CD45+ cells. Once engraftment of T-ALL was observed, mice were randomized to one of four treatments: 1) ABT-263 100 mg/kg by oral gavage daily for 14 days; 2) idasanutlin 40 mg/kg by oral gavage daily on a 5-days-on 2-days-off schedule for 14 days; 3) combined idasanutlin and ABT-263; and 4) vehicle alone. Two independent experiments were performed with cohort sizes ranging from 15 (3-4 per treatment) to 31 (7-8 per treatment). Treatment with ABT-263 alone induced a significant decrease in T-ALL burden in all 4 xenotransplants (Figure 1). Treatment with idasanutlin alone induced a significant decrease in T-ALL burden in 3 of 4 xenotransplants. The non-responding xenograft was an ETP T-ALL. Finally, a marked response to combination treatment with idasanutlin and ABT-263 was seen in all four T-ALL xenografts. Indeed, the combination was found to be synergistic in each case based on a modified Bliss Independence test, analyzing average daily change for synergy interaction. In addition, overall survival was significantly increased in the combination treatment group. These data show that the combination of idasanutlin and ABT-263 is highly active in T-ALL and merits consideration for clinical testing. Disclosures Johansson: Roche: Other: Roche provided Idasanutlin free of charge. No compensation was provided, and Roche was not involved in design, conduction, or analysis of experiments.. Gauthier:Roche: Other: Roche provided Idasanutlin free of charge. No compensation was provided, and Roche was not involved in design, conduction, or analysis of experiments.. Link:Roche: Other: Roche provided Idasanutlin free of charge. No compensation was provided, and Roche was not involved in design, conduction, or analysis of experiments..

Stress-Related Responses to Alternations between Repetitive Physical Work and Cognitive Tasks of Different Difficulties.

Alternating between physical and cognitive tasks has been proposed as an alternative in job rotation, allowing workers to recover from the physical work while still being productive. However, effects of such alternations on stress have not been investigated. This controlled experiment aimed at determining the extent to which stress-related responses develop during alternating physical and cognitive work, and to determine the extent to which cognitive task (CT) difficulty influences these responses. Fifteen women performed three sessions of 10 consecutive work bouts each including a seven-minute repetitive physical task (pipetting) and a three-minute CT (n-back) at one of three difficulty levels. Stress was assessed in terms of changes in heart rate variability, blood pressure, salivary alpha-amylase, salivary cortisol, perceived stress, and cognitive performance. The work session did not result in any marked stress response, and CT difficulty did not significantly influence stress, apart from alpha-amylase being higher at the easiest CT (F = 5.34, p = 0.02). Thus, according to our results, alternating between repetitive physical tasks and cognitive tasks may be a feasible alternative to classic job rotation between physical tasks only, even if the cognitive task is quite difficult. Future studies should address possible effects of the temporal pattern of alternations, and combine even other occupationally relevant tasks, preferably for extended periods of time.

Reduction in intestinal colonization and invasion of internal organs after challenge by homologous and heterologous serovars of Salmonella enterica following vaccination of chickens with a novel trivalent inactivated Salmonella vaccine

ABSTRACT A novel inactivated vaccine, comprising three serovars of Salmonella enterica (Enteritidis, serogroup O:9; Typhimurium, serogroup O:4; Infantis, serogroup O:7) grown under conditions of iron restriction and adjuvanted with aluminium hydroxide, was evaluated for efficacy following challenge by homologous and heterologous serovars. Chickens were vaccinated at 6 and 10 weeks of age by the intramuscular route and challenged 4 to 9 weeks after the second vaccination with serovars belonging to serogroup O:9 (Enteritidis), O:4 (Typhimurium and Heidelberg), O:7 (Infantis and Virchow), and O:8 (Hadar). All vaccinated birds produced a marked systemic antibody response against each of the component vaccine antigens by the time of challenge. Significant reductions in both colonization of the intestinal tract and invasion of internal organs were observed in vaccinated birds compared with non-vaccinated controls, irrespective of the challenge serovar. The findings suggest that broad serovar protection within the constitutive serogroups of an inactivated multi-valent vaccine is possible and could, therefore, play an important role in future Salmonella control programmes. RESEARCH HIGHLIGHTS Novel inactivated trivalent Salmonella chicken vaccine was developed and tested. Vaccine induced marked systemic antibody response against all vaccine antigens. Significant reductions in intestinal tract colonization and internal organ invasion. Vaccine efficacy demonstrated against homologous and heterologous serovars.

Advanced glycation end-products disrupt brain microvascular endothelial cell barrier: The role of mitochondria and oxidative stress

During diabetes mellitus, advanced glycation end-products (AGEs) are major contributors to the development of alterations in cerebral capillaries, leading to the disruption of the blood-brain barrier (BBB). Consequently, this is often associated with an amplified oxidative stress response in microvascular endothelial cells. As a model to mimic brain microvasculature, the bEnd.3 endothelial cell line was used to investigate cell barrier function. Cells were exposed to native bovine serum albumin (BSA) or modified BSA (BSA-AGEs). In the presence or absence of the antioxidant compound, N-acetyl-cysteine, cell permeability was assessed by FITC-dextran exclusion, intracellular free radical formation was monitored with H2DCF-DA probe, and mitochondrial respiratory and redox parameters were analyzed. We report that, in the absence of alterations in cell viability, BSA-AGEs contribute to an increase in endothelial cell barrier permeability and a marked and prolonged oxidative stress response. Decreased mitochondrial oxygen consumption was associated with these alterations and may contribute to reactive oxygen species production. These results suggest the need for further research to explore therapeutic interventions to restore mitochondrial functionality in microvascular endothelial cells to improve brain homeostasis in pathological complications associated with glycation.