[RCVS: case-study and role of substance abuse, Covid and psychotropic drugs]. / Reversibel cerebraal vasoconstrictie­syndroom: casus en rol middelengebruik, COVID-19 en psychofarmaca.

Reversible cerebral vasoconstriction syndrome (RCVS) is a condition defined by severe sudden-onset headaches, typically ‘thunderclap’ headaches, caused by multifocal cerebral vasoconstriction. Various triggers have been described, including illegal substances, medication and infections. We observed a 27 year old man that suddenly developed severe headaches during admission to a psychiatric ward, where RCVS was diagnosed as most likely clinical cause. He was given nimodipine with rapid and full symptom remission. We aim to highlight this rare, but important, neurological syndrome and its various psychiatric risk factors.

Quality of life in patients and caregivers after aneurysmal subarachnoid hemorrhage: a Flemish population study.

BACKGROUND AND AIMS: Aneurysmal subarachnoid hemorrhages (aSAH) have high mortality and morbidity. However, the impact on Quality of Life (QoL) of patients remains poorly documented, and data on primary caregiver burden is even scarcer. METHODS: This is a single center, cross-sectional study performed at the Antwerp University Hospital, Belgium. We included aSAH patients during follow-up at the outpatient clinic and assessed the QoL, by using the Stroke Specific Quality of Life scale (SSQoL). Caregiver burden was evaluated by the Caregiver Strain Index (CSI). The aSAH severity and functional outcome (at 90 days) were assessed, respectively, by mFisher score and modified Ranking Scale (mRS). Statistical analysis was performed using SPSS version 27. RESULTS: In total, 22 aSAH patients were included, on average 15.5 (range 4-45) months after the aSAH. The SSQoL score was 3.7 ± 0.7, with a mean psychosocial domain score of 3.2 ± 0.8 and physical domain of 4.2 ± 0.8. Psychosocial factors, especially decreased energy levels and cognitive impairment, had a negative impact on the QoL (p = 0.02 en p = 0.05). No association was found between QoL and mFisher, nor between QoL and mRS. Fifteen primary caregivers completed the CSI. Only 3 (20%) of them reported a high care burden (CSI > 6), although changes in daily life and personal plans were reported, respectively, by 73% (n = 11) and 67% (n = 10) of caregivers. We only found a correlation between the mFisher score and CSI (p = 0.01). CONCLUSION: Our results emphasize that there is an important psychosocial impact on the QoL of patients after aSAH, and their primary caregivers. More research is warranted.

A high protein meal affects plasma insulin concentrations and amino acid metabolism in horses with equine metabolic syndrome.

Equine metabolic syndrome (EMS) is characterized by an abnormal insulin response to a glycemic challenge but despite the known insulinotropic effects of certain amino acids, there is a paucity of data evaluating the impact of dietary protein on insulin dynamics in these horses. The objective was therefore to assess insulin and amino acid responses following intake of a high protein meal in healthy horses and those with EMS. Six mature horses diagnosed with EMS and six age-matched control horses without EMS were used. Horses were fed 2g/kg body mass (BM) of a high protein pellet (31% crude protein) at time 0 and 30min, for a total of 4g/kg BM, following an overnight fast. Blood samples collected during a 4h period were analysed for plasma glucose, insulin, amino acids and urea concentrations. Glucose concentrations were not different between groups (P=0.2). Horses with EMS had a 9-fold greater insulinemic response to the consumption of a high protein meal compared with controls (P=0.046). Post-prandial levels of histidine, citrulline, tyrosine, valine, methionine, isoleucine, leucine and ornithine were higher in horses with EMS (P<0.05). Baseline urea nitrogen concentrations were not significantly different between groups (P=0.1). Knowing that certain amino acids are insulin secretagogues, these results illustrate that consumption of a high protein meal caused a hyperinsulinemic response and affected amino acid dynamics in horses with EMS. These findings suggest that dietary protein content should be taken into consideration in the management of horses with insulin dysregulation.

A chlorhexidine-releasing epoxy-based coating on titanium implants prevents Staphylococcus aureus experimental biomaterial-associated infection.

