COPII cage assembly factor Sec13 integrates information flow regulating endomembrane function in response to human variation.

How information flow is coordinated for managing transit of 1/3 of the genome through endomembrane pathways by the coat complex II (COPII) system in response to human variation remains an enigma. By examining the interactome of the COPII cage-assembly component Sec13, we show that it is simultaneously associated with multiple protein complexes that facilitate different features of a continuous program of chromatin organization, transcription, translation, trafficking, and degradation steps that are differentially sensitive to Sec13 levels. For the trafficking step, and unlike other COPII components, reduction of Sec13 expression decreased the ubiquitination and degradation of wild-type (WT) and F508del variant cargo protein cystic fibrosis transmembrane conductance regulator (CFTR) leading to a striking increase in fold stability suggesting that the events differentiating export from degradation are critically dependent on COPII cage assembly at the ER Golgi intermediate compartment (ERGIC) associated recycling and degradation step linked to COPI exchange. Given Sec13's multiple roles in protein complex assemblies that change in response to its expression, we suggest that Sec13 serves as an unanticipated master regulator coordinating information flow from the genome to the proteome to facilitate spatial covariant features initiating and maintaining design and function of membrane architecture in response to human variation.

Autosomal recessive non-syndromic hearing loss genes in Pakistan during the previous three decades.

Hearing loss is a clinically and genetically heterogeneous disorder, with over 148 genes and 170 loci associated with its pathogenesis. The spectrum and frequency of causal variants vary across different genetic ancestries and are more prevalent in populations that practice consanguineous marriages. Pakistan has a rich history of autosomal recessive gene discovery related to non-syndromic hearing loss. Since the first linkage analysis with a Pakistani family that led to the mapping of the DFNB1 locus on chromosome 13, 51 genes associated with this disorder have been identified in this population. Among these, 13 of the most prevalent genes, namely CDH23, CIB2, CLDN14, GJB2, HGF, MARVELD2, MYO7A, MYO15A, MSRB3, OTOF, SLC26A4, TMC1 and TMPRSS3, account for more than half of all cases of profound hearing loss, while the prevalence of other genes is less than 2% individually. In this review, we discuss the most common autosomal recessive non-syndromic hearing loss genes in Pakistani individuals as well as the genetic mapping and sequencing approaches used to discover them. Furthermore, we identified enriched gene ontology terms and common pathways involved in these 51 autosomal recessive non-syndromic hearing loss genes to gain a better understanding of the underlying mechanisms. Establishing a molecular understanding of the disorder may aid in reducing its future prevalence by enabling timely diagnostics and genetic counselling, leading to more effective clinical management and treatments of hearing loss.

Brassinosteroids as promoters of seedling growth and antioxidant activity under heavy metal zinc stress in mung bean (Vigna radiata L.).

The escalation of harmful pollutants, including heavy metals, due to industrialization and urbanization has become a global concern. To mitigate the negative impacts of heavy metal stress on germination and early plant development, growth regulators have been employed. This study aimed to evaluate the response of mung bean (Vigna radiata L.) to zinc stress in the presence of brassinosteroids, focusing on seedling growth and antioxidant potential. Mung bean seedlings were treated with three concentrations of 24-epibrassinolide (EBL) (0.1, 0.2, and 0.4 PPM) with or without zinc. Results demonstrated that the application of brassinosteroids, combined with zinc stress, significantly enhanced germination percentage (about 47.06, 63.64, and 120%), speed of germination (about 39.13, 50, and 100%), seedling growth (about 38% in case of treatment combined 0.4 PPM 24-EBL and 1.5 mM ZnSO4) and seedling vigor index (204% in case of treatment combined 0.4 PPM 24-EBL and 1.5 mM ZnSO4) compared to zinc-treated seedlings alone after 24 h. The activities of antioxidative enzymes (catalase, ascorbate peroxidase, polyphenol oxidase, and peroxidase) and total soluble protein content decreased, while lipid peroxidation and proline content exhibited a significant increase (p ≤ 0.05) when compared to the control. However, the negative effects induced by heavy metal stress on these parameters were significantly mitigated by EBL application. Notably, the most effective concentration of EBL in overcoming zinc stress was found to be 0.4 PPM. These findings underscore the potential of exogenously applied brassinosteroids as a valuable tool in phytoremediation projects by ameliorating heavy metal stress.

Customised Implant for Temporomandibular Joint: New Technique to Design and Stress Analysis to Balance the Loading at Both Ends.

