Digital Commons Network^™

Travel Time Estimation And Prediction For Urban Arterial Roads, Porntep Puangprakhon

Chulalongkorn University Theses and Dissertations (Chula ETD)

Travel time information has been accepted as the core of advanced traveler information systems (ATIS) and advanced traffic management systems (ATMS). Providing the accurate travel time information to traffic operators and travelers allows them to make informed decisions, leading to more advantage for individual road users and the entire transportation system. Most of the traffic information providers normally deliver the current traffic conditions or current travel times to public assuming the state of traffic remains constant in the near future. Aimed at the more effective applications, short-term future traffic conditions have been proposed as a valuable piece of information in …

Prototype Development Of Small Scale Transportable Fuel Production System For Agricultural Byproducts, Jurarat Nisamaneenate

Chulalongkorn University Theses and Dissertations (Chula ETD)

The objective is to study the potential of peanut shell waste and cassava rhizome conversion using a modular fixed bed gasifier coupled with thermal integration unit. The thermal integration unit improved gasification reaction in which lower tar content and high gas production efficiency can be achieved. The air flow rate had integrated effects on product yield and composition; higher air flow rate resulted in higher gas yield with less tar and char. The result from peanut shell gasification indicated the optimal conditions without catalyst addition at air flow rate of 3.06 m3/hr where carbon and hydrogen conversions were 87.10% and …

The European Union(Eu) As A Global Actor --- Internationalisation Of Eu Fisheries Policy And Its Impact On Third Countries:Case Study Of Thailand, Ran Lu

Chulalongkorn University Theses and Dissertations (Chula ETD)

The concept of global management of marine resources, especially fisheries governance is not a radically new topic. After World War II, mainly through the UN, FAO and increasing regional institutions, regulatory and implementing mechanisms began incrementally forming. The EU, as a global actor, vertically, absorbed these universal regulations with feedback from its experimentalist governance[1] and then uploaded its “Europeanized” ideas to the international arena, meanwhile, horizontally, externalizing the EU model to third countries. This thesis will focus on the role of the EU as an international actor in the field of fisheries policy by analyzing the IUU (illegal, unreported and …

Vicariance And Oceanic Barriers Drive Contemporary Genetic Structure Of Widespread Mangrove Species Sonneratia Alba J. Sm In The Indo-West Pacific, Alison K.S Wee, Jessica Xian Hui Teo, Jasher L. Chua, Sankararamasubramanian H. Meenakshisundaram, Koji Takayama, Takeshi Asakawa, Onrizal Onrizal, Erwin Riyanto Ardli, Bayu Adjie, Sarawood Saengkaw, Monica Suleiman, Nguyen Xuan Tung, Severino G. Salmo Iii, Orlex Baylen Yllano, Mohd Nazre Saleh, Khin Khin Soe, Yoichi Tateichi, Yasuyuki Watano, Yoshiaki Tsuda, Tadashi Kajita, Edward L. Webb

Environmental Science Faculty Publications

Patterns of genetic structure are essential for a comprehensive understanding of the evolution and biogeography of a species. Here, we investigated the genetic patterns of one of the most widespread and abundant mangrove species in the Indo-West Pacific, Sonneratia alba J. Sm., in order to gain insights into the ecological and evolutionary drivers of genetic structure in mangroves. We employed 11 nuclear microsatellite loci and two chloroplast regions to genotyped 25 S. alba populations. Our objectives were to (1) assess the level of genetic diversity and its geographic distribution; and (2) determine the genetic structure of the populations. Our results …

The Search For Consistent Intonation: An Exploration And Guide For Violoncellists, Daniel Hoppe

Theses and Dissertations--Music

This paper provides a system that helps diagnose and address the specific challenge to cellists of intonation in any passage. Learning to play consistently in tune is essential for every cellist. In the over three hundred years of cello history, teachers have tried approaching the topic from a variety of perspectives. While each technique is useful in its own right, there is scant attention to how they work together. Viewing intonation through its component sub-skills is the first step toward integrating existing exercises and paving the way for further advances in pedagogy. The following paper categorizes training techniques according to …

