No evidence of Borrelia in cutaneous infiltrates of B-cell lymphomas with a highly sensitive, semi-nested real-time polymerase chain reaction targeting the 5S-23S intergenic spacer region.

BACKGROUND: The role of Borrelia in the development of skin lymphomas has been under discussion for decades. A similar association has been shown for Helicobacter pylori and gastric lymphomas (MALT type). Nevertheless, few molecular studies investigated Borrelia in skin lymphomas and the results are controversial. METHODS: We analysed 46 formalin-fixed, paraffin-embedded skin specimens of clincopathologically confirmed B-cell lymphomas (15 marginal zone lymphomas; 20 follicular lymphomas; three diffuse large B-cell lymphomas; eight secondary cutaneous infiltrates) taken from 36 patients from Northern Germany, an endemic area for Borrelia. Fifteen pseudolymphomatous lesions of cutaneous Borreliosis served as the control. Both groups were examined with a real-time (rt) PCR and a semi-nested PCR targeting the 5S-23S intergenic spacer region (IGS). A multiplex PCR was used to investigate B-cell clonality in all lymphomatous infiltrates (Biomed Primers). RESULTS: With both assays no Borrelia burgdorferi-specific DNA was identified in any of the B-cell lymphomas, while all 15 Borreliosis specimens gave a positive PCR result in the semi-nested PCR protocol, 12 were also positive in the rt PCR (P < 0.01). All B-cell lymphomas showed monoclonal IgH-Rearrangement. Analysis of cutaneous B-cell lymphomas from available studies including ours (n = 334) reveals an odds ratio <1. CONCLUSION: While some previous studies suggested an association between B. burgdorferi and the development of cutaneous B-cell lymphomas in endemic areas, we were unable to confirm this in our patients, despite a highly sensitive Borrelia PCR assay. Our results including meta-analysis of previous studies question the need for antibiotic therapy in patients with cutaneous B-cell lymphomas.

Reconsideration of the diagnostic criteria required for PCV2 reproductive disease.

Porcine circovirus type 2 (PCV2) causes a variety of clinical conditions including PCV2-associated reproductive disease (PCV2-RD) characterized by late term abortions and mummifications. The generally accepted diagnostic triad includes the presence of reproductive disorders, the histopathological finding of myocarditis, and detection of moderate to high viral loads within the heart tissue. A new threshold of 109 PCV2 genome equivalents (GE)/g heart tissue is suggested to fulfil the third criterion using the diagnostic settings of quantitative real time PCR and in situ hybridization of 30 fetal heart tissues. The need to identify histopathological lesions in fetal heart tissue appears to be invalid or overestimated in confirming a diagnosis of PCV2-RD, at least at the individual fetus level. The highest viral loads (1012 GE/g tissue) were detected in autolyzed and mummified piglets and were identified as PCV2d, although concurrent detection of PCV2d + a and PCV2d + b also occurred.

Mechanism of the historical and the ongoing Vulcanian eruptions of Ebeko volcano, Northern Kuriles.

Ebeko is one of the most active volcanoes of the Kurile island arc, producing frequent mild Vulcanian explosions with eruption clouds up to 5 km high. The volcano poses a serious threat to the Severo-Kurilsk town with a population of around 2500 inhabitants, located at a distance of only 7 km on a fan of the volcano's laharic deposits. Here, we report an overview of the activity of the volcano in the 20th-21st centuries and the results of our geological and petrological investigations of the ongoing Vulcanian eruption that started in 2016. We have found that eruptions of Ebeko span a range of mechanisms from purely magmatic to phreatic/hydrothermal. Three of its historical eruptions (the 1934-1935, 1987-1991, and the 2016-ongoing) involved fresh magma, while during the others (1967-1971, 2009-2011) fresh magma was not erupted. Juvenile material of the ongoing eruption represents highly crystalline and highly viscous (more than 108 pa s) low-silica (56-58 wt% SiO2) andesite. Historical data and our observations of the ongoing eruption allowed us to suggest a functional model of the volcano where Vulcanian explosions are caused by shallow intrusions of small diapir-like batches of strongly crystallized and highly viscous andesitic magma ascending into water-saturated, hydrothermally altered rocks composing the volcano summit. We suggest that the diapir's ascent is governed by their positive buoyancy. Some of the diapirs reach and breach the ground surface producing magmatic eruptions of Ebeko, while the others are stuck at the shallow subsurface level and feed intensive hydrothermal activity as well as phreatic eruptions of the volcano. Positive buoyancy of the diapirs is too weak to allow them to extrude high above the ground surface to form lava domes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00445-020-01426-z.

