Characterization of Juvenile Hormone Biosynthetic Enzymes in the Mosquito, Aedes aegypti

The juvenile hormones (JHs) are sesquiterpenoid compounds that play a central role in insect reproduction, development and behavior. They are synthesized and secreted by a pair of small endocrine glands, the corpora allata (CA), which are intimately connected to the brain. The enzymes involved in the biosynthesis of JH are attractive targets for the control of mosquito populations. This dissertation is a comprehensive functional study of five Aedes aegypti CA enzymes, HMG-CoA synthase (AaHMGS), mevalonate kinase (AaMK), phosphomevalonate kinase (AaPMK), farnesyl diphosphate synthase (AaFPPS) and farnesyl pyrophosphate phosphatase (AaFPPase). The enzyme AaHMGS catalyzes the condensation of acetoacetyl-CoA and acetyl-CoA to produce HMG-CoA. The enzyme does not require any co-factor, although its activity is enhanced by addition of Mg2+. The enzyme AaMK is a class I mevalonate kinase that catalyzes the ATP-dependent phosphorylation of mevalonic acid to form mevalonate 5-phosphate. Activity of AaMK is inhibited by isoprenoids. The enzyme AaPMK catalyzes the cation-dependent reversible reaction of phosphomevalonate and ATP to form diphosphate mevalonate and ADP. The enzyme AaFPPS catalyzes the condensation of isopentenyl diphosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) to form geranyl diphosphate (GPP) and farnesyl pyrophosphate (FPP). The enzyme AaFPPS shows an unusual product regulation mechanism, with chain length final product of 10 or 15 C depending on the metal cofactor present. The enzymes AaFPPase-1 and AaFPPase-2 efficiently hydrolyze FPP into farnesol, although RNAi experiments demonstrate that only AaFPPase-1 is involved in the catalysis of FPP into FOL in the CA of A. aegypti. This dissertation also explored the inhibition of the activity of some of the JH biosynthesis enzymes as tools for insect control. We described the effect of N-acetyl-S-geranylgeranyl-L-cysteine as a potent inhibitor of AaFPPase 1 and AaFPPase-2. In addition, inhibitors of AaMK and AaHMGS were also investigated using purified recombinant proteins. The present study provides an important contribution to the characterization of recombinant proteins, the analysis of enzyme kinetics and inhibition constants, as well as the understanding of the importance of these five enzymes in the control of JH biosynthesis rates.

Tratamento de cascalho de perfuração utilizando sistemas microemulsionados

The oil industry is one of the activities that generates more waste to the environment. The drill cuttings is a waste generated in large quantities in the drilling process and that may cause environmental damage such as soil contamination and consequently the contamination of groundwater if disposed of without prior treatment. Arises the need to develop scientific activities and research ways to adapt these wastes the current environmental standards. In the case of solid wastes, the NBR 10004: 2004 of the Brazilian Association of Technical Standards (ABNT) classifies them into class I waste (hazardous) and class II (not dangerous), which determines which wastes may or may not be discarded in the environment without causing environmental impact. This study presents a novel alternative for treating drill cuttings, where this waste was classified as class I (Abreu & Souza, 2005), mainly by removing the n-paraffin present in it, since this arises when using drilling fluids base oil. Using microemulsion systems promotes the removal of this contaminant drill cuttings samples from wells located in Alto do Rodrigues - RN. Initially, we determined the concentration of paraffin using infrared method in samples were extracted with ultrasound, we obtained a paraffin concentration in the range from 36.59 to 43.52 g of paraffin per kilogram of cuttings. Used two microemulsion systems containing two nonionic surfactants from different classes, one is an alcohol ethoxylated (UNTL-90) and the other an nonylphenol ethoxylated (RNX 110). The results indicated that the system UNTL-90 surfactant has better efficiency than the system with RNX 110. The study of the influence of contact time at the extraction showed that for times greater than 25 minutes has a tendency to increase the percentage extraction with increasing contact time. It was also observed that the extraction is fast because at 1 minute contact has 22.7% extraction. The reuse of the microemulsion system without removing the paraffin extracted in previous steps, showed reduction of 29.32 in percentage of extraction by comparing the first and third extraction, but by comparing the first and second extractions reduction is 8.5 in percentage extraction, so the systems reuse optimization can be an option for economically viable removing paraffin from cuttings. The extraction with shaking is more effective in the treatment of cuttings, reaching the extraction percentage of 87.04%, that is, obtaining a drill cuttings with 0.551% paraffin. Using the percentage of paraffin employed in non-aqueous drilling fluids and fluid maximum limit on cuttings for disposal established by the Environmental Protection Agency of the United States (US EPA), one arrives at the conclusion that the level of paraffin on gravel cannot exceed 3.93%. Conclude that the amount of paraffin in the treated cuttings with the microemulsion system with shaking is below the established by US EPA, showing that the system used was efficient in removing the paraffin from the drill cuttings.