Th1 Cells In Cancer-Associated Inflammation, Güneş Di̇nç Akbulut, Di̇dem Özkazanç, Güneş Esendağli

Turkish Journal of Biology

The immune system is not only evolved to protect the body from pathogens, but it also recognizes and eliminates cancer cells. CD4+ helper T (Th) lymphocytes are central intercessors differentiated according to the character of physiological or pathological status. Generation of type 1 Th (Th1) cells is primarily associated with a pathological insult that must be removed through immune elimination. Upon interacting with other immune and transformed cells, Th1 cells can hamper cancer progression. Therefore, it is a major obstacle for tumor cells to become insensitive or resistant to Th1-oriented actions. The organism employs various mechanisms to return to a …

Tumor Necrosis Factor-Alpha Induced Caspase-3 Activation-Related Inos Gene Expression In Adp-Activated Platelets, Özge Çevi̇k, Zelal Adigüzel, Ahmet Tarik Baykal, Azi̇ze Şener

Turkish Journal of Biology

Platelets are sensitive cells and are easily activated by different stimulants in the circulation system. It is known that tumor necrosis factor-alpha (TNF-α) is a proinflammatory cytokine and plays a role in inflammation. The role of TNF-α in the apoptotic process in blood platelets is unknown. In order to study the formation of apoptosis in platelets after incubation with TNF-α and/or ADP, several biomarkers were chosen: phosphatidylserine (PS) exposure and P-selectin binding; cGMP, Cyt-c, and Ca2+ levels and NOS activation; and gene and protein expression of caspase-3 and iNOS. Platelets were incubated with 100 pg/mL TNF-α and/or 50 mM iNOS …

Expansion Of Human Umbilical Cord Blood Hematopoieticprogenitors With Cord Vein Pericytes, Betül Çelebi̇ Saltik, Beyza Gökçinar Yağci

Turkish Journal of Biology

The vascular niche is a site rich in blood vessels, whereas endothelial cells, pericytes, and smooth muscle cells create a microenvironment that recruits mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs), which is important for stem cell mobilization, proliferation, and differentiation. In this study, CD146 + pericytes were purified and enriched from the human umbilical cord vein. In order to define their direct role in hematopoiesis, we tested the CD146 + pericytes as compared with osteoblasts derived from umbilical cord blood (UCB) MSCs to sustain human UCB hematopoietic progenitor cells in noncontact coculture settings or in culture media previously …

Assessment Of The Function And Expression Pattern Of Auxin Response Factor B3 In The Model Legume Plant Medicago Truncatula, Miglena Revalska, Valya Vassileva, Grigor Zehirov, Sofie Goormachtig, Anelia Iantcheva

Turkish Journal of Biology

The phytohormone auxin is a critical signal molecule, regulating fundamental processes in plant growth and development, such as shaping the root and shoot architecture, organ patterning, and nodulation. Auxin regulates plant gene expression mainly through auxin response factors (ARFs), which bind to auxin response elements in the promoter, upstream of auxin-activated genes. Here we examine and assess the function and expression pattern of a gene described as an auxin response factor, containing a DNA-binding pseudobarrel and B3 DNA-binding domains, from Medicago truncatula (MtARF-B3). For the model legume species M. truncatula, stable transgenic plants with MtARF-B3 overexpression, downregulation, and transcriptional reporters …

Stably Expressed Reference Genes During Differentiation Ofbone Marrow-Derived Mesenchymal Stromal Cells, Ilgin Cagnan, Fatima Aerts Kaya, Fahri̇ye Duygu Çeti̇nkaya, Ayşen Özcan

