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As an important model animal, fruit fly is characterized by outstanding genetic characteristics, relatively perfect nervous system, rapid reproduction, and low cost. Thus, it has been applied in the research on neuropsychiatric disorders in recent years, showing great potential in life science. The incidence of neuropsychiatric disorders has been on the rise, and the disorders have high disability rate and low case fatality rate. The global drug demand for such diseases is second only to cardiovascular and cerebrovascular diseases. At the moment, the demand of the drugs for the diseases have been rising, and it is an urgent task to develop related drugs. However, the research and development of the drugs are time-intensive and have a high failure rate. A suitable animal model can help shorten the time for drug screening and development, thereby reducing the cost and failure rate. This study reviews the application of fruit flies in several common neuropsychiatric disorders, which is expected to provide new ideas for the research and application of the model animals in traditional Chinese medicine.
Subject(s)
Animals , Medicine, Chinese Traditional , Drugs, Chinese Herbal/therapeutic use , Models, Animal , Cerebrovascular DisordersABSTRACT
RESUMEN Introducción: Desde 1980 se conocen casos de síndromes neuropsiquiátricos infantiles en el mundo y su concepto ha evolucionado con cambios en las definiciones de 1995 (PITANDS: trastornos neuropsiquiátricos pediátricos autoinmunes precipitados por infección), 1998 (PANDAS: síndrome neuropsiquiátrico autoinmune pediátrico asociado con la infección por estreptococos), 2010 (PANS: síndrome pediátrico neuropsiquiátrico de inicio agudo) y 2012 (CANS: síndromes neuropsiquiátricos agudos de los niños). A pesar de que se conoce desde hace más de 20 años, aún es una enfermedad que suele pasar inadvertida para muchos profesionales de la salud. Objetivo: Sensibilizar a la comunidad médica acerca de la identificación de la enfermedad y disminuir la morbilidad asociada con un diagnóstico tardío. Caso clínico: Una niña de 6 años consultó a urgencias por trastorno de rechazo alimentario. En el tratamiento hospitalario se identificó historia clínica con criterios diagnósticos de PANS-PANDAS. Mostraba un curso clínico recurrente-remitente, tal y como describe la literatura, con pobre respuesta a los tratamientos de primera línea. Conclusiones: En todo niño en edad escolar que se presente con trastorno obsesivo compulsivo o trastornos alimentarios, con sin otros síntomas, se debe evaluar y descartar su asociación con PANS-CANS.
ABSTRACT Introduction: Since 1980, there have been known cases of childhood neuropsychiatric syndromes in the world and its concept has evolved with changes in the definitions in 1995 (PITANDs - paediatric infection-triggered autoimmune neuropsychiatric disorders), 1998 (PANDAS - paediatric autoimmune neuropsychiatric syndrome associated with streptococci infection), 2010 (PANS - paediatric acute-onset neuropsychiatric syndrome) and 2012 (CANS - childhood acute neuropsychiatric syndrome). Despite being known for more than 20 years, it is still an illness that often goes unnoticed by many health professionals. Objective: To sensitise the medical community about the identification of the disease and reduce the morbidity associated with a late diagnosis. Clinical case: A 6-year-old schoolgirl brought to the emergency department due to her refusal to eat. In the hospital treatment, a clinical history was identified with PANS-PANDAS diagnostic criteria. She exhibited a relapsing-remitting clinical course, as described in the literature, with poor response to first-line treatments. Conclusions: In all school-age child presenting with obsessive compulsive disorder or eating disorders, with other symptoms or not, a possible link to PANS-CANS should be evaluated and ruled out.
ABSTRACT
El objetivo del estudio fue investigar las funciones ejecutivas (fe) y los trastornos neuropsiquiátricos (tn) en pacientes con enfermedad de Alzheimer (ea), en función del grado de severidad, en comparación con sujetos sin deterioro cognitivo (ssdc). Se estudia-ron 50 pacientes con ea y 60 ssdc en un estudio no experimental-transversal con un muestreo no probabilístico basado en una serie de criterios de inclusión. Se aplicó la batería de evaluación frontal y el inventario neuropsiquiátrico, y los resultados mostraron que existe una relación negativa de intensidad moderada entre las fe y lostn, con independencia del grado de severidad de la ea. Se puede concluir que, en fase leve, la euforia se relaciona tanto con la sensibilidad hacia la interferencia como con el control inhibitorio; en fase moderada, la irritabilidad se relaciona de manera positiva con la sensibilidad hacia la interferencia, y en la fase moderadamente grave, la programación motora se relaciona de manera negativa con la agitación.