The temporomandibular joint (TMJ) is a critical joint for the opening and closing of the mouth. The generation of customised TMJs according to individuals' dental anatomy is needed. Currently, the implants available on the market lack consideration of the patient's dental anatomy. This leads to the creation of an imbalance in the reaction forces on both ends of the TMJ. This requires a slight structural change in the design parameters to give a solution. The purpose of this study is to propose a new design that includes the geometry and materials for a TMJ implant. Stress analysis was carried out on the TMJ to balance the reaction forces at both TMJ ends. A static analysis was performed using ANSYS Workbench, to compare the results of two customised designs of TMJ implants, in order to better balance the reaction forces at both ends. The model in the study showed that the reaction forces for both the patient-specific TMJ implants were nearly balanced. The reaction forces were better balanced, and almost equivalent to the intact conditions. The stresses in the mandible were more uniformly distributed in the customised design of the TMJ implant. The two types of design showed that the custom design took up less space in the patient's region of surgery, making it a better option compared to a stock TMJ implant. The custom implant would allow faster patient rehabilitation, as the reaction forces would be close to those in intact conditions.

A cryptic microdeletion del(12)(p11.21p11.23) within an unbalanced translocation t(7;12)(q21.13;q23.1) implicates new candidate loci for intellectual disability and Kallmann syndrome.

In a patient diagnosed with both Kallmann syndrome (KS) and intellectual disability (ID), who carried an apparently balanced translocation t(7;12)(q22;q24)dn, array comparative genomic hybridization (aCGH) disclosed a cryptic heterozygous 4.7 Mb deletion del(12)(p11.21p11.23), unrelated to the translocation breakpoint. This novel discovery prompted us to consider the possibility that the combination of KS and neurological disorder in this patient could be attributed to gene(s) within this specific deletion at 12p11.21-12p11.23, rather than disrupted or dysregulated genes at the translocation breakpoints. To further support this hypothesis, we expanded our study by screening five candidate genes at both breakpoints of the chromosomal translocation in a cohort of 48 KS patients. However, no mutations were found, thus reinforcing our supposition. In order to delve deeper into the characterization of the 12p11.21-12p11.23 region, we enlisted six additional patients with small copy number variations (CNVs) and analyzed eight individuals carrying small CNVs in this region from the DECIPHER database. Our investigation utilized a combination of complementary approaches. Firstly, we conducted a comprehensive phenotypic-genotypic comparison of reported CNV cases. Additionally, we reviewed knockout animal models that exhibit phenotypic similarities to human conditions. Moreover, we analyzed reported variants in candidate genes and explored their association with corresponding phenotypes. Lastly, we examined the interacting genes associated with these phenotypes to gain further insights. As a result, we identified a dozen candidate genes: TSPAN11 as a potential KS candidate gene, TM7SF3, STK38L, ARNTL2, ERGIC2, TMTC1, DENND5B, and ETFBKMT as candidate genes for the neurodevelopmental disorder, and INTS13, REP15, PPFIBP1, and FAR2 as candidate genes for KS with ID. Notably, the high-level expression pattern of these genes in relevant human tissues further supported their candidacy. Based on our findings, we propose that dosage alterations of these candidate genes may contribute to sexual and/or cognitive impairments observed in patients with KS and/or ID. However, the confirmation of their causal roles necessitates further identification of point mutations in these candidate genes through next-generation sequencing.

Button-Hole Disco-Osteotomy Drainage-A Novel Approach for High-Risk Patients with Cervical Abscess.

Spinal epidural abscess (SEA) is a very serious infection of the central nervous system (CNS). It is of very low incidence with a peak age in the geriatric age group. Immunocompromised patients are more susceptible to SEA. It can present with significant neurological deficits, which can be permanent if not identified and treated promptly. In this case report, a 75-year-old immunocompromised patient presented with progressive spastic quadriparesis and septicemia. He was diagnosed with a case of cervical spinal epidural abscess with underlying cord compression. Anterior retropharyngeal approach and button-hole disco-osteotomy of C5-C6 was performed and the cervical SEA was drained, followed by antibiotic saline irrigation (cranially and caudally) was done, total duration of surgery was 70 min. At the time of discharge (7th postoperative day), the patient improved neurologically and sepsis had resolved.

Currarino Syndrome Presenting in Adulthood: A Rare Case.