Single centre versus multi-centre pooled morbidity data in PCNL and the implications for informed consent.

Background: National and international outcome data following PCNL have been available for many years, but multi-centre data may not reflect the outcome from an individual surgeon or hospital.Methods: A combination of retrospective and prospective single centre data was collated from 2000-2016 and are compared to large single and multi-centre series.Results: Data were available on 801 unique cases performed between 2000 and 2016, mean age = 55.2 (SD = 14.8) (range = 17-93). The mean change in haemoglobin after PCNL was 1.65 g/dL ± 0.05, n = 630. Twenty-seven patients required a blood transfusion (3.37%). In 470 cases, data on pre-operative urine culture was available. One hundred and nineteen (25%) demonstrated evidence of bacteriuria pre-operatively. The most common isolated species were E. Coli and Proteus Mirabilis. Pre-operative urine infection was associated with a greater drop in haemoglobin following surgery, but this difference was not found to be statistically significant. Changes in serum creatinine and eGFR rise following surgery were calculated. The mean rise was found to be 15.21 µmol/L (SE = 2.08, n = 208). The mean drop in eGFR was estimated to be 7.35 ml/min/1.73 m2 (± 0.895, n = 205). Eight cases of 801 (1%) required admission to higher level care. There was one small bowel puncture and one pleural perforation recorded. Sub-selective embolization due to bleeding occurred in six cases (0 .75%) and there were no peri-operative deaths in this series. Published data comparing single centres with > 500 cases are presented.Conclusion: To facilitate transparent consent, single-centre rather than pooled outcome data should be utilized.

Detection of selected viral pathogens in dogs with canine infectious respiratory disease in Austria.

OBJECTIVES: To assess the prevalence of canine parainfluenza virus, canine adenovirus type 2, canine distemper virus, canine respiratory coronavirus and influenza virus A infections in: (1) privately-owned or, (2) kennelled dogs showing signs consistent with canine infectious respiratory disease and, (3) clinically healthy dogs in Vienna, Austria. MATERIALS AND METHODS: Prospectively, nasal and tonsillar swabs from 214 dogs affected with infectious respiratory disease, and 50 healthy control dogs were tested for nucleic acids specific to the various viral infections. Concurrent bronchoalveolar lavage fluid from 31 dogs with chronic respiratory disease was investigated for the same viral pathogens. Additionally, anti-canine respiratory coronavirus antibody concentrations were measured in paired blood samples from 30 acutely diseased dogs. RESULTS: Canine respiratory coronavirus (7.5%) and canine parainfluenza virus (6.5%) were the most commonly detected viruses in samples from the upper airways of dogs with respiratory infections. Serological results showed a significant seroconversion in response to coronavirus in 50% of the examined cases. None of the samples was positive for influenza virus A-specific nucleic acid. Canine coronavirus-specific nucleic acid was detected in 4.0% of healthy dogs. CLINICAL SIGNIFICANCE: Canine coronavirus should be considered as a clinically relevant cause of infectious respiratory disease in crowded dog populations. For sample collection, the nasal mucosa can be recommended as the favoured site. Analysis of paired serum samples aids verification of canine coronavirus infection in respiratory disease.

Ensuring selectivity using retention time modulation by solvent composition, applied to the analysis of phenol in e-liquids.

A gas chromatographic method for the determination of volatile contaminants (halogenated solvents, benzene, toluene, ethyl-benzene, xylenes, styrene) and phenol in e-liquids was developed and validated with a working range of 0.01 (limit of quantification) to 0.5 mg/l, and variation coefficients between 2 and 14%. Selectivity performing MS/MS-detection was sufficient for all analytes except for phenol: e-liquids contain high amounts of aroma compounds in excess of 105 compared to phenol. A number of these compounds potentially interfere at the retention time of phenol, showing all masses (including daughter ions and transitions) of phenol. To allow the detection of phenol in this matrix, a novel approach of adding a polar molecule to the injection solvent was used, modulating the polarity of the column, and thus the retention time of phenol. By adding 3 µl/ml and 10 µl/ml of 1,2-propanediol the retention time of phenol was shifted by 0.06 and 0.11 min respectively, while interfering peaks have not been shifted. This allowed a reliable confirmation of the presence of phenol. The introduced approach is an easy way to generate an additional chromatographic dimension for confirmation purposes, not requiring additional equipment.

