Study protocol P-MAPS: microbiome as predictor of severity in acute pancreatitis-a prospective multicentre translational study.

BACKGROUND: Acute pancreatitis (AP) is an inflammatory disorder that causes a considerable economic health burden. While the overall mortality is low, around 20% of patients have a complicated course of disease resulting in increased morbidity and mortality. There is an emerging body of evidence that the microbiome exerts a crucial impact on the pathophysiology and course of AP. For several decades multiple clinical and laboratory parameters have been evaluated, and complex scoring systems were developed to predict the clinical course of AP upon admission. However, the majority of scoring systems are determined after several days and achieve a sensitivity around 70% for early prediction of severe AP. Thus, continued efforts are required to investigate reliable biomarkers for the early prediction of severity in order to guide early clinical management of AP patients. METHODS: We designed a multi-center, prospective clinical-translational study to test whether the orointestinal microbiome may serve as novel early predictor of the course, severity and outcome of patients with AP. We will recruit 400 AP patients and obtain buccal and rectal swabs within 72 h of admission to the hospital. Following DNA extraction, microbiome analysis will be performed using 3rd generation sequencing Oxford Nanopore Technologies (ONT) for 16S rRNA and metagenomic sequencing. Alpha- and beta-diversity will be determined and correlated to the revised Atlanta classification and additional clinical outcome parameters such as the length of hospital stay, number and type of complications, number of interventions and 30-day mortality. DISCUSSION: If AP patients show a distinct orointestinal microbiome dependent on the severity and course of the disease, microbiome sequencing could rapidly be implemented in the early clinical management of AP patients in the future. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04777812.

α-Tanycytes of the adult hypothalamic third ventricle include distinct populations of FGF-responsive neural progenitors.

Emerging evidence suggests that new cells, including neurons, can be generated within the adult hypothalamus, suggesting the existence of a local neural stem/progenitor cell niche. Here, we identify α-tanycytes as key components of a hypothalamic niche in the adult mouse. Long-term lineage tracing in vivo using a GLAST::CreER(T2) conditional driver indicates that α-tanycytes are self-renewing cells that constitutively give rise to new tanycytes, astrocytes and sparse numbers of neurons. In vitro studies demonstrate that α-tanycytes, but not ß-tanycytes or parenchymal cells, are neurospherogenic. Distinct subpopulations of α-tanycytes exist, amongst which only GFAP-positive dorsal α2-tanycytes possess stem-like neurospherogenic activity. Fgf-10 and Fgf-18 are expressed specifically within ventral tanycyte subpopulations; α-tanycytes require fibroblast growth factor signalling to maintain their proliferation ex vivo and elevated fibroblast growth factor levels lead to enhanced proliferation of α-tanycytes in vivo. Our results suggest that α-tanycytes form the critical component of a hypothalamic stem cell niche, and that local fibroblast growth factor signalling governs their proliferation.

Effect of ultraviolet (UV) A, UVB or ionizing radiation on the cell cycle of human melanoma cells.

BACKGROUND: One important component of the cellular response to irradiation is the activation of cell cycle checkpoints. It is known that both ultraviolet (UV) radiation and ionizing radiation (IR) can activate checkpoints at transitions from G(1) to S phase, from G(2) phase to mitosis and during DNA replication. OBJECTIVES: To evaluate the effects of irradiation with different wavelengths on cell cycle alterations. METHODS: p53-deficient IPC-298 melanoma cells were irradiated with 10 J cm(-2) UVA, 40 mJ cm(-2) UVB, or with 7.5 Gy IR. Cell cycle effects were then determined by DNA/5-bromodeoxyuridine dual-parameter flow cytometry. RESULTS: IPC-298 cells irradiated in G(1) with UVA were not arrested at the G(1)/S transition, but at the G(2)/M transition. Despite p53 deficiency, the cells showed a G(1) arrest after UVB exposure. Furthermore, IR did not affect G(1) or S phase, but induced G(2) phase arrest. Hence, the effects of UVA, but not of UVB, on the cell cycle in p53-deficient melanoma cells are comparable with those of IR. CONCLUSIONS: UVA and IR induce radical-mediated strand breaks and DNA lesions, and UVB essentially induces thymine dimers that lead to excision repair-related strand breaks. Different cell cycle effects may be a consequence of different types of DNA damage. The results showed that UVB-irradiated p53-deficient cells are arrested in G(1). Irradiation with the solar radiation component UVB can therefore result in a beneficial retardation of tumour promotion in human skin carrying p53-mutated cell clones.

Malignant melanoma of the anal region.

