A molecular switch for neuroprotective astrocyte reactivity.

The intrinsic mechanisms that regulate neurotoxic versus neuroprotective astrocyte phenotypes and their effects on central nervous system degeneration and repair remain poorly understood. Here we show that injured white matter astrocytes differentiate into two distinct C3-positive and C3-negative reactive populations, previously simplified as neurotoxic (A1) and neuroprotective (A2)1,2, which can be further subdivided into unique subpopulations defined by proliferation and differential gene expression signatures. We find the balance of neurotoxic versus neuroprotective astrocytes is regulated by discrete pools of compartmented cyclic adenosine monophosphate derived from soluble adenylyl cyclase and show that proliferating neuroprotective astrocytes inhibit microglial activation and downstream neurotoxic astrocyte differentiation to promote retinal ganglion cell survival. Finally, we report a new, therapeutically tractable viral vector to specifically target optic nerve head astrocytes and show that raising nuclear or depleting cytoplasmic cyclic AMP in reactive astrocytes inhibits deleterious microglial or macrophage cell activation and promotes retinal ganglion cell survival after optic nerve injury. Thus, soluble adenylyl cyclase and compartmented, nuclear- and cytoplasmic-localized cyclic adenosine monophosphate in reactive astrocytes act as a molecular switch for neuroprotective astrocyte reactivity that can be targeted to inhibit microglial activation and neurotoxic astrocyte differentiation to therapeutic effect. These data expand on and define new reactive astrocyte subtypes and represent a step towards the development of gliotherapeutics for the treatment of glaucoma and other optic neuropathies.

Reduced FOXF1 links unrepaired DNA damage to pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is a progressive disease in which pulmonary arterial (PA) endothelial cell (EC) dysfunction is associated with unrepaired DNA damage. BMPR2 is the most common genetic cause of PAH. We report that human PAEC with reduced BMPR2 have persistent DNA damage in room air after hypoxia (reoxygenation), as do mice with EC-specific deletion of Bmpr2 (EC-Bmpr2-/-) and persistent pulmonary hypertension. Similar findings are observed in PAEC with loss of the DNA damage sensor ATM, and in mice with Atm deleted in EC (EC-Atm-/-). Gene expression analysis of EC-Atm-/- and EC-Bmpr2-/- lung EC reveals reduced Foxf1, a transcription factor with selectivity for lung EC. Reducing FOXF1 in control PAEC induces DNA damage and impaired angiogenesis whereas transfection of FOXF1 in PAH PAEC repairs DNA damage and restores angiogenesis. Lung EC targeted delivery of Foxf1 to reoxygenated EC-Bmpr2-/- mice repairs DNA damage, induces angiogenesis and reverses pulmonary hypertension.

Immunofluorescent and molecular characterization of effusion tumor cells reveal cancer site-of-origin and disease-driving mutations.

BACKGROUND: Effective cancer treatment relies on precision diagnostics. In cytology, an accurate diagnosis facilitates the determination of proper therapeutics for patients with cancer. Previously, the authors developed a multiplexed immunofluorescent panel to detect epithelial malignancies from pleural effusion specimens. Their assay reliably distinguished effusion tumor cells (ETCs) from nonmalignant cells; however, it lacked the capacity to reveal specific cancer origin information. Furthermore, DNA profiling of ETCs revealed some, but not all, cancer-driver mutations. METHODS: The authors developed a new multiplex immunofluorescent panel that detected both malignancy and pulmonary origin by incorporating the thyroid transcription factor-1 (TTF-1) biomarker. Evaluation for TTF-1-positive ETCs (T-ETCs) was performed on 12 patient samples. T-ETCs and parallel ETCs from selected patients were collected and subjected to DNA profiling to identify pathogenic mutations. All samples were obtained with Institutional Review Board approval. RESULTS: Malignancy was detected in all samples. T-ETCs were identified in 9 of 10 patients who had clinically reported TTF-1 positivity (90% sensitivity and 100% specificity). Furthermore, DNA profiling of as few as five T-ETCs identified pathogenic mutations with equal or greater sensitivity compared with profiling of ETCs, both of which showed high concordance with clinical findings. CONCLUSIONS: The findings suggest that the immunofluorescent and molecular characterization of tumor cells from pleural effusion specimens can provide reliable diagnostic information, even with very few cells. The integration of site-specific biomarkers like TTF-1 into ETC analysis may facilitate better refined diagnosis and improve patient care.