Prevention of biomaterial-associated infections (BAI) remains a challenging problem, in particular due to the increased risk of resistance development with the current antibiotic-based strategies. Metallic orthopaedic devices, such as non-cemented implants, are often inserted under high mechanical stress. These non-cemented implants cannot be protected by e.g. antibioticreleasing bone cement or other antimicrobial approaches, such as the use of bioactive glass. Therefore, in order to avoid abrasion during implantation procedures, we developed an antimicrobial coating with great mechanical stability for orthopaedic implants, to prevent Staphylococcus aureus BAI. We incorporated 5 and 10 wt % chlorhexidine in a novel mechanically stable epoxy-based coating, designated CHX5 and CHX10, respectively. The coatings displayed potent bactericidal activity in vitro against S. aureus, with over 80 % of the release (19 µg/cm2 for CHX5 and 41 µg/cm2 for CHX10) occurring within the first 24 h. In mice, the CHX10 coating significantly reduced the number of CFU (colony forming units), both on the implants and in the peri-implant tissues, 1 d after S. aureus challenge. The CHX10-coated implants were well-tolerated by the animals, with no signs of toxicity observed by histological analysis. Moreover, the coating significantly reduced the frequency of culture-positive tissues 1 d, and of culture-positive implants 1 and 4 d after challenge. In summary, the chlorhexidine-releasing mechanically stable epoxy-based CHX10 coating prevented implant colonisation and S. aureus BAI in mice and has good prospects for clinical development.

Characteristics of Queensland physicians and the influence of rural exposure on practice location.

The Queensland branch of the Royal Australasian College of Physicians (RACP) commissioned this study to update their workforce profile and examine rural practice. The present investigation aimed to describe characteristics of Queensland physicians and determine the influence of childhood and training locations on current rural practice. A cross-sectional online survey, conducted 4 July-4 November 2013, was administered to Fellows of The RACP, Queensland. Descriptive statistics report characteristics and logistic regression analyses identify associations and interactions. The outcome measure was current practice location using the Australian Standard Geographic Classification - Remoteness Area. Data were obtained for 633 physicians. Their average age was 49.5 years, a third was female and a quarter was in rural practice. Rural practice was associated with a rural childhood (odds ratio (OR) (95% confidence interval, CI) 1.89 (1.10, 3.27) P = 0.02) and any time spent as an intern (OR 4.07 (2.12, 7.82) P < 0.001) or registrar (OR 4.00 (2.21, 7.26) P < 0.001) in a rural location. Physicians with a rural childhood and rural training were most likely to be in rural practice. However, those who had a metropolitan childhood and a rural internship were approximately five times more likely to be working in rural practice than physicians with no rural exposure (OR 5.33 (1.61, 17.60) P < 0.01). The findings demonstrate the positive effect of rural vocational training on rural practice. A prospective study would determine if recent changes to the Basic Physician Training Pathway and the Basic Paediatric Training Network (more rural training than previous pathways) increases the rate of rural practice.

Therapeutic expression of hairpins targeting apolipoprotein B100 induces phenotypic and transcriptome changes in murine liver.

Constitutive expression of short hairpin RNAs (shRNAs) may cause cellular toxicity in vivo and using microRNA (miRNA) scaffolds can circumvent this problem. Previously, we have shown that embedding small interfering RNA sequences targeting apolipoprotein B100 (ApoB) in shRNA (shApoB) or miRNA (miApoB) scaffolds resulted in differential processing and long-term efficacy in vivo. Here we show that adeno-associated virus (AAV)-shApoB- or AAV-miApoB-mediated ApoB knockdown induced differential liver morphology and transcriptome expression changes. Our analyses indicate that ApoB knockdown with both shApoB and miApoB resulted in alterations of genes involved in lipid metabolism. In addition, in AAV-shApoB-injected animals, genes involved in immune system activation or cell growth and death were affected, which was associated with increased hepatocyte proliferation. Subsequently, in AAV-miApoB-injected animals, changes of genes involved in oxidoreductase activity, oxidative phosphorylation and nucleic bases biosynthetic processes were observed. Our results demonstrate that long-term knockdown of ApoB in vivo by shApoB or miApoB induces several transcriptome changes in murine liver. The increased hepatocyte profileration by AAV-shRNA may have severe long-term effects indicating that AAV-mediated RNA interference therapy using artificial miRNA may be a safer approach for familial hypercholesterolemia therapy.