Currarino syndrome or Currarino triad is a complex condition consisting of congenital anomalies. The triad consists of anterior sacral mass (meningocele, teratoma or dermoid/epidermoid cyst), sacral bone defect (hypoplasia, agenesis ), anorectal malformation/congenital anorectal stenosis. This condition is named after Dr Guido Currarino, an Italian-American paediatric radiologist, who first described it in 1975. This needs a multidisciplinary treatment approach. We describe a case of successfully managed young adult with Currarino syndrome. The latest artificial intelligence tool, Chat Generative Pre-Trained Transformer (ChatGPT), helped to write this case report.

A circuit for secretion-coupled cellular autonomy in multicellular eukaryotic cells.

Cancers represent complex autonomous systems, displaying self-sufficiency in growth signaling. Autonomous growth is fueled by a cancer cell's ability to "secrete-and-sense" growth factors (GFs): a poorly understood phenomenon. Using an integrated computational and experimental approach, here we dissect the impact of a feedback-coupled GTPase circuit within the secretory pathway that imparts secretion-coupled autonomy. The circuit is assembled when the Ras-superfamily monomeric GTPase Arf1, and the heterotrimeric GTPase Giαßγ and their corresponding GAPs and GEFs are coupled by GIV/Girdin, a protein that is known to fuel aggressive traits in diverse cancers. One forward and two key negative feedback loops within the circuit create closed-loop control, allow the two GTPases to coregulate each other, and convert the expected switch-like behavior of Arf1-dependent secretion into an unexpected dose-response alignment behavior of sensing and secretion. Such behavior translates into cell survival that is self-sustained by stimulus-proportionate secretion. Proteomic studies and protein-protein interaction network analyses pinpoint GFs (e.g., the epidermal GF) as key stimuli for such self-sustenance. Findings highlight how the enhanced coupling of two biological switches in cancer cells is critical for multiscale feedback control to achieve secretion-coupled autonomy of growth factors.

Structural and Biophysical Characterization of Stable Alpha-Synuclein Oligomers.

The aggregation of α-synuclein (α-syn) into neurotoxic oligomers and fibrils is an important pathogenic feature of synucleinopatheis, including Parkinson's disease (PD). A further characteristic of PD is the oxidative stress that results in the formation of aldehydes by lipid peroxidation. It has been reported that the brains of deceased patients with PD contain high levels of protein oligomers that are cross-linked to these aldehydes. Increasing evidence also suggests that prefibrillar oligomeric species are more toxic than the mature amyloid fibrils. However, due to the heterogenous and metastable nature, characterization of the α-syn oligomeric species has been challenging. Here, we generated and characterized distinct α-syn oligomers in vitro in the presence of DA and lipid peroxidation products 4-hydroxy-2-nonenal (HNE) and 4-oxo-2-nonenal (ONE). HNE and ONE oligomer were stable towards the treatment with SDS, urea, and temperature. The secondary structure analysis revealed that only HNE and ONE oligomers contain ß-sheet content. In the seeding assay, both DA and ONE oligomers significantly accelerated the aggregation. Furthermore, all oligomeric preparations were found to seed the aggregation of α-syn monomers in vitro and found to be cytotoxic when added to SH-SY5Y cells. Finally, both HNE and ONE α-syn oligomers can be used as a calibrator in an α-syn oligomers-specific ELISA.

Intracardiac Repair in Late Adolescent and Adult Tetralogy of Fallot — Early and Midterm Results from a Tertiary Care Centre

Abstract Introduction: In developing countries like India, it is common for late presentation of Tetralogy of Fallot (TOF) patients to a hospital as compared to that of developed countries. The objective of this study is to analyze the surgical outcome of TOF patients with age > 15 years. Methods: This is a retrospective descriptive study of the surgical outcomes of 45 adult patients undergoing correction for TOF. Epidemiology, symptomology, and preoperative evaluation were performed. Results: Most of the patients were male (33 [73%]). The median age was 21 years. A total of 42 (93.33%) patients had subaortic ventricular septal defect (VSD), while three (6.6%) patients presented with doubly committed VSD. The most common type of right ventricular outflow tract (RVOT) obstruction was combined infundibular and valvular types, accounting for 34 cases (75.5%). Six patients had infundibular RVOT obstruction, while three patients (6.6%) had predominantly valvular pulmonary stenosis. We performed trans-right atrial repair in 33 patients. Right atrium-pulmonary artery approach was used in five patients (11.1%). The most common postoperative complication was right bundle branch block, seen in 14 patients, with a mortality rate of 2% in the early postoperative period. We achieved excellent early and midterm survival results and significant improvement in functions and disease-free quality of life. Conclusion: Intracardiac repair in adult TOF can be performed with low mortality, less residual RVOT obstruction, and need for revision of RVOT far less frequent by using the Jhajhria Infundibular Resection Adequacy Assessment technique (JIRAAT) to assess for adequacy of infundibular resection.