Posterior Reversible Encephalopathy Syndrome After Epilepsy Surgery Alerted by Low-Processed Electroencephalography Levels: A Case Report.

The development of posterior reversible encephalopathy syndrome (PRES) in a patient undergoing epilepsy surgery without perioperative hypertension is uncommon. A young man having epilepsy surgery with normal blood pressures had an unexplained drop in his processed electroencephalogram (pEEG) levels intraoperatively. This alerted and prompted us to search for the cause. A postoperative electroencephalogram (EEG) confirmed a diffuse slowing of cortical waves. The intraoperative findings of pEEG, magnetic resonance imaging (MRI), and EEG postoperatively prompted a diagnosis of PRES. The patient was managed conservatively and had a full recovery. This case report highlights the role of brain electrical activity monitors in PRES.

Complete Resolution of Disease After Peptide Receptor Radionuclide Therapy in a Patient of Metastatic Insulinoma.

ABSTRACT: A 48-year-old man, a case of metastatic insulinoma, who failed transarterial chemoembolization of liver metastases underwent multiple cycles of peptide receptor radionuclide therapy with 177Lu-DOTATATE, following which a complete morphologic and metabolic response was demonstrated on 68Ga-DOTATATE PET/CT. Patient had a remarkable improvement in his quality of life as intractable hypoglycemic episodes resolved after treatment. Peptide receptor radionuclide therapy is a promising targeted radionuclide therapy in patients of metastatic insulinomas that can result in reduced tumor burden and improved quality of life, particularly those who fail the conventional treatment modalities as seen in the present case.

Comparison of Multiparametric Magnetic Resonance Imaging and Gallium-68 Prostate-Specific Membrane Antigen Positron Emission Tomography/Computed Tomography for Detecting Carcinoma Prostate in Patients with Serum Prostate-Specific Antigen between 4 and 20 ng/ml.

INTRODUCTION: We carried out this study to compare the diagnostic accuracy of multiparametric magnetic resonance imaging (mpMRI) and gallium-68 prostate-specific membrane antigen positron emission tomography/computed tomography (Ga-68 PSMA PET/CT) to detect prostatic carcinoma in patients with serum prostate-specific antigen (PSA) between 4 and 20 ng/ml in prebiopsy setting. MATERIALS AND METHODS: This prospective study evaluated men with serum PSA values between 4 and 20 ng/ml. All patients underwent mpMRI and Ga-68 PSMA PET/CT, followed by 12-core transrectal ultrasonography (TRUS)-guided biopsy to detect prostatic carcinoma. The diagnostic accuracy of mpMRI and PSMA PET/CT scan was compared with histopathological findings. RESULTS: There were thirty patients included in the study with a median age of 73 years (age range: 69-79 years). The median total serum PSA was 8.0 ng/ml (5.0-19.9 ng/ml). Of these, 18 had an identifiable lesion on imaging and had histopathological findings suggestive of carcinoma prostate. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of mpMRI were 100%, 92.30%, 94.73%, and 100%, respectively, and that of PSMA PET scan were 94.44%, 100%, 100%, and 92.31%, respectively. The diagnostic accuracy of both was 96.67%. CONCLUSION: PSMA PET scan showed higher PPV and specificity while mpMRI showed higher sensitivity and NPV. The accuracy in predicting presence of carcinoma was the same for both. PSMA PET showed higher specificity and PPV and predicted the subsequent need of biopsy. In our study, the NPV of PET, though good, was lower than mpMRI. Prospective trials with larger sample size are needed. In combination, PET/MRI may achieve greater accuracy and may serve as investigation of choice.

A reversed phase HPLC-UV method for the simultaneous determination of residual formaldehyde and Triton X-100 in vaccine products.

Many of the inactivated viral vaccines for human and animal use are manufactured using formaldehyde as an inactivating agent. Apart from formaldehyde, Triton X-100 is also one of the chemicals commonly used in viral vaccine manufacturing. Triton X-100 is typically used to extract the cell-associated viruses and / or components during manufacturing process. The concentration of formaldehyde and Triton X-100 in the final bulks are also reduced during vaccine purification process. Here we report a simple RP-HPLC-UV based method for the quantification of residual Triton X-100 and formaldehyde as process impurities in viral vaccines. This method is also adopted for the residual impurity determination of either formaldehyde or Triton X-100 in other non-viral vaccines, multivalent as well as sub-unit vaccines, such as liquid pentavalent, includes TT, DT, Hepatitis B (rDNA) and Haemophilus type b conjugate vaccine (adsorbed). This method is rapid and can quantify both Triton X-100 and formaldehyde in a single preparation with improved peak asymmetry. This new assay has a linearity range starting from 0.0625 to 1 µg/mL for formaldehyde and 0.625-10 µg/mL for Triton X-100. This method would be very useful for viral vaccine manufacturing and release.