Turkish Journal of Biology

Human mesenchymal stromal cells (MSCs) from different parts of the body (i.e. bone marrow, BM) have distinct cellular and molecular features including global gene expression profiles. Quantitative polymerase chain reaction is a reliable method used in the quantification of gene expression. Correct assessment of target gene expression mostly depends on the reference gene (RG) of choice. Herein, expression levels of RGs (n = 19) in adipo-/osteogenic-differentiated and control (uninduced) BM-MSCs from donors were measured using the RealTime ready Human Reference Gene Panel. Characterization of BM-MSCs was assessed using in vitro differentiation by histochemical staining and immunophenotyping of cells by flow …

Expression Of The Zmdef1 Gene And α-Amylase Inhibitory Activityof Recombinant Defensin Against Maize Weevils, Thi Xuan Thuy Vi, Hoang Duc Le, Vu Thanh Thanh Nguyen, Van Son Le, Hoang Mau Chu

Turkish Journal of Biology

Plant defensins are multifunctional small cysteine-rich proteins. They are active against fungi, bacteria, and many viruses, and they inhibit trypsin and α-amylase activities. In this study, we expressed the maize defensin gene (ZmDEF1) in tobacco seeds in order to establish the basis for generating transgenic maize plants resistant to weevils. The ZmDEF1 gene was isolated from Maison, a Vietnamese local maize cultivar, which is well known for having the highest resistance to weevils among other local cultivars. The ZmDEF1 gene was cloned into a binary vector, pBetaPhaso-dest, which carries phaseolin, a seed-specific promoter, to direct defensin expression in tobacco seeds. …

Cloning, Expression, And Characterization Of Human Brain Acetylcholinesterase In Escherichia Coli Using A Sumo Fusion Tag, Hami̇d Ceylan, Orhan Erdoğan

Turkish Journal of Biology

The molecular structure of acetylcholinesterase (AChE) attracts interest because of its versatility and significant role in the cholinergic system. The main purpose of the present study was to clone a full-length cDNA sequence of human brain acetylcholinesterase (hAChE) into pET SUMO vector and express it successfully. The integrity of the constructed plasmid was confirmed by cross PCR. This recombinant construct was expressed in Escherichia coli BL21 (DE-3). In this work, we produced hexahistidine (6xHis) tagged fusion protein by isopropyl β-D-1-thiogalactopyranoside (IPTG) induction and purified using nickel (Ni2+) affinity chromatography. Using anti-His antibody, we detected ~90 kDa fusion protein. The expression …

Alteration In The Subcellular Location Of The Inhibitor Of Growth Proteinp33(Ing1b) In Estrogen Receptor Alpha Positive Breast Carcinoma Cells, İmge Kunter, Emi̇ne Kandemi̇ş, Hani̇ Alotai̇bi̇, Tülay Canda, Şeri̇fe Esra Erdal Bağriyanik

Turkish Journal of Biology

ING1 has regulatory roles in the expression of genes associated with proliferation, apoptosis, and senescence. p33(ING1b) is the most widely expressed isoform of the gene. Downregulation of its nuclear expression is involved in differentiation and pathogenesis in invasive breast carcinoma. Yet the mechanism(s) by which p33 nuclear targeting is regulated remains unknown. In this study, we analyzed human invasive breast carcinoma tissue samples by immunostaining with p33 and correlating p33 location with the presence of ERα. Our findings show the expression of p33 protein in ERα-positive tumor samples was in the nucleus alone, while the expression was mainly in the …

Studies On Organogenesis During Regeneration In The Earthworm, Eudrilus Eugeniae, In Support Of Symbiotic Association With Bacillus Endophyticus, Elaiya Raja Subramanian, Dineshkumar Sudalaimani, Johnson Retnaraj Samuel Selvan Christyraj, Kalidas Ramamoorthy M, Nino Gopi D, Jackson Durairaj Selvan Christyraj, Kasimani Renganathan, Sundar Krishnan, Sudhakar Sivasubramaniam