Development of non-viral vectors for neuronal-targeted delivery of CRISPR-Cas9 RNA-proteins as a therapeutic strategy for neurological disorders.

The aging population contributes to an increase in the prevalence of neurodegenerative diseases, such as Parkinson's disease (PD). Due to the progressive nature of these diseases and an incomplete understanding of their pathophysiology, current drugs are inefficient, with a limited efficacy and major side effects. In this study, CRISPR-Cas9 RNA-proteins (RNP) composed of a Cas9 nuclease and single-guide RNA were delivered with a non-viral targeted delivery system to rescue the PD-associated phenotype in neuronal cells. Here, we fused the cell-penetrating amphipathic peptide, PepFect14 (PF14), with a short fragment of the rabies virus glycoprotein (C2) previously shown to have an affinity towards nicotinic acetylcholine receptors expressed on neuronal cells and on the blood-brain barrier. The resultant peptide, C2-PF14, was used to complex with and deliver RNPs to neuronal cells. We observed that RNP/C2-PF14 complexes formed nanosized, monodispersed, and nontoxic nanoparticles that led to a specific delivery into neuronal cells. α-Synuclein (α-syn) plays a major role in the pathology of PD and is considered to be a target for therapy. We demonstrated that CRISPR/Cas9 RNP delivered by C2-PF14 achieved α-syn gene (SNCA) editing in neuronal cells as determined by T7EI assay and western blotting. Furthermore, RNP/C2-PF14 relieved PD-associated toxicity in neuronal cells in vitro. This is a proof-of-concept towards simple and safe targeted genome-editing for treating PD and other neurological disorders.

Outcome of surgical condition of neonates who underwent surgery: A prospective study from a tertiary care center.

Background: The neonatal period is a highly vulnerable time for an infant, who is completing many of the physiologic adjustments required for extra- uterine survival. If the neonate has a coexisting pathology which needs surgery, this challenge is magnified. Neonatal surgical conditions are unique in their type because some require early diagnosis, prompt surgery and postoperative care to improve the survival and outcome. Objective: The aim of this study was to know the clinical profile of congenital surgical conditions and to estimate the burden and outcome in special new born care unit. Method: The study population include 138 surgical neonate admitted in special new born care unit, department of paediatrics, Kamla Raja Hospital, Gajra Raja Medical College, Gwalior (M.P.) from April 2017 to April 2018 including six month follow up period. Results: Total admission in special new born care unit were 5378 out of which 138 (2.5%) neonates of surgical condition were admitted in the study period. Incidence of neonatal surgical condition was found to be 8.48%. Surgery was performed in 57 (41.30%) neonates. The Commonest neonatal surgical condition was constituted by gastrointestinal system (39.13%). Among gastrointestinal system anomalies, tracheoesophageal fistula were 28.6% of total gastrointestinal system cases. The most common surgical condition encountered was meningomyelocele, 23.36% of total cases. The survival of neonatal surgical condition in hospital was 52.89% and after six month follow up was 26.08%. The overall mortality with neonatal surgical condition in this study was 73.91%. Maternal age, antenatal care, history of congenital malformation, socioeconomic status, mode of delivery, prematurity, type of admission, single or multiple surgical condition, inotropic and ventilation support, post operative complication were significantly associated with final outcome of neonatal surgical condition. Conclusion: High mortality was found in neonates suffering from surgical conditions. Commonest anomaly includes conditions of gastrointestinal tract. Maternal age more than 35 year, poor antenatal care, prematurity, vaginal delivery, extra mural neonate, multiple surgical condition, inotropic and ventilation support and post operative complications were associated with increased mortality.

Intracardiac Repair in Late Adolescent and Adult Tetralogy of Fallot - Early and Midterm Results from a Tertiary Care Centre.