Ethics in the provision of removable partial dentures

Aim: To investigate the construction of cobalt-chromium removable partial dentures by commercial private dental laboratories. Methods: Ninety master casts for fabrication of cobalt-chromium removable partial dentures were obtained from three commercial laboratories randomly selected. Casts were assessed for dental arch treated, Kennedy classification, cast surveying, denture design information provided by the dentist, and mouth preparation (rest seat, guiding plane and retentive area). Dental technicians answered a questionnaire regarding qualification of assisted dentists, monthly number of framework castings, and use of dental surveyor. Mouth preparation was compared among laboratories using Kruskal-Wallis test (α=0.05). Results: The percentage of Kennedy class I was 16%, class II 19%, class III 56%, and class IV 9%. The majority of master cats (51%) examined was sent to dental laboratories without any design information and did not comply with ethical guidelines in the provision of RPD. Approximately half of the casts were considered “inappropriate” for guiding planes and retentive areas. One of the laboratories presented all casts “inappropriate” for rest seat distribution (p<0.001). Conclusions: Mouth preparation frequently failed for guiding planes, retentive areas and distribution of rest seats. It is necessary to provide students with adequate clinical experience at the dental school environment, which will actually be carried into the practice of dentistry.

Ethics in the provision of removable partial dentures

Aim: To investigate the construction of cobalt-chromium removable partial dentures by commercial private dental laboratories. Methods: Ninety master casts for fabrication of cobalt-chromium removable partial dentures were obtained from three commercial laboratories randomly selected. Casts were assessed for dental arch treated, Kennedy classification, cast surveying, denture design information provided by the dentist, and mouth preparation (rest seat, guiding plane and retentive area). Dental technicians answered a questionnaire regarding qualification of assisted dentists, monthly number of framework castings, and use of dental surveyor. Mouth preparation was compared among laboratories using Kruskal-Wallis test (α=0.05). Results: The percentage of Kennedy class I was 16%, class II 19%, class III 56%, and class IV 9%. The majority of master cats (51%) examined was sent to dental laboratories without any design information and did not comply with ethical guidelines in the provision of RPD. Approximately half of the casts were considered “inappropriate” for guiding planes and retentive areas. One of the laboratories presented all casts “inappropriate” for rest seat distribution (p<0.001). Conclusions: Mouth preparation frequently failed for guiding planes, retentive areas and distribution of rest seats. It is necessary to provide students with adequate clinical experience at the dental school environment, which will actually be carried into the practice of dentistry.

Inter-relação entre lesão cervical não cariosa e comprometimento periodontal: diagnóstico, tratamento e previsibilidade - estudo clínico prospectivo e laboratorial

The coexistence of gingival recession (GR) with root coverage indication and non-carious cervical lesions (LCNC) generates the need for a protocol that respects and promotes health of dental and periodontal tissues and allows treatment predictability. The main objectives of this theses were: (1) verify, through clinical evaluations, the connective tissue graft for root coverage on direct and indirect restorations made of ceramic resin; (2) analyze the influence of the battery level of the LED curing unit in the composite resin characteristics; (3) assess the influence of restorative materials, composite resin and ceramics, on the viability of gingival fibroblasts from primary culture. Nine patients with good oral hygiene and occlusal stability diagnosed with LCNCs the anterior teeth including premolars associated with gingival recession (class I and II of Miller) and only gingival recession were selected. After initial clinical examination, occlusal adjustment was performed and the patients had their teeth randomized allocated on direct composite resin restoration of LCNC, polishing and GR treatment with connective tissue graft and advanced coronally flap CR group (n = 15); and indirect ceramic restoration of the LCNC's and GR treatment (CTG+CAF) Group C (n = 15). The GR presented teeth with no clinically formed LCNCs cavity were treated using (CTG+CAF) being the control group (n = 15). Sorption and solubility tests, analysis of the degree of conversion and diametral tensile strength were performed in composite resin samples (n = 10) photoactivated by 100, 50 and 10% battery charge LED unit. The viability of fibroblasts on composite resin, ceramics and dentin disks (n = 3) was examined. Clinical follow-up was performed for three months. The data obtained at different stages were tabulated and subjected to analysis for detection of normal distribution and homogeneity. The results showed that: the LED unit with 10% battery affects the characteristics of the composite resin; restorative materials present biocompatibility with gingival fibroblasts; and the association of surgical and restorative treatment of teeth affected by NCCL and GR presents successful results at 3-month follow-up.