The aim of the study was to investigate executive func-tions (ef) and neuropsychiatric disorders (nd) in pa-tients with Alzheimer's disease (ad) according to the degree of severity, compared to subjects without cognitive impairment (swci). Fifty patients with ad and 60 swci were studied in a non-experimental-cross-sectional study with non-probabilistic sampling based on a series of inclusion criteria. The frontal evaluation battery and the neuropsychiatric inventory were applied and the results showed that there is a negative relationship of moderate intensity between ef and nd, regardless of the degree of severity of ad. It can be concluded that, in the mild phase, euphoria is related both to sensitivity towards interference and to inhibitory control; in moderate phase irritability is positively related to sensitivity towards interference; and in the moderately severe phase, motor programming is nega-tively related to agitation.
O objetivo do estudo foi investigar as funções executivas (fe) e os transtornos neuropsiquiátricos (tn) em pacien-tes com doença de Alzheimer (da) de acordo com o estágio de gravidade, em comparação com sujeitos sem comprometimento cognitivo (sscc). Foram avaliados 50 pacientes com da e 60 sscc em um estudo transversal não experimental com amostragem não probabilística baseada em uma série de critérios de inclusão. A bateria de avaliação frontal e o inventário neuropsiquiátrico foram aplicados e os resultados mostraram que existe uma relação negativa de intensidade moderada entre a fe e os tn, independente do estágio de gravidade da da. Pode-se concluir que, na fase leve, a euforia está rela-cionada tanto à sensibilidade à interferência quanto ao controle inibitório; na fase moderada, a irritabilidade está positivamente relacionada à sensibilidade à inter-ferência e; na fase moderadamente grave, a programação motora está negativamente relacionada à agitação.
Subject(s)
Humans , Patients , Training Support , Disease , Sensitivity and Specificity , Alzheimer Disease , Cognitive DysfunctionABSTRACT
The microbiota-gut-brain axis is a bidirectional signaling mechanism between the gastrointestinal tract and the central nervous system. The complexity of the intestinal ecosystem is extraordinary; it comprises more than 100 trillion microbial cells that inhabit the small and large intestine, and this interaction between microbiota and intestinal epithelium can cause physiological changes in the brain and influence mood and behavior. Currently, there has been an emphasis on how such interactions affect mental health. Evidence indicates that intestinal microbiota are involved in neurological and psychiatric disorders. This review covers evidence for the influence of gut microbiota on the brain and behavior in Alzheimer disease, dementia, anxiety, autism spectrum disorder, bipolar disorder, major depressive disorder, Parkinson's disease, and schizophrenia. The primary focus is on the pathways involved in intestinal metabolites of microbial origin, including short-chain fatty acids, tryptophan metabolites, and bacterial components that can activate the host's immune system. We also list clinical evidence regarding prebiotics, probiotics, and fecal microbiota transplantation as adjuvant therapies for neuropsychiatric disorders.
Subject(s)
Humans , Depressive Disorder, Major , Autism Spectrum Disorder , Gastrointestinal Microbiome , Brain , EcosystemABSTRACT
Objective Metformin ( MET) is the first-line medications of choice for patients with type 2 diabetes. In recent years,a large number of studies at home and abroad have shown that metformin not only has significant effects in controlling blood sugar level of diabetic patients and intervening in impaired glucose tolerance,but also has ＂ magical＂ effects in in the field of hypoglycemic outside. In addition to the early discovery that metformin can be used to treat metabolic syndrome and polycystic ovary syndrome,control body weight,blood pressure,blood lipid,and reduce the risk of cardiovascular events,metformin can also play anti-tumor,anti-inflammatory effects,delay aging,and treat mental disorders such as depression. This article reviews the effect of metformin in the field out of hypoglycemic.