Ga-68 DOTATATE Positron Emission Tomography/Computed Tomography in a Rare Case of Esthesioneuroblastoma.

We describe the Ga-68 DOTATATE positron emission tomography/computed tomography (PET/CT) findings of a 51-year-old man, operated for right esthesioneuroblastoma. Postoperative Ga-68 DOTATATE PET/CT revealed focal uptake anterior to sphenoid ostium on the right paramedian side, suspicious for residual disease. Magnetic resonance imaging showed an enhancing lesion in posterosuperior nasal cavity on the right side extending into the right sphenoid sinus. He underwent re-surgery and adjuvant chemoradiotherapy. The histopathology revealed residual olfactory neuroblastoma. The follow-up Ga-68 DOTATATE PET/CT was negative. This case emphasizes the role of Ga-68 DOTATATE PET/CT in the management, especially in residual or recurrent disease and potential radiotheranostics for these rare tumors.

Utility of Blood as the Clinical Specimen for the Molecular Diagnosis of Post-Kala-Azar Dermal Leishmaniasis.

The countries in the Indian subcontinent have reported a dramatic decline in visceral leishmaniasis (VL) cases. However, the presence of the parasite reservoir in the form of post-kala-azar dermal leishmaniasis (PKDL), a dermal sequel of VL, is a hurdle in attaining VL elimination. Presently employed clinical specimens for the diagnosis of PKDL include skin biopsy specimens and slit skin smears. In this study, the use of blood as a clinical specimen was investigated in different manifestations of PKDL in India. This is a bicentric study (National Institute of Pathology, Indian Council of Medical Research [ICMR], New Delhi, and Institute of Medical Sciences [IMS], Banaras Hindu University, Varanasi), with 215 participants (120 PKDL patients and 95 controls). Highly sensitive quantitative real-time PCR (Q-PCR) and field-deployable loop-mediated isothermal amplification (LAMP) were employed using blood samples for diagnosis. Promising sensitivities of 77.50% (95% confidence interval [CI], 69.24 to 84.05%) for Q-PCR and 70.83% (95% CI, 62.16 to 78.22%) for LAMP were obtained for the diagnosis of PKDL. Further, enhanced sensitivities of 83.33% (95% CI, 71.28 to 90.98%) and 77.78% (95% CI, 65.06 to 86.80%) for Q-PCR and LAMP, respectively, were recorded for the detection of macular cases. The study revealed an inverse correlation between the parasite load estimated in slit and blood samples, thereby favoring the use of blood for the diagnosis of the macular variant, which may be missed due to scant parasite loads in the slit. This study is the first to propose the promising potential of blood as a clinical specimen for accurate diagnosis of PKDL, which would aid in fast-tracking VL elimination.

Diagnostic Dilemma of Cutaneous Tuberculosis: Opening of the Pandora's Box.

ABSTRACT: Tuberculosis in all forms, that is, pulmonary (PTB) or extrapulmonary (EPTB), is a universal health problem. Cutaneous tuberculosis (CTb) remains one of the least studied and often under-reported variants of EPTB because of its wide and protean clinical presentation. The diagnosis of CTb remains challenging because of lack of sensitive and specific investigations for its diagnosis. The sensitivity of some of the traditional tests is low because of low concentration of mycobacteria in paucibacillary lesions. Besides it is difficult to distinguish between M. tuberculosis (MTb) and other mycobacterial species in skin biopsies morphologically. Molecular methods may target either MTb DNA or RNA, and serve as promising tools in the diagnosis of various forms of CTb, with high sensitivity and rapidity. This review is focused on diagnostic challenges of CTb and to discuss various methods and newer technologies for diagnosing CTb. This will help the dermatologists and dermatopathologists to elucidate and accurately diagnose CTb from other infectious granulomatous dermatitis for appropriate timely treatment of the patient.

Prostate-Specific Membrane Antigen Expression in Patients With Differentiated Thyroid Cancer With Thyroglobulin Elevation and Negative Iodine Scintigraphy Using 68Ga-PSMA-HBED-CC PET/CT.