Estudio en aves silvestres de canto ("Passeriformes") de las moléculas MHC y reclasificación molecular de la familia "Carduelini"

The Major Histocompatibility Complex (MHC) comprises the most polymorphic loci in animals. MHC plays an important role during the first steps of the immune response in vertebrates. In humans, MHC molecules (also named human leukocyte antigens, HLA) were initially regarded as class I or class II molecules. Each of them, presents to different T cells subsets. MHC class I molecules, are heterodimers in which the heavy chain (alpha) has three extracellular domains, two of which (alpha 1 and alpha 2) are polymorphic and conform the antigen recognition sites (ARS). The ARS is thought to be subjected to balancing selection for variability, which is the cause of the very high polymorphism of the MHC molecules. Different pathogenic epitopes would be the evolutionary force causing balancing selection. MHC class I genes have been completely sequenced (α1 and α2 protein domains) and thoroughly studied in Gallus gallus (chicken) as well as in mammals. In fact, the MHC locus was first defined in chicken, specifically in the highly consanguineous variety „Leghorn‟. It has been found that, in the case of chickens the MHC genetic region is considerably smaller than it is in mammals (remarkably shorter introns were found in chickens), and is organized quite differently. The noteworthy presence of short introns in chickens; supported the hypothesis that chicken‟s MHC represented a „minimal essential MHC‟. Until now, it has been assumed that chicken (order Galliformes) MHC was similar to all species included in the whole class Aves...

Difficult Labor: Troublesome Company at the New Birth of Freedom

Extended contribution to a roundtable on Mark A. Lause's <i>Free Labor: The Civil War and the Making of an American Working Classi>, emphasizing the wartime labor movement's great difficulty in responding to rapid industrialization brought on by the war, and to the increasing diversity of the labor force brought about by mass immigration.

Mechanismen kleiner regulatorischer RNAs in Dictyostelium discoideum: Charakterisierung und Anwendung

