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Raw milk is considered a high-risk source of Campylobacter due to faecal contamination from healthy cattle and farm environments, thus linking raw milk consumption to global outbreaks. Detection of Campylobacter in raw milk poses challenges due to low contamination levels and antibacterial properties of the milk. Culture-based protocols for Campylobacter detection in milk vary, mainly with regard to pH adjustment and the choice of enrichment broth. This European collaborative study was organised by the EU Reference Laboratory (EURL) for Campylobacter together with eight EU National Reference Laboratories (NRL) for Campylobacter with the purpose to evaluate methods for culture-based detection of Campylobacter in raw cow's milk. The study was divided into two parts, an interlaboratory part and an intralaboratory part, both organised around the same two protocols. The aim of protocol 1 was to evaluate the impact of pH adjustment and storage of the milk on the culturability of Campylobacter over time. Aliquots of the spiked milk were adjusted either to pH 7.0 or pH 7.6 or left unadjusted. The milk was stored up to 48 h at refrigerated temperature and Campylobacter was quantified according to ISO 10272-2 on day 0, 1 and 2. The aim of protocol 2 was to evaluate which enrichment broth, Bolton broth (BB) or Preston broth (PB), showed highest sensitivity in detection of Campylobacter. The spiked milk was enriched in BB and PB as described in ISO 10272-1:2017 or ISO 10272-1:2017/Amd1.2023. In the interlaboratory part, each milk batch was collected locally by each participating NRL/EURL and inoculated with the same Campylobacter strain. In the follow-up intralaboratory part, the EURL-Campylobacter repeated the tests in protocol 1 and 2 but used different Campylobacter strains and strains subjected to thermal stress prior to inoculation. The results show that pH adjustment of raw milk has a negligible impact on culture-based detection of Campylobacter, regardless of strain and level of environmental stress. The composition of milk and properties of the inoculated strain influence culture-based detection of Campylobacter over storage time, and strains subjected to additional stress prior to inoculation in milk are reduced in culturability much faster than the same strains prepared under normal conditions. Finally, the study showed that PB without Campylobacter growth supplement is less effective than BB in detecting Campylobacter in raw milk., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Diffuse hemispheric glioma, H3G34R/V-mutant (DHG-H3G34) is characterized by poor prognosis and lack of effective treatment options. DHG-H3G34R further harbor deactivation of Alpha-Thalassemia/Mental Retardation Syndrome X-linked protein (ATRX; DHG-H3G34R_ATRX) suggesting a unique interaction of these two oncogenic alterations. In this study, we dissect their cell biological interplay, investigate the impact on telomere stabilization and, consequently, validate a targeted therapy approach., Methods: We characterized patient-derived primary pediatric high-grade glioma (pHGG) models for telomere-maintenance mechanisms, DNA damage stress (including protein expression, pH2AX/Rad51 foci, cell-cycle arrest) and their sensitivity towards poly-ADP polymerase inhibitor (PARPi) combinations. Human induced pluripotent stem cells (iPSCs) were used for modelling the disease. The anticancer activity of PARPi combinations in vivo was studied in Chorioallantoic Membrane (CAM) and orthotopic in vivo experiments. Finally, we treated a DHG-H3G34R_ATRX patient with a PARPi combination therapy., Results: We elaborate that alternative lengthening of telomeres (ALT) is a key characteristic of DHG-H3G34R_ATRX. A dominant cooperative effect between H3G34R and ATRX loss in ALT activation also became apparent in iPSCs, which endogenously exert telomerase activity. In both, patient-derived DHG-H3G34R_ATRX models and H3G34R+/ATRX- iPSCs, the ALT phenotype was associated with increased basal DNA damage stress, mediating synergistic susceptibility towards PARPi (talazoparib, niraparib) combinations with topoisomerase-I inhibitors (topotecan, irinotecan). In a first-of-its-kind case, treatment of a DHG-H3G34R_ATRX patient with the brain-penetrant PARP inhibitor niraparib and topotecan resulted in a significant tumor reduction., Conclusion: Our preclinical and clinical data strongly support the further development of PARPis together with DNA damage stress-inducing treatment regimens for DHG-H3G34R_ATRX., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Neuro-Oncology.)