Intraplaque hemorrhage in cardiac allograft vasculopathy.

Plaque hemorrhage, inflammation and microvessel density are key determinants of plaque vulnerability in native coronary atherosclerosis (ATS). This study investigates the role of intraplaque hemorrhage (IPH) and its relation with inflammation and microvessels in cardiac allograft vasculopathy (CAV) in posttransplanted patients. Seventy coronary plaques were obtained from 12 patients who died because of CAV. For each patient we collected both native heart and the allograft, at the time of transplantation and autopsy, respectively. Intralesion inflammation, microvessels and IPH were assessed semi-quantitatively. IPH was observed in 21/35 (60%) CAV lesions and in 8/35 (22.9%) native ATS plaques, with a strong association between fibrocellular lesions and IPH (p = 0.0142). Microvessels were detected in 26/35 (74.3%) of CAV lesions with perivascular leakage as sign of endothelial damage in 18/26 (69.2%). IPH was strongly associated with microvessels (p < 0.0001). Inflammation was present in 31/35 (88.6%) of CAV lesions. CAV IPH+ lesions were characterized by presence of both fresh and old hemorrhage in 12/21 (57.1%). IPH, associated with microvessel damage and inflammation, is an important feature of CAV. Fresh and old intralesion hemorrhage suggests ongoing remodeling processes promoting the lesion progression and vulnerability.

Investigation of the performance of digital mammographic X-ray equipment: determination of noise equivalent quanta (NEQQC) and detective quantum efficiency (DQEQC) compared with the automated analysis of CDMAM test images with CDCOM and CDIC programs.

PURPOSE: The purpose of this study was to determine the values for noise equivalent quanta, detective quantum efficiency, modulation transfer function, noise power spectrum, and the values for the parameters for automated CDMAM test phantom analyses required to achieve satisfactory quality of digital mammograms. MATERIALS AND METHODS: During the course of tests according to PAS 1054 (8 CR and 12 DR systems), test images were made with a test phantom insertion plate containing two lead edges in nearly horizontal and vertical directions. Only original data were processed with a program that was developed at the Cologne University of Applied Sciences (FH-Köln). All equipment systems complied with the requirements regarding visual recognition of gold-plated mammo detail test objects. CDMAM test images were also evaluated using the CDIC (CUAS) and CDCOM (EUREF) programs. RESULTS: CDMAM test images show comparable values for the parameters, precision, sensitivity and specificity. DR systems require about half the dose used for CR systems for similar results. The NEQ values achieved with the dose used for the CDMAM test images show larger scatter ranges. The MTF of the different equipment system types differ significantly from each other. CONCLUSION: Visual evaluation of CDMAM test images can be replaced by automated evaluation. Limiting values were determined for each parameter. Automated evaluation of CDMAM test phantom images should be used to determine the physical parameter NEQQC. This method is much more sensitive to noise and sharpness influences and has a higher validity than diagnostic methods. Automated evaluation objectivizes testing.

The TNF family member APRIL promotes colorectal tumorigenesis.

The tumor necrosis factor (TNF) family member APRIL (A proliferation inducing ligand) is a disease promoter in B-cell malignancies. APRIL has also been associated with a wide range of solid malignancies, including colorectal cancer (CRC). As evidence for a supportive role of APRIL in solid tumor formation was still lacking, we studied the involvement of APRIL in CRC. We observed that ectopic APRIL expression exacerbates the number and size of adenomas in Apc(Min) mice and in a mouse model for colitis-associated colon carcinogenesis. Furthermore, knockdown of APRIL in primary spheroid cultures of colon cancer cells and both mouse and human CRC cell lines reduced tumor clonogenicity and in vivo outgrowth. Taken together, our data therefore indicate that both tumor-derived APRIL and APRIL produced by non-tumor cells is supportive in colorectal tumorigenesis.

Endothelial insulin receptor expression in human atherosclerotic plaques: linking micro- and macrovascular disease in diabetes?

