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Background: Vesiculobullous disorders (VBDs) are extant with diverse clinical manifestations. Vesicles and bullae are fluid-filled cavities present within or beneath the epidermis. They are autoimmune blistering disorders in which autoantibodies are directed against target antigens present in the epidermis and dermo-epidermal junction. Objective: Evaluation of the various clinicodemographic profile of patients with a pattern of distribution (subtypes) of VBDs of the skin and assess the association between clinical aspects and histological changes in vesiculobullous lesions of the skin. Materials and Methods: The study material constituted 93 cases of VBDs out of 936 skin biopsies reported over two and a half years (January 2016 to June 2018) from the tertiary care center. A detailed history of the patients was taken, and a complete physical and dermatological examination with findings including clinical diagnosis was recorded. Histopathological examination (incisional/excisional/punch biopsy) was done in each case. The clinico-demographic evaluation was done and the results were correlated with histopathological findings. Results: Vesiculobullous lesions constituted 10.06% of all skin biopsies. The majority of cases were of pemphigus vulgaris (PV) 30 (32.25%) followed by 16 (17.2%) of bullous pemphigoid. In 83 cases (89.24%) histopathology findings were consistent with clinical diagnosis. Out of 34 cases that were diagnosed clinically as PV , the histopathological study proved 30 cases (88.23%) as PV. Conclusion: Vesiculobullous lesions of the skin are a heterogeneous group of disorders. It is essential to differentiate each pattern of subtype based on clinical examination and histopathological findings. Histopathological diagnosis with clinical correlation plays a major role in arriving at the diagnosis.

Mieke Hulens,1 Frans Bruyninckx,2 Dietmar Rudolf Thal,3,4 Ricky Rasschaert,5 Chris Bervoets,6– 8 Wim Dankaerts1 1Department of Rehabilitation Sciences, Musculoskeletal Rehabilitation Research Unit, Faculty of Kinesiology and Rehabilitation Sciences, KU Leuven, Leuven, Belgium; 2Department of Physical Medicine & Rehabilitation, EMG-Laboratory, University Hospitals of Leuven, Leuven, Belgium; 3Department of Imaging and Pathology, Laboratory of Neuropathology, and Leuven Brain Institute, KU Leuven, Leuven, Belgium; 4Department of Pathology, University Hospitals of Leuven, Leuven, Belgium; 5Department of Neurosurgery, AZ Rivierenland, Bornem, Belgium; 6Department of Neurosciences, Adult Psychiatry, KU Leuven, Leuven, Belgium; 7Department of Ophthalmology, University Hospitals of Leuven, Leuven, Belgium; 8Department Adult Psychiatry, University Psychiatric Center of KU Leuven, Leuven, BelgiumCorrespondence: Mieke HulensDepartment of Rehabilitation Sciences, Musculoskeletal Rehabilitation Research Unit, Faculty of Kinesiology and Rehabilitation Sciences, University of Leuven, Tervuursevest 101, Leuven, Belgium, Tel +32 478 338003, Fax +32 16 32 91 97, Email miekehulens@gmail.comPurpose: Tarlov cysts (TCs) are dilated nerve root sheaths originating from increased cerebrospinal pressure. Patients with TCs often complain of neuropathic pain and paresthesia. The aim of this study was to retrospectively review intraepidermal nerve fiber density (IENFD) and electrodiagnostic (EDX) data from TC patients.Patients and Methods: Lower leg skin biopsy results and EDX data from the L2–S4 myotomes of patients with lumbar or sacral TCs ≥ 8 mm were retrieved from a database of a physical medicine clinic. Patients with compressive pathology, diabetes mellitus and chemotherapy were excluded.Results: IENFD data from 17 patients and EDX data from 24 patients with TCs ≥ 8 mm were available. The mean age was 47 ± 10y, and 83% were women. In 82% of patients, the IENFD was below the 5th percentile by age and sex. EDX showed increased Hoffmann reflex latencies in 25%, increased anal reflex latencies in 95%, and a patchy distribution of neurogenic motor unit potentials in 100%. More than 50% of needle EMG abnormalities appeared in myotomes unrelated to the location of the TCs.Conclusion: Small- and/or large-fiber neuropathy was documented in a significant proportion of patients with TCs. The novel findings may add to the understanding of the mechanisms involved in symptomatic TCs. We propose that pathologically elevated cerebrospinal fluid pressure not only dilates some of the nerve root sheaths to form TCs but also potentially damages axons in nondilated nerve root sheaths and neurons in the dorsal root ganglia.Keywords: perineural cysts, skin biopsies, radicular pain, anal reflex, nerve conduction studies, fibromyalgia

