The Alk receptor tyrosine kinase regulates Sparkly, a novel activity regulating neuropeptide precursor in the Drosophila central nervous system.

Numerous roles for the Alk receptor tyrosine kinase have been described in Drosophila, including functions in the central nervous system (CNS), however the molecular details are poorly understood. To gain mechanistic insight, we employed Targeted DamID (TaDa) transcriptional profiling to identify targets of Alk signaling in the larval CNS. TaDa was employed in larval CNS tissues, while genetically manipulating Alk signaling output. The resulting TaDa data were analyzed together with larval CNS scRNA-seq datasets performed under similar conditions, identifying a role for Alk in the transcriptional regulation of neuroendocrine gene expression. Further integration with bulk and scRNA-seq datasets from larval brains in which Alk signaling was manipulated identified a previously uncharacterized Drosophila neuropeptide precursor encoded by CG4577 as an Alk signaling transcriptional target. CG4577, which we named Sparkly (Spar), is expressed in a subset of Alk-positive neuroendocrine cells in the developing larval CNS, including circadian clock neurons. In agreement with our TaDa analysis, overexpression of the Drosophila Alk ligand Jeb resulted in increased levels of Spar protein in the larval CNS. We show that Spar protein is expressed in circadian (clock) neurons, and flies lacking Spar exhibit defects in sleep and circadian activity control. In summary, we report a novel activity regulating neuropeptide precursor gene that is regulated by Alk signaling in the Drosophila CNS.

Extremity Injuries in Pillion Riders of Fatal 2-Wheeler Road Traffic Accidents: An Autopsy-Based Study.

ABSTRACT: The extremities are the important anatomical regions that are disproportionately injured during road traffic accidents in poor and medium-income countries. The data regarding extremity injuries, particularly in pillion passengers, are minimal globally. We analyzed the pattern of extremity injuries and their association with various parameters such as collision type, seating position, road type, and type of vehicle. This was a hospital-based cross-sectional study. The pillion riders of fatal motorized 2-wheeler road accident cases that were subjected to autopsy were studied over a period of 2 years. We analyzed 73 cases of pillion riders. Females (60%) were the most commonly affected group. The upper extremities were injured in 55 cases (75.3%) and the lower extremities in 49 cases (67.1%). The shoulder (41%), hand (31.3%), and elbow (23.2%) were the predominantly injured anatomical regions of the upper limb. In the lower extremity, knee (41%) and foot (21.9%) were mostly affected. Forty-nine (67.1%) and 48 (65.8%) cases had abrasions in the upper limbs and lower limbs, respectively, followed by lacerations and contusions. Upper extremity injuries are more common compared with lower extremity. The majority of the pillion riders were seated in a side-saddle position, and self-fall from the bike without any collision with other vehicles was the most common mode of injury. Thus, appropriate seating posture with safety gear can be used to reduce extremity injuries.

Medical negligence in cases decided by the National Consumer Disputes Redressal Commission: A five-year retrospective review.

BACKGROUND: There has been a gradual increase in disputes between doctors and patients in the healthcare system over the years. The aim of this review was to determine the speciality-wise prevalence of medical negligence in cases decided by the National Consumer Disputes Redressal Commission (NCDRC) and the factors responsible for it. METHODS: A total of 253 cases of medical negligence decided by the NCDRC from 2015 to 2019 were reviewed and categorised with respect to the number of cases compensated, the speciality involved, the compensation payout for the specialities involved, and the nature of the error leading to negligence. RESULTS: Among the cases analysed, negligence was identified in 135(53%) cases. Of these, the incidence of negligence was highest in surgery [37(27%)], followed by obstetrics and gynaecology (OBG) [29(21%)]. The highest compensation payouts were Rs 1.38 crore and Rs 1.1 crore in the paediatrics and OBG specialties, respectively. The common errors were lack of skill/care in the treatment of the patient [62(36%)] and failure to maintain accurate medical records [38 (22%)]. CONCLUSION: The study of adverse events in healthcare practice can improve the quality of patient care, and steps can be taken to reduce such events. Many adverse events are preventable by improving the skill/care in treatment and meticulous record keeping.

DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as an Alk target in the Drosophila visceral mesoderm.

Development of the Drosophila visceral muscle depends on Anaplastic Lymphoma Kinase (Alk) receptor tyrosine kinase (RTK) signaling, which specifies founder cells (FCs) in the circular visceral mesoderm (VM). Although Alk activation by its ligand Jelly Belly (Jeb) is well characterized, few target molecules have been identified. Here, we used targeted DamID (TaDa) to identify Alk targets in embryos overexpressing Jeb versus embryos with abrogated Alk activity, revealing differentially expressed genes, including the Snail/Scratch family transcription factor Kahuli (Kah). We confirmed Kah mRNA and protein expression in the VM, and identified midgut constriction defects in Kah mutants similar to those of pointed (pnt). ChIP and RNA-Seq data analysis defined a Kah target-binding site similar to that of Snail, and identified a set of common target genes putatively regulated by Kah and Pnt during midgut constriction. Taken together, we report a rich dataset of Alk-responsive loci in the embryonic VM and functionally characterize the role of Kah in the regulation of embryonic midgut morphogenesis.

In vivo Profiling of the Alk Proximitome in the Developing Drosophila Brain.

Anaplastic lymphoma kinase (Alk) is an evolutionary conserved receptor tyrosine kinase belonging to the insulin receptor superfamily. In addition to its well-studied role in cancer, numerous studies have revealed that Alk signaling is associated with a variety of complex traits such as: regulation of growth and metabolism, hibernation, regulation of neurotransmitters, synaptic coupling, axon targeting, decision making, memory formation and learning, alcohol use disorder, as well as steroid hormone metabolism. In this study, we used BioID-based in vivo proximity labeling to identify molecules that interact with Alk in the Drosophila central nervous system (CNS). To do this, we used CRISPR/Cas9 induced homology-directed repair (HDR) to modify the endogenous Alk locus to produce first and next generation Alk::BioID chimeras. This approach allowed identification of Alk proximitomes under physiological conditions and without overexpression. Our results show that the next generation of BioID proteins (TurboID and miniTurbo) outperform the first generation BirA* fusion in terms of labeling speed and efficiency. LC-MS3-based BioID screening of AlkTurboID and AlkminiTurbo larval brains revealed an extensive neuronal Alk proximitome identifying numerous potential components of Alk signaling complexes. Validation of Alk proximitome candidates further revealed co-expression of Stardust (Sdt), Discs large 1 (Dlg1), Syntaxin (Syx) and Rugose (Rg) with Alk in the CNS and identified the protein-tyrosine-phosphatase Corkscrew (Csw) as a modulator of Alk signaling.

Identification of the Wallenda JNKKK as an Alk suppressor reveals increased competitiveness of Alk-expressing cells.

Anaplastic lymphoma kinase (Alk) is a receptor tyrosine kinase of the insulin receptor super-family that functions as oncogenic driver in a range of human cancers such as neuroblastoma. In order to investigate mechanisms underlying Alk oncogenic signaling, we conducted a genetic suppressor screen in Drosophila melanogaster. Our screen identified multiple loci important for Alk signaling, including members of Ras/Raf/ERK-, Pi3K-, and STAT-pathways as well as tailless (tll) and foxo whose orthologues NR2E1/TLX and FOXO3 are transcription factors implicated in human neuroblastoma. Many of the identified suppressors were also able to modulate signaling output from activated oncogenic variants of human ALK, suggesting that our screen identified targets likely relevant in a wide range of contexts. Interestingly, two misexpression alleles of wallenda (wnd, encoding a leucine zipper bearing kinase similar to human DLK and LZK) were among the strongest suppressors. We show that Alk expression leads to a growth advantage and induces cell death in surrounding cells. Our results suggest that Alk activity conveys a competitive advantage to cells, which can be reversed by over-expression of the JNK kinase kinase Wnd.