[Skin biopsy of inflammatory skin diseases in childhood-when is it reasonable?] / Hautbiopsie bei entzündlichen Hauterkrankungen im Kindesalter ­ wann ist sie sinnvoll?

Compared with adulthood, inflammatory skin diseases are relatively rarely biopsied in children. Apart from the invasiveness of the procedure, the required local anesthesia, and the risks of infection and scarring, the psychological trauma of the operation has a higher impact in childhood. If a biopsy is performed, expectations towards the dermatopathology report are high. However, the evaluation of biopsies taken from children is challenging for the dermatopathologist: on the one hand, because the biopsies are often tiny or just superficial shaves and, on the other hand, because criteria for evaluation have mostly been developed from findings in adult biopsy specimens. In children, the immune system is still in the process of maturation and, therefore, infiltrates in the skin may look different from those seen in adults; however, knowledge about that is very limited to date. Moreover, numerous rare genodermatoses may manifest themselves first in childhood and need to be considered in the differential diagnosis while experience with them is often limited. Starting from the clinical presentation, this article presents histopathological features of possible differential diagnoses in order to demonstrate the value or necessity of a skin biopsy in a pediatric patient. In addition, communication with parents and child, methods of local anesthesia and biopsy techniques will be considered.

[Relevance of biopsies for the diagnostics of infectious skin diseases]. / Stellenwert der Biopsie in der Diagnostik von Infektionserkrankungen der Haut.

Pathogen-related skin infections are a common problem in the dermatological practice. Apart from culturing and serological detection methods, a skin biopsy is a possible diagnostic procedure, especially when the clinical picture is unspecific and other non-infectious skin diseases are considered as possible differential diagnoses. Some organisms can already be detected by routine staining methods (hematoxylin & eosin, e. g., yeasts, Leishmania), for others numerous histochemical and immunohistochemical stains are available, e. g. periodic acid-Schiff reaction (PAS) and Grocott for hyphae and spores, Ziehl-Neelson and Fite-Faraco for Mycobacteria or specific antibodies for Treponema pallidum or herpesviruses. In other instances, an infectious disease may not be diagnosed with certainty in a histological section but the pattern of inflammatory infiltrates is highly suggestive of an infectious cause. Based on such reaction patterns, the dermatopathologist can advise the clinician to perform cultures or serological investigations or additional molecular biological techniques can be applied to the biopsy specimen in order to identify the pathogens. This article presents skin infections with their histopathological features and highlights diseases that can be diagnosed with certainty in a biopsy and those in which a biopsy is helpful to exclude differential diagnoses or to perform molecular diagnostics on the specimen.

The role of dispersion type metalπ interaction in the enantiotropic phase transition of two polymorphs of tris-(thienyl)bismuthine.

Two polymorphs of tris(thienyl)bismuthine Bi(2-C4H3S)3 (1) were isolated upon crystallization from n-hexane at different temperatures. The high temperature form 1-HT crystallized at 269 K in the trigonal space group R3[combining macron], whereas the low temperature form 1-LT crystallized at 245 K in the triclinic space group P1[combining macron]. An enantiotropic phase transition was observed at 250 K showing a transition energy of 1.4 kJ mol-1. Both polymorphs reveal the formation of centrosymmetric dimers that are based on London dispersion type bismuthπ heteroarene interactions. These primary building units show additional dispersion type interactions between neighbouring dimers and as a result 2D networks are formed. DFT calculations on the model systems BiX3π thiophene (X = Cl, Me) verify the hypothesis of a soft and shallow binding potential of the London dispersion type bismuthπ heteroarene interaction, providing an explanation for the reversibility of the phase transition.

[Differential diagnosis of panniculitides]. / Differenzialdiagnostik der Pannikulitis.

Panniculitides are diseases of the subcutaneous tissue with heterogeneous etiology. They may develop consequent to infections, as a reaction to drugs, after thermal injury, as part of autoimmune diseases, in metabolic disorders or due to infectious organisms. The clinical presentation with subcutaneous nodules is often nonspecific. Moreover, the differentiation from vasculitides of medium-sized vessels can be clinically challenging. Microscopic examination of biopsy specimens is of high importance in the differential diagnosis of panniculitides. Histopathologically, panniculitides can be classified according to the predominantly infiltrated area in septal and lobular panniculitides and they can be separated from vasculitides of medium-sized vessels. Diagnostic difficulties arise from inadequate biopsy specimens and from lack of clinicopathological correlation. This article summarizes diagnostic criteria of frequent and clinically important panniculitides.

Species identification of dermatophytes in paraffin-embedded biopsies with a new polymerase chain reaction assay targeting the internal transcribed spacer 2 region and comparison with histopathological features.