Background: Obtaining high quality RNA from skin biopsies is complex due the physical composition and high content of nucleases of this tissue. This becomes particularly challenging when using compromised skin samples with necrotic, inflammed or damaged areas, such as those from patients suffering skin conditions, which affect more than 900 million people annually. We evaluated the impact of the biopsy size and tissue preservation method on the quality and quantity of RNA extracts. Methods: Skin lesion biopsies were obtained from patients with cutaneous leishmaniasis (CL). Biopsy specimens of 2 mm (n = 10) and 3 mm (n = 59) were preserved in Allprotect® reagent, and 4 mm biopsies in OCT (n = 54). Quality parameters were evaluated using Nanodrop and Bioanalyzer. The informativeness of the extracted samples for downstream analyses was evaluated using RT-qPCR and RNA-Seq. Results: The success rate, based on quality parameters of RNA extraction from tissue biopsies stored in OCT and 2 mm biopsies stored in Allprotect®, was 56% (30/54) and 30% (3/10), respectively. For 3 mm skin biopsies stored in Allprotect® was 93% (55/59). RNA preparations from 3 mm-Allprotect® biopsies had an average RIN of 7.2 ± 0.7, and their integrity was not impacted by sample storage time (up to 200 days at -20°C). RNA products were appropriate for qRT-PCR and RNA-seq. Based on these results, we propose a standardized method for RNA extraction from disrupted skin samples. This protocol was validated with lesion biopsies from CL patients (n = 30), having a success rate of 100%. Conclusions: Our results indicate that a biopsy size of 3 mm in diameter and preservation in Allprotect® for up to 200 days at -20°C, are best to obtain high quality RNA preparations from ulcerated skin lesion biopsy samples.

Leishmania infantum is a protozoan causing human zoonotic visceral leishmaniasis (ZVL) and visceral–cutaneous canine leishmaniosis (CanL) in the Mediterranean Basin. L. infantum is able to infect a large number of wild and domestic species, including cats, dogs, and horses. Since the 1990s, clinical cases of equine leishmaniasis (EL), typically characterized by cutaneous forms, have been increasingly diagnosed worldwide. The aim of the present study was to evaluate the presence of clinical forms of EL in CanL-endemic areas in Italy, where exposure of equine populations was ascertained from recent serological surveys. For this purpose, formalin-fixed and paraffin-embedded skin biopsies of 47 horses presenting chronic dermatitis compatible with EL were retrospectively selected for the study and subjected to conventional and q-PCR. A singular positivity for L. infantum was found; BLAST analysis of sequence amplicons revealed a 99–100% homology with L. infantum sequences. The histological examination revealed a nodular lymphoplasmacytic and histiocytic infiltrate; immunohistochemistry showed rare macrophages containing numerous positive amastigotes. The present retrospective study reports, for the first time, a case of a cutaneous lesion by L. infantum occurring in an Italian horse. Pathological and healthy skin samples should be investigated on a larger scale to provide information on the potential clinical impact of EL in the practice, and to define the role of horses in epidemiological ZVL and CanL scenarios.