OBJECTIVE: Exogenous insulin use in patients with type 2 diabetes (DM2) has been associated with an increased risk of cardiovascular events. Through which mechanisms insulin may increase atherosclerotic plaque vulnerability is currently unclear. Because insulin has been suggested to promote angiogenesis in diabetic retinopathy and tumors, we hypothesized that insulin enhances intra-plaque angiogenesis. METHODS: An in vitro model of pathological angiogenesis was used to assess the potential of insulin to enhance capillary-like tube formation of human microvascular endothelial cells (hMVEC) into a three dimensional fibrin matrix. In addition, insulin receptor expression within atherosclerotic plaques was visualized in carotid endarterectomy specimens of 20 patients with carotid artery stenosis, using immunohistochemical techniques. Furthermore, microvessel density within atherosclerotic plaques was compared between 68 DM2 patients who received insulin therapy and 97 DM2 patients who had been treated with oral glucose lowering agents only. RESULTS: Insulin, at a concentration of 10(-8)M, increased capillary-like tube formation of hMVEC 1.7-fold (p<0.01). Within human atherosclerotic plaques, we observed a specific distribution pattern for the insulin receptor: insulin receptor expression was consistently higher on the endothelial lining of small nascent microvessels compared to more mature microvessels. There was a trend towards an increased microvessel density by 20% in atherosclerotic plaques derived from patients using insulin compared to plaques derived from patients using oral glucose lowering agents only (p=0.05). CONCLUSION: Exogenous insulin use in DM2 patients may contribute to increased plaque vulnerability by stimulating local angiogenesis within atherosclerotic plaques.

Early onset of endothelial cell proliferation in coronary thrombi of patients with an acute myocardial infarction: implications for plaque healing.

AIMS: Coronary thrombotic occlusion in ST-segment elevation myocardial infarction (STEMI) patients is often preceded by episodes of progressive growth of the thrombus mass. Similar to wound healing, the organization of thrombus could depend on ingrowth of microvessels in order to stabilize its structure. We investigated the patterns of neovascularization in different stages of coronary thrombus evolution. MATERIAL AND METHODS: Thrombectomy materials obtained from STEMI patients were histologically classified according to thrombus age in three groups: fresh (< 1 day), lytic (1-5 days) or organized (> 5 days) thrombi. Forty thrombi of each group were randomly collected. Neovascularization in the thrombi was evaluated histomorphologically and with immunodouble stains to visualize various differentiation antigens of endothelial cells (ECs) and primitive cells. RESULTS: Morphologically, ECs in the coronary thrombi manifested as: single cells, cell clusters or microvessels. CD31+/CD34+ ECs were present in 98% of all the thrombi. In addition, endothelial clusters were found in 63% of the fresh thrombi (< 1 day). CD105+, Ki67+, or C-kit+ ECs (active, proliferating cells) were observed in all the stages, but significantly more in organized thrombi (> 5 days) compared with fresh and lytic ones (< 5 days), and mainly as cell clusters (P ≤ 0.05 for all). CD133+ primitive cells were found only sporadically in 11% of all the samples. CONCLUSION: EC proliferation is initiated very early, and gradually progresses during the organization process of thrombus after coronary plaque disruption, with only a limited contribution of primitive cells in this process.

[A retrospective efficacy and safety study of UVB-TL01 phototherapy and PUVA therapy in palmoplantar psoriasis]. / Étude rétrospective de l'efficacité et de la tolérance de la photothérapie UVBTL01 et de la PUVAthérapie dans le psoriasis palmoplantaire.