BACKGROUND: Dermatophytosis is a very common skin infection with a broad clinical spectrum. Biopsies are often used to confirm the diagnosis, especially when the clinical presentation is unusual. Not uncommonly, organisms are hard to find even with periodic acid-Schiff stains. Polymerase chain reaction (PCR) for dermatophytes can be used in such cases. OBJECTIVES: To test a new PCR assay allowing species identification of dermatophytes on paraffin-embedded biopsies, and to reassess histopathological criteria for diagnosis of dermatophytosis. METHODS: In total, 121 biopsies of 92 patients with clinical suspicion of tinea were included. In 42 samples the clinical diagnosis had been confirmed histopathologically, and in 79 no fungal elements had been identified. PCRs targeting the internal transcribed spacer (ITS)2 region of dermatophytes were performed on the biopsies with subsequent sequencing. Sections were reassessed for the presence/absence of hyphae/spores, pattern and composition of infiltrate, and epidermal/follicular changes. Patient charts were reviewed for clinical data. RESULTS: The new ITS2 PCR assay detected 94% of the dermatophyte infections (compared with 79% identified by microscopy). Trichophyton rubrum was the dominant species (89%), and other species identified were Trichophyton verrucosum (2%), Microsporum canis (4%), Epidermophyton floccosum (2%) and Trichophyton interdigitale (4%). In particular, infections with T. interdigitale and manifestations with prominent spongiosis were not diagnosed histologically. Intracorneal neutrophils, which have been emphasized as a histopathological clue to dermatophytosis, were present in only 46% of PCR-positive samples. CONCLUSIONS: Molecular species identification of dermatophytes via ITS2 PCR can easily be implemented in a routine dermatopathology setting. It is fast and highly specific and improves the sensitivity of histopathological diagnosis of dermatophytosis.

Fast, sensitive and specific diagnosis of infections with Leishmania spp. in formalin-fixed, paraffin-embedded skin biopsies by cytochrome b polymerase chain reaction.

BACKGROUND: Northern spread of sandflies and Leishmania spp. has been observed in Europe. Diagnosis can be difficult owing to the various clinical manifestations. Species identification is important for patient management and therapy. Molecular diagnostics is increasingly used for pan-Leishmania detection but species identification remains challenging in formalin-fixed material. OBJECTIVES: To apply cytochrome b (cytb) polymerase chain reaction (PCR) and sequencing for identification of Leishmania species on formalin-fixed, paraffin-embedded (FFPE) skin biopsies; and to identify species-specific histological patterns. METHODS: Sixty-nine biopsies (48 patients) diagnosed with leishmaniasis based on the presence of amastigotes in the tissue (n = 41) or granulomatous infiltrates with positive pan-Leishmania real-time PCR (n = 28) were analysed with cytb PCR, sequencing and phylogenetic analysis. Histological sections were analysed; epidemiological data were collected. RESULTS: Cytb PCR identified Leishmania in all specimens: L. infantum (79%), L. major (8%), L. panamensis (4%), L. tropica (4%), L. killicki (2%) and L. aethiopica (2%). Of the detected species 95% were endemic to the country in which the infection was acquired. Amastigotes were found in 59%. Infiltrates were mainly tuberculoid granulomatous (65%), interstitial (15%) and sarcoidal (10%). Pseudolymphomatous features and pseudocarcinomatous hyperplasia were more common in L. major infections than in L. infantum (P < 0·01). CONCLUSIONS: Cytb PCR and sequencing is a fast, reliable and sensitive assay for identification of Leishmania spp. in FFPE biopsies. Leishmania infantum is the main cause of cutaneous leishmaniasis in Germany. Tuberculoid granulomas, other types of granulomas and pseudolymphomatous infiltrates may be encountered; the latter being indicative of infection with L. major.

[Cutaneous granulomatous reactions at botulinum neurotoxin A injection sites: First manifestation of systemic sarcoidosis]. / Granulomatöse Hautveränderungen an Botulinumtoxin-A-Injektionsstellen : Erstmanifestation einer systemischen Sarkoidose.

CASE REPORT: We report a case of granulomatous skin reaction after injection of an unknown botulinum neurotoxin (BoNT) A preparation. Four years after occurrence of skin lesions, systemic sarcoidosis became manifest. CONCLUSION: In addition to injection trauma, the BoNT A preparation may have acted as an immunogenic stimulus leading to cutaneous manifestation of sarcoidosis.