Introduction: There have been two cases of congenital diaphragmatic hernia associated with a gastroesophageal duplication cyst documented in the literature, both presented symptomatically in the newborn period. This case is unique given the patient presented in adolescence asymptomatically., Presentation of Case: We present a case of a 15-year-old female who initially presented with lower abdominal pain. A CT chest and abdomen was concerning for a moderate to large hiatal vs diaphragmatic hernia and an adjacent 4.3 cm lower mediastinal cyst. Though asymptomatic, the patient underwent diagnostic thoracoscopy with cyst excision. On post operative day one, the patient endorsed severe uncontrolled abdominal pain with multiple bouts of non-bilious emesis. Chest X-ray and upper GI were positive for herniation of the stomach through the diaphragm. The patient was taken back to the operating room emergently for diagnostic thoracoscopy where the stomach was found herniated through a diaphragmatic defect with no evidence of necrosis. The stomach was reduced into the abdominal cavity. The patient tolerated this procedure well, recovered appropriately and was seen two-weeks post-operatively fully recovered., Discussion: Better visualization and thorough tactile check of the diaphragmatic defect during the initial operation would increase the chance of detecting a defect in the diaphragm immediately and avoid subsequent herniation of abdominal contents., Conclusion: When Foregut duplication cysts exist near the diaphragm it is critical to get a thorough view of the anatomy and ensure no diaphragmatic defect exits., Competing Interests: Conflict of interest statement The authors have no conflict of interests to declare., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Liquid biopsy diagnostic methods are an emerging complementary tool to imaging and pathology techniques across various cancer types. However, there is still no established method for the detection of molecular alterations and disease monitoring in MB, the most common malignant CNS tumor in the pediatric population. In the presented study, we investigated droplet digital polymerase chain reaction (ddPCR) as a highly sensitive method for the detection of MYC amplification in bodily fluids of group 3 MB patients., Methods: We identified a cohort of five MYC -amplified MBs by methylation array and FISH. Predesigned and wet-lab validated probes for ddPCR were used to establish the detection method and were validated in two MYC -amplified MB cell lines as well as tumor tissue of the MYC -amplified cohort. Finally, a total of 49 longitudinal CSF samples were analyzed at multiple timepoints during the course of the disease., Results: Detection of MYC amplification by ddPCR in CSF showed a sensitivity and specificity of 90% and 100%, respectively. We observed a steep increase in amplification rate (AR) at disease progression in 3/5 cases. ddPCR was proven to be more sensitive than cytology for the detection of residual disease. In contrast to CSF, MYC amplification was not detectable by ddPCR in blood samples., Conclusions: ddPCR proves to be a sensitive and specific method for the detection of MYC amplification in the CSF of MB patients. These results warrant implementation of liquid biopsy in future prospective clinical trials to validate the potential for improved diagnosis, disease staging and monitoring.

Objective: The intraoperative visualization of adult-type diffuse gliomas with 5-aminolevulinic acid (5-ALA) induced fluorescence is widely used in the neurosurgical field. While visible 5-ALA induced fluorescence is found in the majority of high-grade gliomas, most low-grade gliomas lack visible fluorescence during surgery. Recently, the heme biosynthesis pathway was identified as crucial influencing factor for presence of visible fluorescence since it metabolizes 5-ALA to fluorescing Protoporphyrin IX (PpIX). However, the exact alterations within the heme biosynthesis pathway resulting in visible 5-ALA induced fluorescence in gliomas are still unclear. The aim of the present study was thus to compare the mRNA and protein expression of promising intramitochondrial heme biosynthesis enzymes/transporters in glioma tissue samples of different fluorescence behavior., Methods: A total of 19 strongly fluorescing and 21 non-fluorescing tissue samples from neurosurgical adult-type diffuse gliomas (WHO grades II-IV) were included in the current analysis. In these samples, we investigated the mRNA expression by quantitative real time PCR and protein expression using immunohistochemistry of the intramitochondrial heme biosynthesis enzymes Coproporphyrinogen Oxidase (CPOX), Protoporphyrinogen Oxidase (PPOX), Ferrochelatase (FECH), and the transporter ATP-binding Cassette Subfamily B Member 2 (ABCG2)., Results: Regarding mRNA expression analysis, we found a significantly decreased ABCG2 expression in fluorescing specimens compared to non-fluorescing samples ( p = 0.001), whereas no difference in CPOX, PPOX and FECH was present. With respect to protein expression, significantly higher levels of CPOX ( p = 0.005), PPOX ( p < 0.01) and FECH ( p = 0.003) were detected in fluorescing samples. Similar to mRNA expression analysis, the protein expression of ABCG2 ( p = 0.001) was significantly lower in fluorescing samples., Conclusion: Distinct alterations of the analyzed heme biosynthesis factors were found primarily on protein level. Our data indicate that heme biosynthesis pathway activity in general is enhanced in fluorescing gliomas with upregulation of PpIX generating enzymes and decreased ABCG2 mediated PpIX efflux outweighing the also increased further metabolization of PpIX to heme. Intramitochondrial heme biosynthesis factors thus constitute promising pharmacological targets to optimize intraoperative 5-ALA fluorescence visualization of usually non-fluorescing tumors such as low-grade gliomas., Competing Interests: AB had research support from Daiichi Sankyo and honoraria for lectures, consultation or advisory board participation from Roche Bristol-Meyers Squibb, Merck, Daiichi Sankyo as well as travel support from Roche, Amgen, and AbbVie. GW had received restricted travel support from NX Development Corp. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Mischkulnig, Roetzer-Pejrimovsky, Lötsch-Gojo, Kastner, Bruckner, Prihoda, Lang, Martinez-Moreno, Furtner, Berghoff, Woehrer, Berger, Widhalm and Kiesel.)