BACKGROUND: Topical therapy is generally insufficient in palmoplantar psoriasis. UVBTL01 phototherapy is a therapeutic alternative and we conducted a retrospective study of the efficacy and safety of this approach and of PUVA therapy in palmoplantar psoriasis. PATIENTS AND METHODS: All patients treated with UVBTL01 or PUVA therapy from November 2001 to April 2008 were included in the study. Phototherapy was given three times a week. Evaluation was performed after 20 sessions, again after 30 sessions and then at the end of the treatment. Therapeutic outcome was classed as "failure", "slight improvement" or "improvement or clear skin". RESULTS: UVBTL01 phototherapy and PUVA therapy were effective, with "improvement or clear skin" in respectively 52% and 61% of cases and "slight improvement" in 16% and 23% of cases at the end of the treatment. With UVBTL01, adverse effects occurred in 20% of cases (erythema 18%, first-degree burns 7%) and treatment was discontinued as a result in only 4% of cases. Adverse effects occurred in 50% in patients on PUVA therapy, mainly due to methoxypsoralen intake. CONCLUSION: UVBTL01 phototherapy and PUVA therapy are efficacious treatments in palmoplantar psoriasis; UVBTL01 phototherapy involves fewer constraints and has fewer adverse effects.

Could recombinant insulin compounds contribute to adenocarcinoma progression by stimulating local angiogenesis?

AIMS/HYPOTHESIS: Negative effects on the progression of adenocarcinomas by hyperinsulinaemia and the insulin analogue glargine (A21Gly,B31Arg,B32Arg human insulin) have recently been suggested. Most actions of this insulin analogue have hitherto been explained by direct stimulation of growth potential of neoplastic cells and by its IGF-1 related properties. However, insulin-stimulated angiogenesis could be an additional factor involved in tumour progression and clinical outcomes associated with cancer. METHODS: Five types of human adenocarcinoma (breast, colon, pancreas, lung and kidney) were evaluated for the presence of insulin receptors (IRs) on angiogenic structures. In an in vitro angiogenesis assay, various commercially available insulin compounds were evaluated for their potential to increase capillary-like tube formation of human microvascular endothelial cells (hMVEC). Insulin compounds used were: human insulin, insulin lispro (B28Lys,B29Pro human insulin), insulin glargine and insulin detemir (B29Lys[e-tetradecanoyl],desB30 human insulin). RESULTS: Insulin receptors were found to be strongly expressed on the endothelium of microvessels in all evaluated adenocarcinomas, in addition to variable expression on tumour cells. Low or no detectable expression of IRs was seen on microvessels in extratumoral stroma. Incubation with commercially available insulin compounds increased capillary-like tube formation of hMVEC in vitro. CONCLUSIONS/INTERPRETATION: Our results suggest that all tested insulin compounds may stimulate tumour growth by enhancing local angiogenesis. Future studies need to confirm the association between insulin therapy in type 2 diabetes and tumour progression.

[Comparison of two automatic evaluation methods on Images of the CDMAM test phantom]. / Vergleich zweier automatischer Verfahren zur Auswertung von CDMAM-Prüfkörperaufnahmen.

PURPOSE: To test the sensitivity of automatic methods for evaluating CDMAM test images with respect to noise. MATERIALS AND METHODS: CDMAM test images were analyzed with two computer programs. The images were made with different tube loads [mAs]. The other exposure conditions remained constant. They were analyzed with the CDCOM program, which is offered by the EUREF as a free download, and with the CDMAM Image Checker (CDIC), which was developed by the authors. RESULTS: The determination of the sensitivity in one image always delivers the same result when the same type of computer program is used. This means that the precision of both programs is sufficient. The dose sensitivity of CDIC is two times higher than the sensitivity of CDCOM. However, the required entrance dose (ESAK) for a faultless evaluation with the CDIC program is in the range of 10 mGy. The nominal sensitivity values for the CDCOM program attain a higher level. Differences in dose of more than 5 % should be detectable by both programs. CONCLUSION: Methods that dispense with visual inspections to determine the performance of X-ray units for mammography can be applied in the acceptance test or the yearly constancy tests according to the German X-ray directive ( section sign 16). The CDCOM program cannot be characterized fully because the data is not complete. Finally the detection methods are not clear. Therefore, the CDCOM program can be called a black box method, while the CDIC has to be called an open source method (general public license).

ACE mediates ventilator-induced lung injury in rats via angiotensin II but not bradykinin.