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Investigating the pathophysiological mechanisms underlying brain disorders is a priority if novel therapeutic strategies are to be developed. In vivo studies of animal models and in vitro studies of cell lines/primary cell cultures may provide useful tools to study certain aspects of brain disorders. However, discrepancies among these studies or unsuccessful translation from animal/cell studies to human/clinical studies often occur, because these models generally represent only some symptoms of a neuropsychiatric disorder rather than the complete disorder. Human brain slice cultures from postmortem tissue or resected tissue from operations have shown that, in vitro, neurons and glia can stay alive for long periods of time, while their morphological and physiological characteristics, and their ability to respond to experimental manipulations are maintained. Human brain slices can thus provide a close representation of neuronal networks in vivo, be a valuable tool for investigation of the basis of neuropsychiatric disorders, and provide a platform for the evaluation of novel pharmacological treatments of human brain diseases. A brain bank needs to provide the necessary infrastructure to bring together donors, hospitals, and researchers who want to investigate human brain slices in cultures of clinically and neuropathologically well-documented material.
Subject(s)
Humans , Brain , Brain Diseases , Drug Therapy , Tissue Culture TechniquesABSTRACT
Despite some innate limitations, animal models are a potent investigative tool when used to model specific symptoms of a disorder. For example, MK-801, an N-methyl-D-aspartate receptor antagonist, is used as a pharmacological tool to induce symptoms found in some neuropsychiatric disorders. However, a close examination of literature suggests that the application window of MK-801 doses is relatively narrow between individual behavioral paradigms, necessitating careful characterization of the evoked behavioral aberrations and the doses used to induce them. Moreover, variation in behaviors depending on the animal strain, gender of the subject, and the timing of administration is observed, making it difficult to compare the behavioral characteristics reported in different studies. We aim to characterize the behavioral aberrations induced by different doses of MK-801 in CD-1 mice and create a ready reference for future studies. We used CD-1 mice to recapitulate behavioral impairments resulting from acute administration of MK-801. In 0.1 mg kg⁻¹, we observed diminished spontaneous alteration during the Y-maze test, while 0.12 mg kg⁻¹ resulted in hyperlocomotion and social deficit. Mice treated with 0.2 and 0.3 mg kg⁻¹ of MK-801 demonstrated a decreased self-grooming. Finally, all doses significantly impaired cliff avoidance behaviors suggesting increased impulsivity. These results affirm that MK-801 can effectively model various symptoms of different neuropsychiatric disorders in a dose-dependent manner. The observed sensitivity against spatial-memory impairment and impulsive behaviors at low concentration of MK-801 suggest that MK801 may modulate cognitive function and impulsivity in even lower concentration before it can modulate other behavioral domains.
Subject(s)
Animals , Mice , Avoidance Learning , Cognition , Dizocilpine Maleate , Impulsive Behavior , Models, Animal , N-MethylaspartateABSTRACT
Drug addiction has not only seriously damaged the addicts' health, but also caused a huge social and economic burden, threatening public safety. Meanwhile, the number of drug addicts continues to rise and the relapse rate remains high. Actually drug addiction is a kind of chronic brain disease. It is of no avail to reduce the psychological requirement of addicts via alternative drug treatments such as methadone, non-pharmacological treatments, surgical treatment, mindfulness therapy, and repeated transcranial magnetic stimulation etc. In fact, the rate of relapse is still high, accompanied by a series of huge limitations. Therefore, it is of great importance to find new therapeutic drugs and methods for drug addiction. In recent years, numerous studies of intestinal flora and neuropsychiatric diseases have been reported, gradually revealing the link between the intestinal microbiota and the nervous system. With the discovery of the "microbiota-gut-brain axis", the effect of gut microbiota on related aspects of drug addiction has won increasing attention. As a result, this thesis will review the research progress of the relationship between gut microbiota, neuropsychiatric diseases and drug addiction by means of looking up domestic and foreign documents.