PURPOSE OF THE REPORT: Prostate-specific membrane antigen (PSMA) is a member of superfamily of zinc-dependent exopeptidases that is robustly expressed in prostate cancer cells and nonprostatic solid tumor neovasculature including microvessels of thyroid tumors. Its expression in differentiated thyroid cancer (DTC) has been confirmed in many recent studies, but systematic studies exploring PSMA expression in patients with DTC with thyroglobulin elevation and negative iodine scintigraphy (TENIS) are lacking. The aim of the present study was to evaluate the role of PSMA scan in TENIS patients with DTC. METHODS: Nine consecutive patients with DTC with proven TENIS syndrome (6 men and 3 women with age range 29-68 years and mean age of 48 years) underwent 18F-FDG PET/CT as per the institution protocol. Thereafter, they were subjected to 68Ga-PSMA-HBED-CC PET/CT as per the institution protocol within a week of FDG PET imaging. Prostate-specific membrane antigen expression (SUVmax) in the lesions was compared with 18F-FDG PET and CT scan findings. RESULTS: In 5 of 9 patients with TENIS, the metastatic lesions showed PSMA expression. A total of 14 lesions were seen on the CT scan. Prostate-specific membrane antigen PET detected 9 of 14 lesions (64.28%) (SUVmax ranging from 10.1 to 45.67; median SUVmax of 16.31), whereas FDG PET was positive in 11 of 14 lesions (78.57%). The lesions that showed PSMA uptake was localized to bones (5 of 9) and lungs (4 of 9). Two lesions that were localized to iliac crest and acetabulum were missed on FDG PET but were seen on CT and PSMA PET scan. CONCLUSIONS: The results of this pilot study indicate that 68Ga-HBED-CC-PSMA PET/CT demonstrates PSMA expression in TENIS patients with lesions being localized to the bones and lungs. 68Ga-PSMA PET/CT could be useful for the identification of TENIS patients who might benefit from PSMA-targeted radionuclide therapy.

68Ga-PSMA-HBED-CC PET/CT Findings in a Patient of Polyostotic Fibrous Dysplasia.

ABSTRACT: A 43-year-old man diagnosed with fibrous dysplasia with McCune-Albright syndrome was subjected to 18F-fluoride bone scan and 68Ga-PSMA-HBED-CC PET/CT as per the institution protocol. 18F-bone scan revealed extensive involvement of axial and appendicular skeleton confirming polyostotic fibrous dysplasia. 68Ga-PSMA PET/CT showed increased tracer uptake in corresponding lesions of fibrous dysplasia. PSMA uptake in fibrous dysplasia lesions has been rarely described with literature evidence being limited to anecdotal case reports. Nevertheless, due to increasing use of PSMA PET/CT, one should be aware of this false-positive finding to avoid misinterpretation of the scans.

Cutaneous tuberculosis of the pinna.

BACKGROUND: Cutaneous tuberculosis (CTB) rarely involves the ear as the primary site, but while diagnosing and treating ear infections, it should be considered a differential diagnosis in a tropical country such as India. The present study reports the incidence and clinical presentation of auricular tuberculosis (TB) in a tertiary care hospital in New Delhi. METHODS: A retrospective, observational study was conducted from 2005 to 2019 whereby all cases of CTB confirmed by biopsy were retrieved from the database. The demographic details, clinical details, Mantoux results, and photographs were extracted and studied. The data were entered into MS Excel and analyzed. RESULTS: In a retrospective analysis of 886 cases of CTB over a period of 15 years, we found 20 cases (2.26%) of ear involvement (1 case with bilateral involvement). The median age of the patients with ear involvement was 29 years with 42.11% men and 57.89% women. Morphological variants seen over pinna were predominantly classic plaque type (31.58%) and nodular (31.58%), with few ulcerative (21.05%) and tumoral forms (15.79%). CTB of the pinna showed predominant involvement of either helix or ear lobule (7 cases each). All cases were strongly positive to tuberculin and showed response to the empirical antitubercular treatment. CONCLUSION: CTB can exclusively affect the pinna in varied presentations. The ear lobules and the helix are the usual sites of affection. It is rare for both ears to be affected with CTB, unlike bacilliferous leprosy. Regression following institution of antitubercular treatment is a reasonable way to confirm CTB.

18F-FDG PET/CT in a Rare Case of Poland Syndrome and Gastric Cancer.