Die RNA-Interferenz (RNAi) ist ein in Eukaryoten weit verbreiteter Mechanismus, der die transkriptionelle oder posttranskriptionelle Stilllegung von Genen beschreibt. Die Spezifität wird dabei durch die Sequenz einer kleinen, nicht-kodierenden RNA gewährleistet. Diese RNA leitet einen Effektorkomplex, dessen Zentrum immer von einem Argonautenprotein gebildet wird, üblicherweise zu einer komplementären mRNA. In der Folge kommt es zum Abbau des Transkripts oder zur Inhibierung der Translation. Aktuelle Veröffentlichungen konnten zudem das Aktivitätsprofil der Argonautenproteine beträchtlich erweitern: Im Zellkern vorkommende Argonautenproteine wurden beispielsweise mit Spleißvorgängen, der Promotorkontrolle von Genen und der DNA-Reparatur in Verbindung gebracht. In den letzten Jahren konnten weitreichende Kenntnisse bezüglich der Kontrolle einiger transposabler Elemente durch RNAi sowie der Biogenese kleiner regulatorischer RNAs in Dictyostelium discoideum und anderen Organismen gewonnen werden. Ein Fokus dieser Arbeit lag zunächst auf der Charakterisierung des Argonautenproteins AgnB und der Identifikation von Interaktionspartnern. Es konnte gezeigt werden, dass AgnB zumindest partiell im Zellkern der Amöbe lokalisiert und dort vermutlich regulatorische Aufgaben wahrnimmt. Gestützt wurde diese Annahme durch die massenspektrometrische und Western Blot basierte Detektion nukleärer Bindungspartner. Weiterführende Analysen konnten AgnB zudem als positiven Regulator für drei entwicklungsregulierte Gene beschreiben und so die Verbindung zum Prozess der RNA activation in der Amöbe herstellen. Identifizierte posttranslationale Modifikationen könnten diesbezüglich die Aktivität des Argonauten steuern. Mit Hilfe von Crosslink-RNA-Immunopräzipitation und anschließender Hochdurchsatz-sequenzierung oder der Northern Blot basierten Auswertung konnte eine Assoziation von AgnB und der Class I RNAs gezeigt werden. Diese Klasse umfasst nicht-kodierende RNAs mit einer Länge von etwa 42 bis 65 Nukleotiden und wurde bisher nicht als Substrat für die RNAi-Maschinerie in D. discoideum angesehen. Ein weiterer Teil dieser Arbeit beschäftigte sich mit dem Einfluss von AgnA und AgnB auf die Promotorbereiche des D. discoideum Retrotransposons DIRS-1. Im Verlauf der Untersuchungen konnte beobachtet werden, dass die Anordnung entgegengesetzt operierender DIRS-1 Promotor-sequenzen für die Stilllegung eines Reportergens ausreichend war. Darauf aufbauend konnte ein DIRS-1 basiertes knockdown System etabliert werden. Mit Hilfe dieses Systems konnten die Proteinmengen ausgewählter Zielgene so effektiv reduziert werden, dass die entsprechenden Stämme den Phänotyp des korrespondierenden knockout Stammes zeigten. Darüber hinaus war es möglich die Proteinreduktion zu modulieren, um so beispielsweise dosisabhängige Effekte zu registrieren. Vorangegangene Arbeiten zur Biogenese von micro (mi)RNAs konnten das RNA-bindende Protein RbdB als eine Hauptkomponente für die Prozessierung maturer miRNAs identifizieren. Der miRNA defiziente RbdB- Stamm wurde in dieser Arbeit zur Identifikation putativer miRNA Ziele genutzt. Dazu wurde sowohl das Transkriptom des D. discoideum Wildtyps als auch des rbdB knockout Stammes in hohem Durchsatz sequenziert, um so differentiell exprimierte Gene zu detektieren. Vielversprechende Kandidaten wurden mittels qRT-PCR verifiziert. Dabei wurde unter anderem ein putativer Transkriptionsfaktor als miRNA target identifiziert, der eine Vielzahl weiterer Gene regulieren könnte. Abschließend konnte in dieser Arbeit gezeigt werden, dass RbdB ebenfalls für die Generierung von small interfering (si)RNAs aus strukturierten Loci von Bedeutung ist. Dies weist auf mindestens zwei unterschiedliche Mechanismen zur siRNA Prozessierung in D. discoideum hin.

Mitral Valve Surgery for Rheumatic Lesions in Young Patients

BACKGROUND: The appropriateness of rheumatic mitral valve repair remains controversial due to the risks of recurrent mitral dysfunction and need for reoperation. The aims of this study were to determine the overall short- and long-term outcomes of pediatric rheumatic mitral valve surgery in our center. METHODS: Single-center, observational, retrospective study that analyzed the results of rheumatic mitral valve surgery in young patients, consecutively operated by the same team, between 1999 and 2014. RESULTS: We included 116 patients (mean age = 12.6 ± 3.5 years), of which 66 (57%) were females. A total of 116 primary surgical interventions and 22 reoperations were performed. Primary valve repair was possible in 86 (74%) patients and valve replacement occurred in 30 (26%). Sixty percent of the patients were followed up beyond three months after surgery (median follow-up time = 9.2 months [minimum = 10 days; maximum = 15 years]). Long-term clinical outcomes were favorable, with most patients in New York Heart Association functional class I (89.6%) and in sinus rhythm (85%). Freedom from reoperation for primary valve repair at six months, five years, and ten years was 96.4% ± 0.25%, 72% ± 0.72%, and 44.7% ± 1.34%, respectively. Freedom from reoperation for primary valve replacement at six months, five years, and ten years was 100%, 91.7% ± 0.86%, and 91.7% ± 0.86%, respectively. Mitral stenosis as the primary lesion dictated early reintervention. CONCLUSIONS: Despite the greater rate of reoperation, especially when the primary lesion was mitral stenosis, rheumatic mitral valve repair provides similar clinical outcomes as compared with replacement, with the advantage of avoiding anticoagulation.

Highway 60 Reconstruction Project : Final Environmental Impact Statement, 2004

The proposed reconstruction of Trunk Highway 60 is considered a Federal Class I Action because of the potential for significant impacts on the natural and physical environment. An Environmental Impact Statement (EIS) is a full disclosure document that discusses the environmental impacts of a proposed Class I action. The Draft EIS, which was distributed in July 2002, is incorporated by reference herein and made a part of the Final EIS.