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Background: Neuropsychiatric disorders are quite prevalent in the community and are on the rise due to changing lifestyles and lack of social support. Most of the published data is from hospital, which calls for more community-based studies to measure the actual burden of this menace. Aim & Objective: To estimate the prevalence of neuropsychiatric disorders in district Dehradun and to find out association of socio-demographic correlates with neuropsychiatric disorders Settings and Design: This community based cross-sectional survey was conducted in selected rural and urban areas of district Dehradun. Methods and Material: Study areas and households were selected by multistage stratified and systematic random samplings respectively. Participants were chosen from the selected households by Kish method. Respondents were screened for neuropsychiatric disorders by using MINI-6. Statistical analysis used: data was entered in SPSS 20.0 version and analysed by using percentages, chi-square test and logistic regression. Results: About one fifth of the respondents had a lifetime diagnosis of at least one neuropsychiatric disorder. Age, male gender, higher education, joint family and upper socio-economic status were found to be significantly associated with neuropsychiatric disorders. Conclusions: Factors determining psychological wellbeing are deeply rooted in socio-demographic environment and individual characteristics. Many of the respondents had one or more lifetime neuropsychiatric disorder and most of them were not diagnosed. More community-based research is required to determine the exact magnitude and responsible factors of neuropsychiatric disorders, so that a focused strategy may be developed to address its preventable aspect
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Brain microdialysis (BMD) is a technique for continuous, on-line and real-time quantitative analysis for collected samples and it has been widely used in the quantitative detection of neurotransmitters in the brain in recent years. BMD has broad prospects in the research of neuropsychiatric diseases, but it is limited in studies because of its complicated operation, high cost, long time consuming, individual difference and poor reproducibility of the probe. Optimizing experimental design and developing multilocus microdialysis are main directions of research. Monitoring the changes of the metabolic state of the brain microenvironment in the neuropsychiatric diseases is another prospect of BMD. This article reviews the principle, characteristics and key points of brain microdialysis, along with the relationship between neurotransmitters and neuropsychiatric diseases, and the progress of microdialysis in neuropsychiatric diseases.
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In recent years, a considerable number of studies have shown that N-acetylcysteine( NAC) is a promising agent in the treatment of a variety of neuropsychiatric disorders. The present article briefly outlined its role in the regulation of these disorders and reviewed the current literatures on the use of NAC in autism and obsessive-compulsive related disorders(OCRD)in order to provide ba-sis for the clinical application of NAC in neuropsychiatric field.
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Introducción: el sistema nervioso central es uno de los órganos afectados, de manera directa e indirecta, por el VIH; adicionalmente, los trastornos psiquiátricos son más frecuentes en esta población. Objetivo: comprender la patogénesis, las manifestaciones clínicas y el manejo de las enfermedades neuropsiquiátricas en la población con VIH. Métodos: se realizó una búsqueda en las bases de datos Medline, Embase, SciELO, LILACS y Psychlnfo utilizando término libres y MeSH. Resultados: los trastornos neuropsiquiátricos generan un impacto negativo en el tratamiento y seguimiento de los pacientes con VIH, lo cual disminuye su adherencia al tratamiento y aumenta las dificultades en su manejo integral. Llamativamente, existe una falta de estudios latinoamericanos en esta área.
Introduction: HIV is the infectious disease with the biggest worldwide impact in the last decades. For this reason, the control of this disease was induded in the millennium objectives of the UN. There has been a long time interest for studying the psychiatric comorbidities in these patients due to its impact in survival. Níethods: A research in the data bases Medline, Embase, SciELO, LILACS, and Psychlnfo was made using free terms and MeSH terms. Results: The neuropsychiatric disorders have a negative impact in the treatment and control of HIV positive patients, reducing their adherence and increasing the difficulties in the comprehensive treatment. There is a déficit in studies that could clear up this relationship in Colombian population.
Subject(s)
HIV , Colombia , Affective Symptoms/diagnosisABSTRACT
Neurosteroids are synthesized from cholesterol within the microglia of the central nervous system. They are classified as dehydroepiandrosterone,progesterone, pregnenolone,allopregnanolone and other peripheral steroids. Neurosteroids such as allopregnanolone are positive allosteric modulators of GABAA receptors and exert anxiolytic and antidepressant-like behavior. Neuroactive steroids play a significant role in neurodevelopment in terms of internal environment homeostasis. This paper reviews the biosynthesis, metabolism regulatory effect on the response to stress and therapeutic potentials of neurosteroids.