The worldwide prevalence of dementia is estimated at 35.6 million and will rise to 115 million by 2050. There is therefore an urgent need for well-founded dementia diagnostics and well-researched therapeutic options. Previous studies have highlighted that spermidine has the ability to trigger the important process of dissolving amyloid-beta plaques by autophagy. They also confirmed that nutritional intervention with the natural polyamine spermidine can prevent memory loss in aging model organisms. This multicentric double-blind preliminary study focused on the effect of oral spermidine supplementation on older adults' cognitive performance. Memory tests were carried out on 85 subjects aged between 60 and 96 years in 6 nursing homes in Styria. Blood samples were taken for the determination of spermidine concentration and measurement of metabolic parameters. The results demonstrated a clear correlation between the intake of spermidine and the improvement in cognitive performance in subjects with mild and moderate dementia in the group treated with the higher spermidine dosage. The most substantial improvement in test performance was found in the group of subjects with mild dementia with an increase of 2.23 points (p = 0.026) in the mini mental state examination (MMSE) and 1.99 (p = 0.47) in phonematic fluidity. By comparison, the group which had a lower spermidine intake showed consistent or declining cognitive performance.

Background: Renal lymphoma in dogs is rare and has a poor prognosis. Granular lymphocyte morphology is rarely reported in canine renal lymphoma. Mild to moderate polycythemia is reported in a number of canine renal lymphoma cases., Case Presentation: A 10-year-old Labrador retriever presented to a university veterinary teaching hospital after a 1-month history of polyuria, polydipsia, and pollakiuria and a 2-week history of abdominal distention, lethargy, and increased respiratory effort. Abdominal ultrasound showed a wedge-shaped to rounded, heterogeneously hypoechoic mass lesion in the left kidney. Cytologic analysis of a percutaneous aspirate of the mass was consistent with lymphoma of granular lymphocytes. Severe polycythemia (hematocrit 0.871) was noted on a complete blood cell count. Clonality analysis identified a clonally rearranged T-cell receptor (TCR) gene and immunohistochemical staining was CD3+, CD79a- and CD11d+, supporting cytotoxic T-cell lymphoma., Conclusions: To our knowledge, this is the first report of renal cytotoxic T-cell lymphoma with severe polycythemia in a dog. Severe polycythemia and renal cytotoxic T-cell lymphoma are both rare in dogs; this report adds to the body of knowledge on these conditions.

Objective: To characterize objective and subjective elements of the personal lives of American College of Veterinary Surgeons (ACVS) diplomates and identify elements of personal life associated with professional life or career success., Sample: 836 ACVS diplomates., Procedures: An 81-item questionnaire was sent to 1,450 diplomates in 2015 via email and conducted by means of an online platform. Responses were analyzed to summarize trends and identify associations among selected variables., Results: Men were more likely than women to be married or in a domestic partnership (88% vs 68%, respectively) and to have children (77% vs 47%). Among women but not men, respondents in large animal practice were less likely than were those in small animal practice to be represented in these categories. Women had children later in their career than did men and indicated that their stage of training played an important role in family planning. Respondents with children worked significantly fewer hours than did those without children, with a greater reduction in hours for women versus men (6.0 vs 3.1 hours, respectively). Women were more likely to require external childcare services than were men. Women were more likely to report that having children had negatively impacted their professional lives. No negative associations between measures of professional success (eg, advancement or personal income) and parenthood were identified., Conclusions and Clinical Relevance: Family demographics differed between male and female ACVS diplomates, yet no objective impact on career success was identified. Work-life balance may play an important role in recruitment, retention, and job satisfaction of veterinary surgeons.

Objective: To characterize elements of employment, professional success, and personal life for American College of Veterinary Surgeons (ACVS) diplomates and identify elements of practice that may serve as barriers to work-life balance or affect men and women differently., Sample: 836 ACVS diplomates., Procedures: An 81-item questionnaire was sent to 1,450 ACVS diplomates in 2015 via email and conducted by means of an online platform. Responses were analyzed to identify associations among selected variables., Results: The survey response rate was 58% (836/1,450). The median age category among respondents was 41 to 45 years. The ratio of male to female diplomates was equivalent among those < 40 years old. Respondents in small animal private practice worked the fewest number of hours; those in equine or large animal private practice worked the most and had the most on-call responsibility. Women were more likely than men to be employed in academia. In both private practice and academia, respondents in small animal practice earned more than did those in equine or large animal practice, and women earned less than did men, even after adjustment for relevant covariates. Women were less likely than men to be practice owners or to hold a prestigious academic title and rank. Perceptions about the effect of gender in the workplace differed between men and women., Conclusions and Clinical Relevance: Findings suggested that the veterinary surgical profession is demanding for both genders, although increased flexibility in certain areas may improve work-life balance.