Malignant melanoma (MM) of the anal region is an uncommon disease. In many cases, the disease is undetected or mistaken for a benign polyp or haemorrhoids until it reaches an advanced state. Owing to delayed diagnosis and early metastases, the prognosis is often poor. In contrast to melanomas of the skin, a history of sun exposure does not seem to have an impact in development of MM in this region. Anorectal melanomas (AM) are most common in the rectum, followed by the anal canal and anal verge. Ras mutations, especially in codon 61 of the N-ras oncogene, are common in CM and rare in melanomas of the vulva and anorectum. The diagnosis of an AM is usually made using a biopsy. Histopathological examinations show spindle-shaped and pleomorphic cells. Adjuvant immunohistological markers are the calcium-binding protein S-100, the melanoma antigen HMB-45, the melanoma-expressed protein Melan A, and microphthalmia-associated transcription factor (MiTF). To date, there are few published guidelines for the correct management of AM, and surgery remains the mainstay of treatment. We report on a 39-year old man who presented with a 5-week history of recurrent prolapse of an anal tumour. The tumour was histologically confirmed to be malignant melanoma.

Perniosis-like tinea corporis caused by Trichophyton verrucosum in cold-exposed individuals.

Trichophyton verrucosum is a zoophilic infectious agent causing 98% of the dermatophytic infections of cattle. Transmission to humans has, until recently, been rare. One reason for an increase of infection in humans and animals seems to be the decrease in immunisation of cattle. We report on three cases of pertinent human infections with disseminated, sharply defined, bluish red, partly oedematous nodules and plaques in particular not only on the thighs, but also on the trunk and arms. Two of our patients work with farm animals. The third one works as an assistant in a butcher shop, but lives on a cow farm. All three patients are often exposed to the cold. In all three cases T. verrucosum was detected by culture. Tinea corporis was histologically confirmed in two patients. Based on the microbiological results, we began a combined systemic and local antimycotic therapy with fluconazole 50 mg day(-1) in two patients, itraconazole 100 mg day(-1) in one patient p.o. combined with topical ciclopiroxolamine. All patients were cured. Dermatophytosis caused by T. verrucosum can, under certain circumstances, such as frequent exposure to cold or a long-term corticosteroid therapy, mimic the characteristic clinical picture of perniosis, as we demonstrate here.

Type 1 diabetes mellitus in childhood: a matched case control study in Lancashire and Cumbria, UK.

AIMS: The aim of the study was to identify environmental risk factors for insulin-dependent diabetes mellitus (Type 1 DM) in childhood. METHODS: A matched case-control study of Type 1 DM conducted in Lancashire and Cumbria, UK, using a structured interview. Cases (n=196, participation rate 83%) were children under 16 years of age diagnosed prior to October 1998 and attending diabetic clinics. Controls (n=381) were healthy children from the community matched by gender and by age (within a few days of birth). The data were analysed by logistic regression using the technique of Breslow and Day for matched case control studies. RESULTS: The multivariate regression model showed that the following factors were significantly associated with the risk of developing Type 1 DM (odds ratio, 95% confidence intervals): sharing a room with a sibling (0.458, 0.290-0.721), social contact with other children when aged 6-11 months (0.439, 0.256-0.752), consumption of sugary food (0.080, 0.024-0.261), parental insulin dependent diabetes mellitus (10.651, 3.086-36.761), maternal thyroid disease (4.861, 1.681-14.058), consuming more than one pint of milk per day prior to school entry (0.498, 0.310-0.802), maternal smoking during pregnancy (0.373, 0.218-0.636), a father with no academic qualifications (0.504, 0.278-0.913), maternal age at time of birth (0.900, 0.854-0.948), maternal infections in pregnancy (2.453, 1.011-5.948), other maternal illnesses or conditions in pregnancy (2.007, 1.139-3.535), belonging to an Asian family (0.104, 0.028-0.394), and regular contact with pets and other animals (0.552, 0.309-0.987). CONCLUSION: Many of the results are consistent with the hygiene hypothesis which links improved living standards with decreased exposure to microorganisms and increased risk of immune mediated disease in childhood. These findings challenge the idea that improved hygiene acts exclusively through a Th2 mechanism leading to atopic disease as Type 1 DM is mediated by a Th1 reaction. The association with maternal smoking could be due to recall bias but a causal link cannot be excluded with confidence.

Tobacco smoke is phototoxic.

BACKGROUND: Both cigarette smoke and ultraviolet (UV) radiation are known to cause changes of the skin which can be regarded as premature ageing. OBJECTIVES: To assess the theory that the effects of these two exposures could be linked by a phototoxic action of cigarette smoke. METHODS: A photohaemolysis test was used, in which human erythrocytes were incubated with cigarette smoke condensate, followed by UV irradiation and measurement of exposure-dependent haemolysis. RESULTS: Cigarette smoke condensate was clearly phototoxic. Photohaemolysis depended on the concentration of the condensate and UV dose and was more pronounced after exposure to UVA-rich than UVB-rich radiation. CONCLUSIONS: Phototoxicity may be a mechanism by which cigarette smoking causes premature skin ageing. An enhancing effect on photocarcinogenesis has also to be considered.

Congenital adrenal hyperplasia and acne in male patients.