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This study was aimed to investigate the mechanism underlying the neuropsychiatric comorbidity in the process of "stress"-"inflammation"-"comorbidity",from the perspective of traditional Chinese medicine (TCM) basic theory in combination with our previous 20-year findings.Notably,the neural-psychiatric comorbidity between psychiatric disorders,including depression,schizophrenia and anxiety,and systemic diseases,such as ischemic stroke,Alzheimer's disease (AD) and asthma,have something in common in the pathophysiological mechanism.The stress-induced structural and functional changes in the brain,the stress-initiated diversely structural and neurobiological changes in neurocircuitry,and the stress-mediated neurochemical alterations in neurotransmitter are considered to be involved in the common pathophysiological mechanisms in the neural-psychiatric comorbidity,which initiates a cascade of physiological and psychological processes that contributes to the development of various types of neuropsychiatric disorders.Accordingly,it will be of great significance to investigate mechanisms underlying the neuropsychiatric comorbidity in the process of "stress"-"inflammation"-"comorbidity" under the guidance of the basic theory of "treating disease from the root" and "same treatment for different diseases" in TCM.
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Neurosteroids are synthesized from cholesterol within the microglia of the central nervous system. They are classi?fied as dehydroepiandrosterone,progesterone,pregnenolone,allopregnanolone and other peripheral steroids. Neurosteroids such as al?lopregnanolone are positive allosteric modulators of GABAA receptors and exert anxiolytic and antidepressant-like behavior. Neuroac?tive steroids play a significant role in neurodevelopment in terms of internal environment homeostasis. This paper reviews the biosynthe?sis,metabolism regulatory effect on the response to stress and therapeutic potentials of neurosteroids.
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Resumen:La encefalitis por anticuerpos antirreceptor N-metil-D-Aspartato pertenece a un grupo de enfermedades que inicialmente se llamaban encefalitis límbicas, hoy conocidas como encefalitis autoinmunes. Se caracterizan por estar frecuentemente asociadas a algún tumor, responder a la inmunoterapia y provocar daños cerebrales variables que se manifiestan como cuadros clínicos polimorfos (desde la disfunción límbica hasta una encefalopatía multifocal o difusa). Algunos pacientes con condiciones psiquiátricas agudas podrían tener en realidad una encefalitis autoinmune, sin embargo, no es tan fácil sospecharlo con las manifestaciones iniciales. Su incidencia exacta no se conoce, pero cada vez hay más reportes de casos. Sus características clínicas están bien descritas y evolucionan en cinco fases. El diagnóstico definitivo se hace al obtener la confirmación serológica de los anticuerpos, pero hay algunas pruebas que pueden orientar la sospecha diagnóstica. El tratamiento se basa en esteroides y otros inmunosupresores.
Abstract:Anti NMDA-receptor encephalitis belongs to a group of diseases originally called limbic encephalitis, nowadays known as autoimmune encephalitis. These encephalitis are characterized by a frequent association with a tumor, a good response to immunotherapy, and by causing a broad range of brain injuries which manifest in polymorphous clinical pictures (from limbic dysfunction to multifocal or diffuse encephalopathy). Some patients diagnosed with acute psychiatry conditions may have instead autoimmune encephalitis. Nevertheless, it is not easy to discern one from the other in the initial stages. Its exact incidence is unknown, but there are increasingly more case reports. Its clinical characteristics are well described and evolve in five stages or phases. The final diagnosis is made when antibodies serologic confirmation is obtained, but some other tests may help guide the diagnosis. The treatment is based on steroids and other immunosuppressants.
Subject(s)
Humans , Anti-N-Methyl-D-Aspartate Receptor Encephalitis/diagnosis , Encephalitis , Neuropsychiatry , Costa RicaABSTRACT
We reviewed trigeminal nerve stimulation (TNS) and transcutaneous vagus nerve stimulation (tVNS). All techniques have shown preliminary promising results, although the results are mixed. Method: We performed a systematic review of the Medline and Embase databases, with no constraint to dates, through June 2013. The keywords were [(1) trigeminal nerve stimulation OR (2) cranial nerve OR (3) trigemin* OR (4) transcutaneous VNS OR (5) transcutaneous cranial nerve stimulation] and (6) mental disorders. Results: We included four preclinical and clinical five studies on TNS. All clinical data were based on open-label studies with small samples, which diminished the external validity of the results, thus reflecting the modest impact of TNS in current clinical practice. Of the tVNS clinical trials, three assessed physiological features in healthy volunteers, and one examined patients with epilepsy. Conclusion: TNS and tVNS improve treatment of particular neuropsychiatric disorders such as depression. .
