Polyethylene glycol-based isolation of urinary extracellular vesicles, an easily adoptable protocol.

Urine is a highly advantageous biological specimen for biomarker research and is a non-invasive source. Most of the urinary biomarkers are non-specific, volatile and need extensive validation before clinical adoption. Extracellular vesicles are secreted by almost all cells and are involved in homoeostasis, intercellular communication, and cellular processes in healthy and pathophysiological states. Urinary extracellular vesicles (UEVs) are released from the urogenital system and mirror the molecular processes of physiological and pathological states of their source cells. Therefore, UEVs serve as a valuable source of biomarkers for the non-invasive diagnosis of various pathologies. They hold a promising source of multiplex biomarkers suitable for prognosis, diagnosis, and therapy monitoring. UEVs are easily accessible, non-invasive, and suited for longitudinal sampling. Although various techniques are available for isolating UEVs, there is yet to be a consensus on a standard and ideal protocol. We have optimized an efficient, reliable, and easily adoptable polyethylene glycol (PEG) based UEV isolation technique following MISEV guidelines. The method is suitable for various downstream applications of UEVs. This could be a cost-effective, consistent, and accessible procedure for many clinical labs and is most suited for longitudinal analysis. Adopting the protocol will pave the way for establishing UEVs as the ideal biomarker source. •Urine can be collected non-invasively and repeatedly, hence a very useful specimen for biomarker discovery. Urinary EVs (UEVs), derived from urine, offer a stable diagnostic tool, but standardised isolation and analysis approaches are warranted.•To have enough UEVs for any study, large volumes of urine sample are necessary, which limits different isolation methods by cost, yield, and time.•The protocol developed could help researchers by offering a cost-effective and dependable UEV isolation method and may lay the foundation for UEVs adoption in clinical space.

The fatal fetal tumor: a geneticist's perspective.

Epignathus is an extremely rare oral teratoma which leads to high mortality in the early neonatal period. Various theories have been put forward for the genesis of such a tumor, though none is completely convincing. A genetic basis is not well established for the tumor. Microdeletions/duplications, as well as single gene disorders, have been known to cause epignathus, all with additional malformations. Evidence of single gene involvement in an isolated epignathus is lacking. We present a case of a 19-week-fetus with oro-pharyngeal teratoma detected on the level II ultrasound. The couple was counseled regarding the grave prognosis of the fetal condition following which they opted for termination of pregnancy and fetal autopsy. The autopsy revealed fetus-like body attached to the tumor. Genetic testing including a whole genome microarray did not reveal any significant variant. An explanation for the fetus-like body maybe a common origin of the teratoma and the additional fetus-like bodies due to an erroneous process of early embryonic development. Another possibility is of an acardiacus acranius twin masquerading as a fetus-like body. Thus, we conclude that in the absence of an associated malformation, an epignathus is unlikely to have a genetic etiology. This study highlights the importance of performing a fetal autopsy as a part of deep phenotyping to ascertain the etiology, as it identified additional fetal-like body which was not detected on the antenatal ultrasound.

Histopathology of Pemphigus Vulgaris Revisited.

ABSTRACT: Pemphigus vulgaris (PV) is a severe, potentially life-threatening autoimmune blistering disease, which is common in India. Although there is abundant literature on clinical and immunologic features, comprehensive studies on its histopathology are lacking. The aim of this study was to describe the histopathologic and immunofluorescence features as well as discuss various diagnostic pitfalls of PV. Histopathologic and immunofluorescence (DIF/IIF) findings were reviewed for 169 biopsies from 2007 to 2017 (11 years). The 169 samples included 152 skin, 16 oral mucosal, and 1 corneal biopsy. Maximum prevalence was noted in the fifth decade (57%) with a slight male preponderance. Vesicles were seen in 149 cases (88%), the level of which was suprabasal in 91(61%) and both suprabasal and intraepidermal in 50 cases (33.5%). Acantholytic cells were present in 142 cases (95%). Acantholytic keratinocytes showed rounded and polygonal acantholysis. 86 (51%) cases showed evidence of regeneration. Adnexal involvement was seen in 92 cases, commonest in the hair follicles. Unusual histologic findings included: intraepidermal bulla, absence of dermal inflammation, free floating hair shafts, multinucleated epithelial cells, eosinophil predominance; all of which are discussed. DIF was performed in 166 cases, of which 163 were positive (98%), and IgG was the commonest immunoreactant (96%). IIF was performed in 11 cases, of which 9 cases were positive for Dsg3. Although the diagnosis of PV rests on combined clinical, histologic, and IF features, histopathology as the sole means is also a powerful tool. It is important to be aware of the diagnostic pitfalls to optimize its utility.