INTRODUCTION: In developing countries like India, it is common for late presentation of Tetralogy of Fallot (TOF) patients to a hospital as compared to that of developed countries. The objective of this study is to analyze the surgical outcome of TOF patients with age > 15 years. METHODS: This is a retrospective descriptive study of the surgical outcomes of 45 adult patients undergoing correction for TOF. Epidemiology, symptomology, and preoperative evaluation were performed. RESULTS: Most of the patients were male (33 [73%]). The median age was 21 years. A total of 42 (93.33%) patients had subaortic ventricular septal defect (VSD), while three (6.6%) patients presented with doubly committed VSD. The most common type of right ventricular outflow tract (RVOT) obstruction was combined infundibular and valvular types, accounting for 34 cases (75.5%). Six patients had infundibular RVOT obstruction, while three patients (6.6%) had predominantly valvular pulmonary stenosis. We performed trans-right atrial repair in 33 patients. Right atrium-pulmonary artery approach was used in five patients (11.1%). The most common postoperative complication was right bundle branch block, seen in 14 patients, with a mortality rate of 2% in the early postoperative period. We achieved excellent early and midterm survival results and significant improvement in functions and disease-free quality of life. CONCLUSION: Intracardiac repair in adult TOF can be performed with low mortality, less residual RVOT obstruction, and need for revision of RVOT far less frequent by using the Jhajhria Infundibular Resection Adequacy Assessment technique (JIRAAT) to assess for adequacy of infundibular resection.

Long-Term Clinicoradiological Outcomes of Cervical Fusion With Polyether Ether Ketone versus Cervical Disc Arthroplasty in a Double-Blinded Randomized Control Trial.

INTRODUCTION: In this double-blinded randomized controlled trial, we compare the long-term clinicoradiological effects of anterior cervical discectomy and fusion with polyether ether ketone implant (ACDF with PEEK) versus artificial cervical disc (cervical disc arthroplasty [CDA]) in patients with single and bilevel cervical degenerative disc disease (CDDD). AIMS: We aimed to compare the pain scores, cervical kinematics, and radiological outcomes in patients undergoing CDA and ACDF for single and bilevel CDDD over 5 years. SETTINGS AND DESIGN: This study was carried out from 2010 to 2019 in the Neurosurgery Department of a Tertiary Care Hospital attached to a Medical College in India. SUBJECTS AND METHODS: We enrolled 30 patients in each group. Clinical and radiological assessments were carried out for all patients over a period of 5 years. STATISTICAL ANALYSIS USED: Wilcoxon Signed-rank test, Mann-Whitney U-test, and Fischer's exact test were used for comparing the preoperative and follow-uP values. R software version 3.6.0 was used for statistical analysis. RESULTS: A significant improvement in the cervical range of motion, sagittal range of motion (ROM), and functional spinal unit (FSU) was observed in the CDA group at final follow-up, while the PEEK group reported a significant decrease in the ROM and FSU and the difference between the two groups was also found to be statistically significant. CONCLUSIONS: When assessed over 5 years following surgery, we found CDA to be superior with respect to ROM, FSU, overall cervical alignment, and maintenance of disc height when compared to ACDF with PEEK implant.

Surgical management and outcome of left ventricular pseudoaneurysm: our 11-year experience.

INTRODUCTION: Left ventricular (LV) pseudoaneurysm is an uncommon condition with a high risk of death due to spontaneous rupture. The symptoms are nonspecific and diagnosis is often delayed. Surgical repair is the treatment of choice despite associated operative mortality. AIM: Here we present a retrospective analysis of our experience in managing LV pseudoaneurysms over an 11-year period. MATERIAL AND METHODS: Between May 2009 and April 2020, 7 patients (6 males and 1 female) with LV pseudoaneurysm underwent surgical repair at our center. Hospital records were accessed to obtain relevant clinical information and treatment outcomes. The mean age was 41.86 years (range: 7-73 years). Etiologies were post-myocardial infarction (4 patients) and prior endocarditis/pericarditis (3 patients). Pseudoaneurysms were posterobasal in 4 patients and apical in 3 patients. All the patients underwent surgical repair with resection of pseudoaneurysm and patch repair of the ventricular wall defect. RESULTS: All patients tolerated surgery well with no perioperative mortality or morbidity. Clinical condition and echocardiographic findings remained stable in all patients over their follow-up period (3 months to 3 years). Mortality occurred in a 73-year-old patient with post-MI posterobasal pseudoaneurysm, 15 months after surgery due to acute exacerbation of chronic obstructive pulmonary disease. CONCLUSION: LV pseudoaneurysm is an entity that carries a high mortality risk. Timely diagnosis and early surgical intervention significantly improve the outcome.

Microplastics from lagooning sludge to composts as revealed by fluorescent staining- image analysis, Raman spectroscopy and pyrolysis-GC/MS.