O uso de estimulação de nervos cranianos de maneira transcutânea tem sido uma estratégia em desenvolvimento recente. Diferentes estudos apontam para resultados clínicos favoráveis no tratamento de diferentes quadros neuropsiquiátricos. Método: Revisão sistemática da literatura com base nas bibliotecas eletrônicas Medline e Embase, sem restrição de data inicial, até agosto de 2013. Os termos de busca utilizados foram [(1) trigeminal nerve stimulation OR (2) cranial nerve OR (3) trigemin* OR (4) transcutaneous VNS OR (5) transcutaneous cranial nerve stimulation] and (6) mental disorders. Resultados: Incluímos quatro estudos pré-clinicos e cinco estudos clínicos abordando estimulação do nervo trigêmeo. Todos os estudos foram abertos, com pequenas amostras, o que reduz a validade externa dos dados, refletindo a ainda incipiente atuação da técnica, apesar de promissora. Considerando-se a estimulação do nervo vago, três artigos avaliaram aspectos fisiológicos em voluntários saudáveis e um artigo estudou pacientes com epilepsia. Conclusão: As estratégias de estimulação transcutânea de nervos cranianos, apesar de incipiente, tem demonstrado resultados clínicos favoráveis no tratamento de distúrbios neuropsiquiátricos. .
Subject(s)
Female , Humans , Male , Nervous System Diseases/therapy , Transcutaneous Electric Nerve Stimulation/methods , Trigeminal Nerve/physiology , Vagus Nerve Stimulation/methods , Mental Disorders/therapy , Treatment OutcomeABSTRACT
The SLC6A4 gene encodes the serotonin transporter SERT. Since the discovery of the role of SLC6A4 polymorphisms on human behavior, there is an increasingly growing wealth of information regarding SLC6A4 gene variants associated with anxiety and mood disorders, as well as their pharmacogenetic implications. In this brief review, the main discoveries on SLC6A4 variants, their functional impact and their suggested roles in neuropsychiatric and neurodevelopmental disorders are discussed.
El gen SLC6A4 codifica el transportador de serotonina SERT. Desde el descubrimiento inicial del rol que tienen polimorfismos de SLC6A4 en el comportamiento humano, hay una creciente cantidad de información acerca de variantes genéticas de SLC6A4 asociadas con trastornos de ansiedad y de estado de ánimo, así como de sus implicancias farmacogenéticas. En esta breve revisión, se discuten los principales descubrimientos de variantes de SLC6A4, su impacto funcional y sus roles sugeridos en enfermedades neuropsiquiátricas y de neurodesarrollo.
Subject(s)
Humans , Serotonin , Serotonin Plasma Membrane Transport Proteins , Nervous System DiseasesABSTRACT
γ-aminobutyric acid A(GABAA)receptor is one of the ligand-gated ion channel receptors expressing widely in the brain. As the major inhibitory neurotransmitter receptor, GABAA receptor is involved in the regulation of many physiological mechanisms. Imbalance expression of the receptor may lead to neuropsychiatric disorders such as schizophrenia,depression, epilepsy,post-traumatic stress disorder (PTSD) syndrome,neurofibromatosis type 1 and fragile X syndrome. Clinically classic benzodiazepines acting on GABAA receptor can be used for the auxiliary treatment of various neuropsychiatric disorders,which supports the potential therapeutic value of GABAA receptor for these diseases. The paper describes the molecular structure and subtypes of GABAA receptor and further reviews the progress in the link between GABAA receptor and neuropsychiatric disorders in order to provide a new target for the drug therapy development of neuropsychiatric disorders.
ABSTRACT
γ-aminobutyric acid A (GABAA) receptor is one of the ligand-gated ion channel receptors expressing widely in the brain. As the major inhibitory neurotransmitter receptor, GABAA receptor is involved in the regulation of many physiological mechanisms. Imbalance expression of the receptor may lead to neuropsychiatric disorders such as schizophrenia, depression, epilepsy, post-traumatic stress disorder (PTSD) syndrome, neurofibromatosis type 1 and fragile X syndrome. Clinically classic benzodiazepines acting on GABAA receptor can be used for the auxiliary treatment of various neuropsychiatric disorders, which supports the potential therapeutic value of GABAA receptor for these diseases. The paper describes the molecular structure and subtypes of GABAA receptor and further reviews the progress in the link between GABAA receptor and neuropsychiatric disorders in order to provide a new target for the drug therapy development of neuropsychiatric disorders.