ABSTRACT: Poland syndrome is a rare congenital anomaly characterized by unilateral aplasia of the sternoclavicular head of pectoralis major muscle with varying degree of same side upper limb anomalies. A 44-year-old man, with a case of adenocarcinoma of stomach, whose CECT chest revealed complete absence of pectoralis major and minor muscles on the left side, was diagnosed with Poland syndrome without presence of typical ipsilateral limb anomalies. Follow-up PET/CT revealed metabolically active recurrent disease with typical findings of Poland syndrome. It is important to be aware of oncologic association in a patient of Poland syndrome as highlighted in the present case.

A clinical, dermoscopic, histopathological and immunohistochemical study of melasma and facial pigmentary demarcation lines in the skin of color.

Melasma and facial pigmentary demarcation lines (FPDL) are common causes of patterned facial pigmentation that may mimic each other. There is a paucity of studies investigating these two conditions. The objective of this study was to make a detailed comparative analysis of these disorders. A clinical, dermoscopic, histopathological and immunohistochemical analysis of lesional and perilesional skin was conducted in 20 patients each of melasma and FDPL. The most common morphological patterns were centrofacial in melasma and W-shaped pattern in FPDL. Dermoscopy in melasma revealed similar patterns in lesional and perilesional skin, whereas FPDL did not. Histopathology of melasma revealed increased melanin in the suprabasal and basal layers (100%), melanophages in the upper dermis and solar elastosis (65%) in contrast to FPDL, wherein increased basilar melanin (75%) and dermal melanophages were the key findings. Expression of vascular endothelial growth factor and stem cell factor was slightly increased in lesional melasma skin, but not in FPDL. The study was limited by its small sample size and immunohistochemistry carried out in a few patients. Melasma and FPDL, although similar in presentation, are distinct entities. Dermoscopy, histology and immunohistochemistry reveal subtle differences.

One decade of 'Bench-to-Bedside' peptide receptor radionuclide therapy with indigenous [177Lu]Lu-DOTATATE obtained through 'Direct' neutron activation route: lessons learnt including practice evolution in an Indian setting.

The present treatise chronicles one decade of experience pertaining to clinical PRRT services in a large-volume tertiary cancer care centre in India delivering over 4,000 therapies, an exemplar of successful PRRT programme employing indigenous 177Lutetium production and resources. For the purpose of systematic discussion, we have sub-divided the communication into 3 specific parts: (a) Radiopharmaceutical aspects that describes 177Lutetium production through 'Direct' Neutron Activation Route and the subsequent radiolabeling procedures, (b) The specific clinical nuances and finer learning points (apart from the routine standard procedure) based upon clinical experience and how it has undergone practice evolution in our setting and (c) Dosimetry results with this indigenous product and radiation safety/health physics aspects involved in PRRT services. Initiated in 2010 at our centre, the PRRT programme is a perfect example of affordable quality health care delivery, with indigenous production of the radionuclide (177Lu) in the reactor and subsequent radiolabeling of the radiopharmaceutical ([177Lu]Lu-DOTATATE) at the hospital radiopharmacy unit of the centre, which enabled catering to the needs of a large number of patients of progressive, metastatic and advanced Neuroendocrine Neoplasms (NENs) and related malignancies.

Elevated IL-6R on CD4+ T cells promotes IL-6 driven Th17 cell responses in patients with T1R leprosy reactions.

Th17 cells play vital role during pathogenesis of leprosy reactions. Previously, we have reported that IL-23 is involved in Th17 cells differentiation. Subsequently, our group also showed that IL-6 induces Th17 cell differentiation along with TGF-ß in leprosy reactions. Here, we next asked the question that whether IL-6 or IL-23 induced Th17 cells are different in nature? In this study, Type 1 Reactions (T1R) showed significantly (p < 0.001) higher percentage of IL-17A producing CD4+IL6R+ T cells as compared to non-reaction (NR) patients. Furthermore, recombinant IL-6, IL-23 and TGF-ß promoted IL-17A secretion by CD4+IL6R+ T cells. Subsequently, IL-6R and IL-23R blocking experiments showed significantly (p < 0.002) down regulated IL-17A in T1R reaction as compared to NR leprosy patients. The present study for the first time establishes that pathogenic Th17 cells produce IL-17 in an IL-6 dependent manner in leprosy T1R reactions. Thus, present approaches that specifically target Th17 cells and/or the cytokines that promote their development, such as IL-6, TGF-ß and IL-23A may provide more focused treatment strategies for the management of Mycobacterium leprae and its reactions.