Development of Medical Devices

The medical devices sector helps save lives by providing innovative health care solutions regarding diagnosis, prevention, monitoring, treatment, and alleviation. Medical devices are classified into 1 of 3 categories in the order of increasing risk: Class I, Class II, and Class III.1 Medical devices are distinguished from drugs for regulatory purposes based on mechanism of action. Unlike drugs, medical devices operate via physical or mechanical means and are not dependent on metabolism to accomplish their primary intended effect.

Continuous acquisition of MHC:peptide complexes by recipient cells contributes to the generation of anti-graft CD8+T cell immunity

Understanding the evolution of the direct and indirect pathways of allorecognition following tissue transplantation is essential in the design of tolerance-promoting protocols. On the basis that donor bone marrow-derived antigen presenting cells are eliminated within days of transplantation, it has been argued that the indirect response represents the major threat to long term transplant survival, and is consequently the key target for regulation. However, the detection of MHC transfer between cells, and particularly the capture of MHC:peptide complexes by dendritic cells, led us to propose a third, semi-direct, pathway of MHC allorecognition. Persistence of this pathway would lead to sustained activation of direct pathway T cells, arguably persisting for the life of the transplant. In this study, we focused on the contribution of acquired MHC class I, on recipient DCs, during the life span of a skin graft. We observed that MHC class I acquisition by recipient DCs occurs for at least one month following transplantation and may be the main source of alloantigen that drives CD8+ cytotoxic T cell responses. In addition, acquired MHC class I-peptide complexes stimulate T cell responses in vivo further emphasizing the need to regulate both pathways to induce indefinite survival of the graft.

NK Cells of Kidney Transplant Recipients Display an Activated Phenotype that Is Influenced by Immunosuppression and Pathological Staging

To explore phenotype and function of NK cells in kidney transplant recipients, we investigated the peripheral NK cell repertoire, capacity to respond to various stimuli and impact of immunosuppressive drugs on NK cell activity in kidney transplant recipients. CD56(dim) NK cells of kidney transplanted patients displayed an activated phenotype characterized by significantly decreased surface expression of CD16 (p=0.0003), CD226 (p<0.0001), CD161 (p=0.0139) and simultaneously increased expression of activation markers like HLA-DR (p=0.0011) and CD25 (p=0.0015). Upon in vitro stimulation via Ca++-dependent signals, down-modulation of CD16 was associated with induction of interferon (IFN)-gamma expression. CD16 modulation and secretion of NFAT-dependent cytokines such as IFN-gamma, TNF-alpha, IL-10 and IL-31 were significantly suppressed by treatment of isolated NK cells with calcineurin inhibitors but not with mTOR inhibitors. In kidney transplant recipients, IFN-gamma production was retained in response to HLA class I-negative target cells and to non-specific stimuli, respectively. However, secretion of other cytokines like IL-13, IL-17, IL-22 and IL-31 was significantly reduced compared to healthy donors. In contrast to suppression of cytokine expression at the transcriptional level, cytotoxin release, i.e. perforin, granzyme A/B, was not affected by immunosuppression in vitro and in vivo in patients as well as in healthy donors. Thus, immunosuppressive treatment affects NK cell function at the level of NFAT-dependent gene expression whereby calcineurin inhibitors primarily impair cytokine secretion while mTOR inhibitors have only marginal effects. Taken together, NK cells may serve as indicators for immunosuppression and may facilitate a personalized adjustment of immunosuppressive medication in kidney transplant recipients.

Doença enxerto-versus hospedeiro: determinação da expressão de genes e alterações de vias associadas ao sistema imunológico usando metodos da biofísica