ABSTRACT Human skin microbiota has been described as a “microbial fingerprint” due to observed differences between individuals. Current understanding of the cutaneous microbiota is based on sampling the outermost layers of the epidermis, while the microbiota in the remaining skin layers has not yet been fully characterized. Environmental conditions can vary drastically between the cutaneous compartments and give rise to unique communities. We demonstrate that the dermal microbiota is surprisingly similar among individuals and contains a specific subset of the epidermal microbiota. Variability in bacterial community composition decreased significantly from the epidermal to the dermal compartment but was similar among anatomic locations (hip and knee). The composition of the epidermal microbiota was more strongly affected by environmental factors than that of the dermal community. These results indicate a well-conserved dermal community that is functionally distinct from the epidermal community, challenging the current dogma. Future studies in cutaneous disorders and chronic infections may benefit by focusing on the dermal microbiota as a persistent microbial community. IMPORTANCE Human skin microbiota is thought to be unique according to the individual's lifestyle and genetic predisposition. This is true for the epidermal microbiota, while our findings demonstrate that the dermal microbiota is universal between healthy individuals. The preserved dermal microbial community is compositionally unique and functionally distinct to the specific environment in the depth of human skin. It is expected to have direct contact with the immune response of the human host, and research in the communication between host and microbiota should be targeted to this cutaneous compartment. This novel insight into specific microbial adaptation can be used advantageously in the research of chronic disorders and infections of the skin. It can enlighten the alteration between health and disease to the benefit of patients suffering from long-lasting socioeconomic illnesses.

Lumpy skin disease (LSD) diagnosis is primarily based on clinical surveillance complemented by PCR of lesion crusts or nodule biopsies. Since LSD can be subclinical, the sensitivity of clinical surveillance could be lower than expected. Furthermore, real-time PCR for the detection of LSD viral DNA in blood samples from subclinical animals is only intermittently positive. Therefore, this study aimed to investigate an acceptable, easily applicable and more sensitive testing method for the detection of clinical and subclinical LSD. An animal experiment was conducted to investigate ear notches and biopsies from unaffected skin taken from the neck and dorsal back as alternatives to blood samples. It was concluded that for early LSD confirmation, normal skin biopsies and ear notches are less fit for purpose, as LSDV DNA is only detectable in these samples several days after it is detectable in blood samples. On the other hand, blood samples are less advisable for the detection of subclinical animals, while ear notches and biopsies were positive for LSD viral DNA in all subclinically infected animals by 16 days post infection. In conclusion, ear notches could be used for surveillance to detect subclinical animals after removing the clinical animals from a herd, to regain trade by substantiating the freedom of disease or to support research on LSDV transmission from subclinical animals.

Introduction: Leprosy also known as Hansen’s disease is one of the oldest disease known to mankind. It is a chronic infectious disease caused by Mycobacterium leprae. It still remains as a major public health problem facing India. Nearly, 60% of all world leprosy cases are reported from India. The spectrum of presentation of leprosy is very wide. Histopathology is an important tool in making a definitive diagnosis. The objective of the study is to describe the histopathological profile of leprosy in the state of Bihar. Aim: To study the histopathological spectrum of leprosy at a tertiary care centre of Bihar. Materials and Methods: A retrospective hospital-based study of clinically diagnosed leprosy cases was conducted over a period of 28 months (January 2015 to April 2017). Lesional skin biopsies obtained were fixed, processed and stained with Haematoxylin and Eosin (H&E) followed by Ziehl-Neelsen (ZN) staining. The lesions were classified on microscopy as per Ridley-Jopling classification. Results: A total of 200 cases were studied. Highest incidence was in the 11 to 30 years age group for both males and females. Males were more affected (M:F=2.4:1). Most common clinical feature was loss of sensation. The commonest reported histopathological type was borderline lepromatous (43%) followed by borderline (17%). Overall ZN staining was positive in 56 (28%) cases. Conclusion: The spectrum of presentation of leprosy is very wide and there is clinical overlap between different types of leprosy. Histopathology still remains the gold standard for early diagnosis and classification of the disease. Accurate diagnosis forms the backbone for appropriate treatment and preventing deformities and drug resistance.