[Differential diagnostics of sebaceous tumors]. / Differenzialdiagnostik der Talgdrüsentumoren.

Sebaceous tumors are epithelial tumors with a differentiation towards sebaceous adnexal structures of the skin. They imitate the epithelial cells of mature sebaceous glands, sebaceous ducts, immature (embryonic) sebaceous structures or sebaceous glands that are not stimulated by hormones (mantle structures). This article explains the classification of sebaceous tumors on the basis of the normal histology of sebaceous glands. Clinical and histopathological criteria are given for the most important sebaceous tumors. The differential diagnosis of sebaceoma, sebaceous adenoma and various types of sebaceous carcinoma is emphasized. The importance of a specific diagnosis of adnexal tumors is demonstrated by tumor-associated syndromes with involvement of other organs (e.g., Muir-Torre syndrome and Birt-Hogg-Dubé syndrome). Furthermore, conceptional controversies, problems in differential diagnosis and the impact of immunohistochemical staining in the assessment of sebaceous tumors are considered.

[Fundamentals of cutaneous adnexal tumors]. / Basiswissen Hautadnextumoren.

Adnexal tumors of the skin are epithelial skin tumors with differentiation towards the adnexal epithelial structures of the skin, namely, hair follicle, sebaceous gland, apocrine gland, and eccrine gland. Adnexal tumors include hamartomas, benign and malignant neoplasms, and hyperplasias. The specific diagnosis of adnexal tumors is important because some lesions such as sebaceous neoplasms, cylindromas, or fibrofolliculomas are herald lesions of hereditary tumor syndromes (e.g., Muir-Torre syndrome, familial cylindromatosis, Birt-Hogg-Dubé syndrome). In this article, the classification of adnexal tumors of the skin is explained on the basis of embryology and histology and the main features of tumor-associated syndromes are summarized. Moreover, some conceptual controversies and problems in differential diagnosis of cutaneous adnexal tumors are discussed.

Genotyping of Borrelia from formalin-fixed paraffin-embedded skin biopsies of cutaneous borreliosis and tick bite reactions by assays targeting the intergenic spacer region, ospA and ospC genes.

BACKGROUND: Lyme borreliosis has a broad spectrum of clinical presentations involving the skin, joints and nervous system. The variable manifestations have been attributed to different Borrelia genospecies but genotyping required culture or fresh tissue. However, in dermatology practice, formalin-fixed paraffin-embedded biopsies are used for dermatopathological examination. Polymerase chain reaction (PCR) for Borrelia burgdorferi sensu latu has been established on such specimens, but studies attempting genotyping of subspecies or strains are lacking. OBJECTIVES: To adapt PCR assays for genotyping of Borrelia using paraffin-embedded biopsies, to identify Borrelia genospecies and to compare clinicopathological features of different genospecies. METHODS: Eighty-two paraffin-embedded biopsies from 68 patients, with erythema migrans, acrodermatitis chronica atrophicans, lymphocytoma cutis or tick bite reactions, were studied with assays targeting the intergenic spacer (IGS), ospA and ospC, followed by sequencing and phylogenetic analysis. Clinicopathological data were analysed comparing different Borrelia genospecies. RESULTS: Genotyping by IGS, ospA and ospC was successful in 85% of patients (91% B. afzelii, 7% B. garinii, 2% B. bavariensis). ospA serotyping identified type 2 (90%), type 3 (8%) and type 4 (2%). ospC-PCR was positive in 40% of the patients revealing 12 different groups, noninvasive forms being seen only in tick bite reactions and erythema migrans. No major clinicopathological differences could be identified between the genospecies, but neural inflammation and arthralgia were seen more often in lesions caused by invasive ospC strains. CONCLUSIONS: Genotyping of Borrelia can be easily implemented in a routine dermatopathology setting, especially as a fast method to confirm early cutaneous borreliosis. Genotyping could also enable earlier treatment of patients infected with invasive strains.

[Fundamentals of histology]. / Basiswissen Histologie.

Very commonly skin biopsies are crucial for the correct diagnosis of patients with skin diseases. The dermatopathology report is an important tool for communication between the clinical dermatologist and the dermatopathologist. This article explains the method of histopathological assessment of skin biopsies and summarizes the most important criteria for diagnosis of common skin diseases. The application of additional histochemical, immunohistochemical, and molecular biological methods is elucidated. The clinical dermatologist requires basic knowledge of dermatopathology to decide on the appropriate biopsy procedure. It also enables him to understand the dermatopathology report and to identify settings in which clinicopathological correlation is required.