Lagooning sludge (LS), which is used as soil amendment in Morocco, may contain microplastics (MPs). The aim of this study was to examine the effect of dewatering and co-composting of LS with green waste (GW) on the MPs' evolution. In this context the present study proposes fast-preliminary steps to detect plastics in lagooning sewage sludge before the extraction and identification process. We used pyrolysis GC/MS spectrometry to investigate the presence of chemical compounds possibly derived from plastics, and fluorescence staining by Nile Red to detect fluorescent particles suspected as plastics. Thereafter, we quantified the MPs particles after density fractionation and investigated their nature by Raman spectroscopy. RESULTS: indicated the presence of an average of 40.5 ± 11.9 × 103 MPs particles/kg (dry matter) and 36 ± 9.7 × 103 MPs particles/kg (dry matter) in fresh sludge and dewatered sludge respectively. Sludge dewatering in drying beds resulted a loss of small MPs (<500 µm). In co-composts, the quantity of MPs varied with the proportion of sewage sludge. The distribution of MPs types differentiated by colour and types (polypropylene, polyethylene, polyamide and polyester) evolved differently. Conventional co-composting did not have any effect on MPs quantity, indicating that they are not biodegradable under these temperature conditions, but it influenced their particle size. The risks of these pollutants after repeated field application and the possibility of their reduction through others co-composting procedures and techniques would be further investigated.

Correction of Niemann-Pick type C1 trafficking and activity with the histone deacetylase inhibitor valproic acid.

Niemann-Pick type C (NPC) disease is primarily caused by mutations in the NPC1 gene and is characterized by the accumulation of unesterified cholesterol and lipids in the late endosomal (LE) and lysosomal (Ly) compartments. The most prevalent disease-linked mutation is the I1061T variant of NPC1, which exhibits defective folding and trafficking from the endoplasmic reticulum to the LE/Ly compartments. We now show that the FDA-approved histone deacetylase inhibitor (HDACi) valproic acid (VPA) corrects the folding and trafficking defect associated with I1061T-NPC1 leading to restoration of cholesterol homeostasis, an effect that is largely driven by a reduction in HDAC7 expression. The VPA-mediated trafficking correction is in part associated with an increase in the acetylation of lysine residues in the cysteine-rich domain of NPC1. The HDACi-mediated correction is synergistically improved by combining it with the FDA-approved anti-malarial, chloroquine, a known lysosomotropic compound, which improved the stability of the LE/Ly-localized fraction of the I1061T variant. We posit that combining the activity of VPA, to modulate epigenetically the cellular acetylome, with chloroquine, to alter the lysosomal environment to favor stability of the trafficked I1061T variant protein can have a significant therapeutic benefit in patients carrying at least one copy of the I1061T variant of NPC1, the most common disease-associated mutation leading to NPC disease. Given its ability to cross the blood-brain barrier, we posit VPA provides a potential mechanism to improve the response to 2-hydroxypropyl-ß-cyclodextrin, by restoring a functional NPC1 to the cholesterol managing compartment as an adjunct therapy.

Molecular imaging reveals biodistribution of P-cadherin LP-DART bispecific and trafficking of adoptively transferred T cells in mouse xenograft model.

P-cadherin-LP-DART is a bispecific antibody targeting P-cadherin expressed on the tumor cells and CD3 on the T-cells. Previously we demonstrated the development and efficacy of P-cadherin-LP-DART in in vitro and in vivo models. Here, we evaluated the three pillars: exposure, targeting specificity and pharmacodynamic modulation for P-cadherin-LP-DART using fluorescence molecular tomography (FMT). Bispecific antibodies and T-cells were conjugated with a near-infrared fluorophores: VivoTag®680XL (VT680) and CellVue®NIR815 (CV815), respectively. In vitro binding and cytotoxic T-lymphocyte assay demonstrated that P-cadherin-LP-DART significantly retained its properties after VT680 conjugation. In vivo FMT imaging was performed to determine the bispecific biodistribution and T-cell trafficking in HCT-116 xenograft model. Peak tumor exposure (2.71%ID) was observed at 96 hr post-injection with measurable quantity even at 240 hr (1.46%ID) (Pillar 1). P-cadherin-LP-DART accumulation in tumor was 20-25 fold higher compared to Control-LP-DART demonstrating the targeting specificity (Pillar 2). Imaging after engraftment of CV815 labeled T-cells showed P-cadherin-LP-DART mediated T-cell trafficking in tumors (Pillar 3). This study harnessed the multichannel capability of FMT and demonstrated the targeting of drug and trafficking of T cells to tumors, simultaneously. Our results show the impact of molecular imaging in demonstrating three pillars of pharmacology, longitudinally and non-invasively.