Screening for systemic amyloidosis is typically carried out with abdominal fat aspirates with varying reported sensitivities. Fat aspirates are preferred for use in primary screening instead of organ biopsies as they are less invasive and thereby minimize the potential risk of complications. At Odense Amyloidosis Center, we performed a prospective study on whether the combined use of fat aspirate and tru-cut skin biopsy could increase the diagnostic sensitivity. Both fat aspirates and skin biopsies were screened with Congo Red staining, and positive biopsies were subsequently subtyped using immunoelectron microscopy and mass spectrometry. Seventy-six patients were included. In total, 24 patients had systemic amyloidosis (11 AL, 12 wtATTR, 1 AA), and 6 patients had localized amyloidosis. Combined fat aspirate and skin biopsy were Congo Red-positive in 15 patients (overall sensitivity (OS) 62.5%). Fat aspirates were positive in 14 patients (OS 58.3%), and the skin biopsy was positive in 5 patients (OS 20.8%). In only one patient did the skin biopsy add extra diagnostic information. The sensitivity differed between AL and ATTR amyloidosis—81.8% and 41.7%, respectively. Using skin biopsy as the only screening method is not recommended.

Haematobia irritans exigua, commonly known as buffalo fly (BF), causes economic losses of about AUD $100 million per annum to the Australian cattle industry in terms of decreased production and costs of control. Lesions associated with BF infestation range from raised, dry, alopecic, hyperkeratotic or scab encrusted to severe hemorrhagic areas of ulceration which represent a major animal welfare concern. BF transmits a filarial nematode, Stephanofilaria sp., which has been speculatively associated with BF lesion development. The existing literature indicates that the sensitivity of currently used diagnostic techniques to detect Stephanofilaria in skin lesions is low and that there is currently no sequence for Stephanofilaria available on GenBank. Our objective is to develop a PCR method to detect Stephanofilaria in BF lesions. Skin biopsies were collected from 10 freshly slaughtered cattle hides having obvious BF eye lesions. Samples were collected from the center and the edge of the BF lesion as well as from adjacent normal tissue. Each skin punch was cut into 5-6 slices and immersed in normal saline before incubation overnight at 22°C. Eight nematodes were recovered from the saline by microscopic examination and preserved in ethanol. Nematode DNA will be extracted using conventional extraction methods. Specific primers will be used to amplify the ITS regions of rDNA and coxI region of the mtDNA and the amplified DNA will be sequenced. Primers will be designed from these regions to detect the presence of Stephanofilaria and used in PCR studies to clarify the etiology and epidemiology of BF lesions.

Background: Skin diseases are common in childhood and they are common reasons for pediatric visits to the hospital. In spite of this high occurrence, there are very few prospective studies addressing this issue. Aims: The present study was directed at determining the spectrum of dermato-pathological lesions encountered in a large general tertiary care hospital, over a two-year period. Materials and Methods: 107 cases formed the total sample studied, in a part prospective and part retrospective study. A detailed clinical history was recorded on a proforma prepared for the purpose and gross photographs were taken wherever possible. Results: Skin biopsies accounted for 7.29% of the total surgical pathology load, 55.44% of the total pediatric biopsies and 10.82% of the total number of skin biopsies. The age and sex distribution pattern revealed that the maximum number of biopsies (62.61%) were of older children, with a male preponderance (57.94%). The anatomic distribution pattern indicated predominant involvement of the limbs (59.82%). The maximum numbers of cases were of infectious nature (24.29%); the most frequently encountered being borderline tuberculoid Hansen′s disease (8.4%). A positive correlation with the clinical diagnosis was obtained in 56.07% cases. Conclusions: Histopathology contributed to the diagnosis in a significant number of (82.23%) cases, indicating its importance and utility.