Ventilator-induced lung injury is characterised by inflammation and apoptosis, but the underlying mechanisms are poorly understood. The present study proposed a role for angiotensin-converting enzyme (ACE) via angiotensin II (Ang II) and/or bradykinin in acute lung injury. The authors assessed whether ACE and, if so, Ang II and/or bradykinin are implicated in inflammation and apoptosis by mechanical ventilation. Rats were ventilated for 4 h with low- or high-pressure amplitudes in the absence or presence of the ACE inhibitor captopril. Nonventilated animals served as controls. ACE activity, Ang II and bradykinin levels, as well as inflammatory parameters (total protein, macrophage inflammatory protein-2 and interleukin-6) were determined. Apoptosis was assessed by the number of activated caspase-3 and TUNEL (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick-end labelling)-positive cells. Bronchoalveolar lavage fluid ACE activity, levels of total protein, inflammatory parameters and the number of apoptotic cells were increased in the high-pressure amplitude group as compared with the control group. Blocking ACE activity by captopril attenuated inflammation and apoptosis in the latter group. Similar results were obtained by blocking Ang II receptors, but blocking bradykinin receptors did not attenuate the anti-inflammatory and anti-apoptotic effects of captopril. The current authors conclude that inflammation and apoptosis in ventilator-induced lung injury is, at least in part, due to angiotensin-converting enzyme-mediated angiotensin II production.

A focus on fixation.

Three fixation issues related to immunostaining are discussed here: 1) Generally, a tissue block is fixed, then embedded and sectioned (pre-fixation). The type of fixative applied, crosslinking or coagulating, has an impact on selecting an epitope retrieval method. Individual antigens have a fixation-retrieval characteristic. 2) A long fixation time, especially with crosslinking fixatives, may compromise the result of immunostaining. This negative effect varies among different antigens and can be partially restored by applying a more sensitive/efficient detection system such as tyramide amplification. 3) Sections cut from a fresh frozen tissue block usually are acetone fixed(post-fixation). This was accepted as the "gold standard" for a long time. Post-fixation, however,may have serious consequences for preservation of small peptides leaking from the cut open cells,whereas this is not the case with pre-fixed intact cells. Consequently, the concept of an acetone post-fixed cryostat tissue section as "gold standard" no longer exists and a more appropriate use of the terms immunohistochemistry and immunocytochemistry therefore seems justified. For many antibodies, it is not known whether a formalin fixed, paraffin embedded tissue specimen is appropriate. Suggestions are made for creating a positive control cell block for testing such antibodies.

Colocalisation of intraplaque C reactive protein, complement, oxidised low density lipoprotein, and macrophages in stable and unstable angina and acute myocardial infarction.

BACKGROUND: C reactive protein (CRP), an important serum marker of atherosclerotic vascular disease, has recently been reported to be active inside human atherosclerotic plaques. AIMS: To investigate the simultaneous presence of macrophages, CRP, membrane attack complex C5b-9 (MAC), and oxidised low density lipoprotein (oxLDL) in atherectomy specimens from patients with different coronary syndromes. METHODS: In total, 54 patients with stable angina (SA; n = 21), unstable angina (UA; n = 15), and myocardial infarction (MI; n = 18) underwent directional coronary atherectomy for coronary lesions. Cryostat sections of atherosclerotic plaques were immunohistochemically stained with monoclonal antibodies: anti-CD68 (macrophages), anti-5G4 (CRP), aE11 (MAC), and 12E7 (oxLDL). Immunopositive areas were evaluated in relation to fibrous and neointima tissues, atheroma, and media. Quantitative analysis was performed using image cytometry with systematic random sampling (percentage immunopositive/total tissue area). RESULTS: Macrophages, CRP, MAC, and oxLDL were simultaneously present in a higher proportion of fibrous tissue and atheroma of atherectomy specimens from patients with UA and MI compared with SA (p<0.05). Quantitative analysis showed significantly higher mean percentages of macrophages in plaques from patients with MI (44%) than UA (30%; p<0.01) and SA (20%; p<0.001). Significantly higher mean percentages of CRP were also seen in MI (25%) and UA (25%) compared with SA (12%; p<0.05). CONCLUSIONS: The presence of CRP, complement, and oxLDL in a high proportion of plaque tissue from patients with unstable coronary artery disease implies that these surrogate markers have important proinflammatory effects inside atherosclerotic plaques. This may increase vulnerability to plaque rupture and thrombosis, with subsequent clinical sequelae.