An unusual case of hyperparathyroidism, presenting as multiple skeletal lesions.

We report the case of a 43-year-old male patient, who presented with severe osteoporosis and pain at multiple sites and a chest X-ray suggestive of few cystic lesions in bilateral ribs. Subsequently, the patient was referred for bone scan, in view of the suspicion of polyostotic fibrous dysplasia or neoplastic lesions. Bone scan showed homogenously increased tracer uptake in the axial and appendicular skeletal system, suggestive of metabolic superscan. Computed tomography neck localized right inferior parathyroid adenoma and expansile lytic skeletal lesions suggestive of brown tumors were also seen. This case highlights the importance of bone scan, helping in the differential diagnosis of multiple skeletal lesions.

Isolated Extranodal Rosai-Dorfman Disease on 18F-FDG PET-CT Scan.

Rosai-Dorfman disease (RDD) is an uncommon proliferative histiocytic disorder. Patients usually present with painless massive cervical lymphadenopathy with fever and leukocytosis. Isolated extranodal disease is rare and more severe fibrosis, fewer histiocytosis in lesions make diagnosis more difficult as compared to nodal disease. Here, we report a case of isolated extranodal RDD on fluorodeoxyglucose (FDG) positron emission tomography-computed tomography (PET-CT) scan. FDG-avidity of RDD lesions is attributable to the intense glucose dependence of the proliferating histiocytes. PET-CT scan not only demonstrates the complete staging of the disease but also provide functional information about the disease activity to guide biopsy.

Cutaneous Metastasis from Visceral Organs: 18F-Fluorodeoxyglucose Positron Emission Tomography-Computed Tomography Scan Aiding in Localizing Primary Site.

Skin metastases are rare from systemic malignancy, and incidence reported is between 0.7% and 9% in various malignancies and usually occur in advanced stage. Here, we report three cases, one each of carcinoma esophagus, carcinoma breast, and carcinoma lung, where patients presented with metastatic cutaneous lesions and positron-emission tomography-computed tomography (PET-CT) scan whole body helped in localizing the primary site. Whole-body fluorodeoxyglucose PET-CT scan helps in scanning whole body at once and detect occult primary and metastatic sites.

Placenta in intrauterine fetal demise (IUFD): a comprehensive study from a tertiary care hospital.

Background: Intrauterine fetal demise (IUFD) is an unpredictable and challenging obstetric complication. Its etiology is multifactorial with more than 60% attributed to the placental cause. The present study was done with a primary objective of understanding the placental lesions underlying IUFD. Methods: In this retrospective observational study, IUFD cases (>22 weeks) between January 2012 and September 2015 were collected from pathology database. The clinical details with ultrasound findings were collected from mother's charts. The lesions were classified into (A) maternal vascular malperfusion (MVM) including retroplacental hematomas, (B) fetal vascular malperfusion (FVM), (C) inflammatory lesions, and (D) idiopathic. The contributor to fetal death was classified as direct, major, minor, unlikely, or unknown. Placental findings of fetal hypoxia were recorded. Results: The study included 100 cases of IUFD. The mean maternal age was 26 years (18-36 years). Primipara were 46. There were 65 early preterm (PT) (<34 weeks), 20 late PT (34 weeks to <37 weeks) and 15 term (>37 weeks) IUFD. The mean gestation age was 30 weeks. The ratio of male:female fetuses was 1:1.7. Relevant obstetric complications included preeclampsia (n = 39), intrauterine growth restriction (IUGR) (n = 7), pre-gestational diabetes (n = 7), bad obstetric history (n = 6), oligohydramnios (n = 5). The mean placental weight was 256 g. Maternal vascular malperfusion had the highest incidence (30%), followed by combined maternal and FVM (10%). Exclusive inflammatory lesions and FVM were seen in 12 and 6%, respectively. No cause was identified in 18%. Direct contributor to IUFD was identified in 51 cases and major, minor, unlikely contribution in 21, 11 and nine cases, respectively. In nine cases, it was unknown. Lesions indicating fetal hypoxia were noted in 35 cases. In both early and late PT, MVM featured more commonly (23 and 5%). In term placentas, the most common cause was idiopathic. Conclusions: Lesions of MVM were the most common cause of IUFD and served as a direct contributor to fetal demise.