Clinical Profile and Autopsy Findings in Fatal Head Injuries.

AIMS: This study aims to correlate the autopsy findings with the clinical picture and imaging report in fatal head injury patients. SETTINGS AND DESIGN: A descriptive study conducted at tertiary care hospital in South India from July 2015 to December 2016. PATIENTS AND METHODS: All patients with head injuries who were admitted to our Emergency and Trauma Centre and underwent autopsy were included in the study. A structured pro forma was used for collecting information. Autopsy findings were considered as a gold standard to correlate with antemortem findings in fatal head injury. The data were analyzed with EpiData and OpenEpi statistical analyzing software. RESULTS: Of the 303 fatal head injury patients, a majority were males and age group between 21 and 40 years. Eighty-eight percent (267/303) of fatal head injuries were due to road traffic accidents. Twenty-five of the 303 patients reached our center within 1 h (golden hour) of trauma. Of the 303 fatal head injuries, 153 (50.5%) died within 24 h of reaching our center. The most common autopsy finding in this study was subarachnoid hemorrhage (SAH) (247/303, 81.3%). Diagnostic accuracy of Epi dural hemorrhage (EDH) antemortem had the highest value (98.35%). SAH had least diagnostic accuracy value (45.72). subdural hemorrhage (SDH) had highest sensitivity (57.02%). EDH had higher specificity (100%). Significant SDH, SAH, and brain contusions were not detected during antemortem evaluation. CONCLUSIONS: Our study revealed that among fatal head injury patients, half of them died within first 24 h after reaching to tertiary care center. Diagnostic accuracy to detect extradural hemorrhage antemortem had the highest value and SAH had least diagnostic accuracy value. Significant subdural hemorrhage, subarachniod hemorrhage, and brain contusion were not detected during antemortem evaluation. Expertise in interpretation of imaging, adequate clinical examination, proper documentation, and early resuscitation may reduce the chances of missed injuries in head injury patients.

dSTIM- and Ral/Exocyst-Mediated Synaptic Release from Pupal Dopaminergic Neurons Sustains Drosophila Flight.

Manifestation of appropriate behavior in adult animals requires developmental mechanisms that help in the formation of correctly wired neural circuits. Flight circuit development in Drosophila requires store-operated calcium entry (SOCE) through the STIM/Orai pathway. SOCE-associated flight deficits in adult Drosophila derive extensively from regulation of gene expression in pupal neurons, and one such SOCE-regulated gene encodes the small GTPase Ral. The cellular mechanism by which Ral helps in maturation of the flight circuit was not understood. Here, we show that knockdown of components of a Ral effector, the exocyst complex, in pupal neurons also leads to reduced flight bout durations, and this phenotype derives primarily from dopaminergic neurons. Importantly, synaptic release from pupal dopaminergic neurons is abrogated upon knockdown of dSTIM, Ral, or exocyst components. Ral overexpression restores the diminished synaptic release of dStim knockdown neurons as well as flight deficits associated with dSTIM knockdown in dopaminergic neurons. These results identify Ral-mediated vesicular release as an effector mechanism of neuronal SOCE in pupal dopaminergic neurons with functional consequences on flight behavior.

Occupational death due to bilateral carotid artery occlusion - a medico-legal case report.

Occupational deaths are work-related deaths and usually occur in the work place of an individual. A case of occupational death in a female factory worker has been described, where the saree she was wearing accidentally got entangled in the moving belt and caused ligature strangulation. She was unconscious throughout indicating bilateral occlusion of the carotids and died a day after the incident. The authors have described the correlation between the distribution of the injuries on the body and the manner a saree is worn. Investigation of the workplace revealed that the place where she was standing was located between two machines, and the space was inadequate for normal movement. Some suggestions have been offered which may decrease the incidences of occupational ligature strangulation due to entanglement of dress materials in the moving machineries.