O transplante de medula óssea (TMO) é um procedimento terapêutico importante em casos relacionados à pacientes com leucemia ou linfoma. Em decorrência desse processo, uma reação conhecida como doença enxerto-versus-hospedeiro (GVHD) pode ocorrer em pacientes susceptíveis como conseqüência da presença de células imunocompetentes do doador. Entretanto, não existe um modelo para descrever completamente as ações relacionadas ao mecanismo imunológico da GVHD desde a fase que inicializa a doença até a fase efetora. O Objetivo geral deste estudo é a investigação da resposta imunológica considerando-se o sistema HLA (antígenos leucocitários humano) em pacientes que desenvolveram a GVHD em decorrência do TMO. O National Cancer Institute (NCI) – Pathway interaction Database e Reactome foram usados como bases de dados com o objetivo de se estudar a expressão de genes e vias relacionados às Classes I e II do sistema HLA (antígenos leucocitários humano). O estudo considerou a mudança de expressão de genes relacionados às 17 vias do sistema imunológico com potencialidade para se expressar em pacientes que desenvolveram a GVHD associada à TMO. Dados referentes aos transcriptomas foram obtidos utilizando-se a plataforma GPL570 Affymetrix Genoma Humano U133 Plus. A atividade relativa foi usada para determinar as alterações das vias em amostras de GVHD em relação ao controle. As análises foram realizadas utilizando-se o software Via Complex e Bioconductor. Observou-se aumento significativo da expressão de genes ralacionados às vias do sistema imune adaptativo, antígenos associados às Classe I e II do HLA, fosforilação de CD3 e CD247, sinalização dos receptores de células T em CD4+ nativas e ativação de NF-kapa β nas células B. Também observou-se alterações significativas na mudança de expressão dos genes associados às vias relacionadas à super família de moléculas B7:CD28\CTLA-4 quando comparadas ao controle. Isso pode indicar a necessidade de geração de um segundo sinal co-estimulador em GVHD, acionado pelas moléculas dessa super família. O aumento da expressão do gene CD69 nas amostras experimentais caracteriza a ativação celular e, portanto, a sinalização de estímulos em GVHD. Os achados obtidos neste estudo contribuem para melhor elucidar o mecanismo imunopatogênico associado à GVHD. P

Scheduling of track inspection and maintenance activities in railroad networks

The U.S. railroad companies spend billions of dollars every year on railroad track maintenance in order to ensure safety and operational efficiency of their railroad networks. Besides maintenance costs, other costs such as train accident costs, train and shipment delay costs and rolling stock maintenance costs are also closely related to track maintenance activities. Optimizing the track maintenance process on the extensive railroad networks is a very complex problem with major cost implications. Currently, the decision making process for track maintenance planning is largely manual and primarily relies on the knowledge and judgment of experts. There is considerable potential to improve the process by using operations research techniques to develop solutions to the optimization problems on track maintenance. In this dissertation study, we propose a range of mathematical models and solution algorithms for three network-level scheduling problems on track maintenance: track inspection scheduling problem (TISP), production team scheduling problem (PTSP) and job-to-project clustering problem (JTPCP). TISP involves a set of inspection teams which travel over the railroad network to identify track defects. It is a large-scale routing and scheduling problem where thousands of tasks are to be scheduled subject to many difficult side constraints such as periodicity constraints and discrete working time constraints. A vehicle routing problem formulation was proposed for TISP, and a customized heuristic algorithm was developed to solve the model. The algorithm iteratively applies a constructive heuristic and a local search algorithm in an incremental scheduling horizon framework. The proposed model and algorithm have been adopted by a Class I railroad in its decision making process. Real-world case studies show the proposed approach outperforms the manual approach in short-term scheduling and can be used to conduct long-term what-if analyses to yield managerial insights. PTSP schedules capital track maintenance projects, which are the largest track maintenance activities and account for the majority of railroad capital spending. A time-space network model was proposed to formulate PTSP. More than ten types of side constraints were considered in the model, including very complex constraints such as mutual exclusion constraints and consecution constraints. A multiple neighborhood search algorithm, including a decomposition and restriction search and a block-interchange search, was developed to solve the model. Various performance enhancement techniques, such as data reduction, augmented cost function and subproblem prioritization, were developed to improve the algorithm. The proposed approach has been adopted by a Class I railroad for two years. Our numerical results show the model solutions are able to satisfy all hard constraints and most soft constraints. Compared with the existing manual procedure, the proposed approach is able to bring significant cost savings and operational efficiency improvement. JTPCP is an intermediate problem between TISP and PTSP. It focuses on clustering thousands of capital track maintenance jobs (based on the defects identified in track inspection) into projects so that the projects can be scheduled in PTSP. A vehicle routing problem based model and a multiple-step heuristic algorithm were developed to solve this problem. Various side constraints such as mutual exclusion constraints and rounding constraints were considered. The proposed approach has been applied in practice and has shown good performance in both solution quality and efficiency.