Seborrhoea is one pathogenic factor for acne. Androgens induce sebum production, and excess androgen may provoke or aggravate acne. In women an androgen disorder is frequently suspected when acne is accompanied by hirsutism or menstrual irregularities. In men acne may be the only symptom of androgen excess. We report three male acne patients in whom hormonal screening revealed irregularities of androgen metabolism suggestive of late-onset congenital adrenal hyperplasia and who benefitted from low-dose glucocorticoids. Disorders of androgen metabolism may influence acne not only in women, but also in men, and these patients may benefit from low-dose glucocorticoid therapy.

A polymorphism in the dopamine receptor DRD5 is associated with blepharospasm.

Abnormalities in dopamine neurotransmission are thought to underlie the generation of dystonic movements. The authors performed a case-control allelic association study in patients with the focal dystonia blepharospasm, using polymorphisms in the dopamine receptor and transporter genes. Allele 2 of a DRD5 dinucleotide repeat was significantly associated with blepharospasm. This may indicate a pathogenic role for this receptor.

Vertebrate development: Et in Arkadia.

The similarities in organiser formation in Xenopus and mouse embryos have remained elusive. Recent evidence suggests a common mechanism, in which an intracellular protein, Arkadia, is required for formation of the mouse organiser and potentiates the effects of the signalling protein Nodal.

Cervical dystonia is associated with a polymorphism in the dopamine (D5) receptor gene.

The objective was to assess whether polymorphisms in the dopamine receptor and transporter genes are associated with development of primary cervical dystonia. A case-control allelic association study is described of 100 patients with cervical dystonia and 100 controls using polymorphisms within D1-5 receptor and dopamine transporter genes. No significant association was found between patient and control allele frequencies for polymorphisms in genes for the D1 to 4 receptors and dopamine transporter. Significant associations, however, were found for alleles 2 and 6 of the D5 receptor microsatellite. Carriage of allele 2 was associated with cervical dystonia, whereas allele 6 was overrepresented in the control group, implying a possible protective effect. The association with allele 6 remained significant after Bonferroni correction. In conclusion, the finding of a significant association with an allele in the D5 receptor gene in patients with cervical dystonia may indicate a pathogenic role of this gene (or neighbouring genes). Further studies are required to confirm this finding and to assess whether these alleles are part of distinct haplotypes associated with other polymorphisms imparting a functional effect on the D5 receptor.

The role of Sonic hedgehog in neural tube patterning.

In the developing neural tube of vertebrate embryos, many types of neuronal and nonneuronal cells differentiate in response to the secreted signalling molecule, Shh. Shh shows a spatially restricted pattern of expression in cells located at the ventral midline, yet governs the differentiation of diverse cell types throughout the ventral half of the neural tube. Here, we describe how the distinct fate assumed by cells in response to Shh is dependent upon their position with respect to both the dorso-ventral and anterior-posterior axes of the neural tube and describe the ways in which a single factor, Shh, is able to pattern the developing nervous system. We first discuss the evidence that Shh does impose ventral identity on cells in the neural tube, then focus on the role of a graded Shh signal in patterning the neural tube and finally discuss the interaction of Shh with other factors that affect its signalling outcome.

Development of chick axial mesoderm: specification of prechordal mesoderm by anterior endoderm-derived TGFbeta family signalling.

Two populations of axial mesoderm cells can be recognised in the chick embryo, posterior notochord and anterior prechordal mesoderm. We have examined the cellular and molecular events that govern the specification of prechordal mesoderm. We report that notochord and prechordal mesoderm cells are intermingled and share expression of many markers as they initially extend out of Hensen's node. In vitro culture studies, together with in vivo grafting experiments, reveal that early extending axial mesoderm cells are labile and that their character may be defined subsequently through signals that derive from anterior endodermal tissues. Anterior endoderm elicits aspects of prechordal mesoderm identity in extending axial mesoderm by repressing notochord characteristics, briefly maintaining gsc expression and inducing BMP7 expression. Together these experiments suggest that, in vivo, signalling by anterior endoderm may determine the extent of prechordal mesoderm. The transforming growth factor (beta) (TGFbeta) superfamily members BMP2, BMP4, BMP7 and activin, all of which are transiently expressed in anterior endoderm mimic distinct aspects of its patterning actions. Together our results suggest that anterior endoderm-derived TGFbetas may specify prechordal mesoderm character in chick axial mesoderm.

Systemic vitamin C and vitamin E do not prevent photoprovocation test reactions in polymorphous light eruption.

The possible influence of oxidative stress is discussed in the pathogenesis of polymorphous light eruption (PLE). A double-blind, placebo-controlled study of prophylactic treatment with systemic administration of vitamin C (3 g/d) and E (1500 IU/d) for 8 days was undertaken in 9 patients with PLE (verum, n=4; placebo, n=5). Evaluation of the maximal effects after photoprovocation before and after intake of the antioxidants revealed a reduction of most skin reactions (overall skin reaction, papules/vesicles) in both groups with marked differences in the placebo group. The antioxidants in the doses given and over the time period used did not influence the development of PLE, but might interfere with immunosuppressive effects of repeated photoprovocation tests.