Turkish Journal of Biology

Organogenesis and animal adaptation upon loss of body parts are crucial events in regeneration of all traits, and both are uncovered areas in biology. The organogenesis of the mouth and anus during regeneration was studied in Eudrilus eugeniae. It was found that the earthworm regenerates a functional mouth and anus in six days. During the course of organogenesis, the earthworm adopts unique starvation behavior for 5 days, and the starvation triggers the gut microflora of E. eugeniae to produce riboflavin. One of the efficient riboflavin-producing bacteria, Bacillus endophyticus, confirmed through 16S rRNA sequencing, has been isolated from its gut. The …

Lipocalin Gene Expression Is Varied In Developmental Stages By Larval Nutritional Stress In Drosophila, Nazli Ayhan, Pinar Güler, Banu Şebnem Önder

Turkish Journal of Biology

Lipocalin genes NLaz, GLaz, and Karl are evolutionarily conserved genes in Drosophila melanogaster. There are studies on lipocalin gene expression differences under diverse diet conditions, but these studies have focused mainly on age-dependent expression profiles of these genes. The main aim of our study is to determine lipocalin expression in the developmental period by nutritional manipulation with an isofemale-based design. Three larval developmental periods have been researched under normal and restricted diets. We found significant differences between lines during their developmental time-related lipocalin expression. Here, we demonstrate that upregulations in the early developmental stages of lipocalin genes under stressful conditions …

Cryopreservation Of Citrus Seed Via Dehydration Followedby Immersion In Liquid Nitrogen, Ergun Kaya, Fernanda Souza, Emel Yilmaz Gökdoğan, Muammer Ceylan, Maria Jenderek

Turkish Journal of Biology

An important method for plant germplasm conservation is offered by a biotechnology-based approach of cryopreservation. Cryopreservation refers to the storage of plant material at ultralow temperatures in liquid nitrogen. A procedure for cryopreservation of polyembryonic seeds was improved for selected citrus cultivars from Turkey. Seed dehydration was performed at different exposure times, in sterile conditions of a laminar flow-hood. The tested cultivars showed the highest tolerance to low temperature storage when the seeds were first dehydrated to a moisture content of 21.8% for Poncirus trifoliata Raf. x Citrus sinensis Osb. and to 17.6% for Citrus limonia Osbeck. The postcryopreservation germinability …

Hyper-Activation Of Pp60(Src) Limits Nitric Oxide Signaling By Increasing Asymmetric Dimethylarginine Levels During Acute Lung Injury, Sanjiv Kumar, Xutong Sun, Satish Kumar Noonepalle, Qing Lu, Evgeny Zemskov, Ting Wang, Saurabh Aggarwal, Christine Gross, Shruti Sharma, Ankit A. Sesai, John D. Catravas

Bioelectrics Publications

The molecular mechanisms by which the endothelial barrier becomes compromised during lipopolysaccharide (LPS) mediated acute lung injury (ALI) are still unresolved. We have previously reported that the disruption of the endothelial barrier is due, at least in part, to the uncoupling of endothelial nitric oxide synthase (eNOS) and increased peroxynitrite-mediated nitration of RhoA. The purpose of this study was to elucidate the molecular mechanisms by which LPS induces eNOS uncoupling during ALI. Exposure of pulmonary endothelial cells (PAEC) to LPS increased pp60^Src activity and this correlated with an increase in nitric oxide (NO) production, but also an increase in …

A Model Of Distress Tolerance In Self-Damaging Behaviors: Examining The Role Of Emotional Reactivity And Learned Helplessness, Brittany Kay Sommers

Dissertations

Problem

Although the literature is clear that low emotional distress tolerance is associated with a myriad of self-damaging behaviors, very little is known about individual difference factors in distress tolerance. Both theoretical and empirical support suggest that emotional reactivity and learned helplessness may be individual difference factors in distress tolerance. Specifically, individuals with high emotional reactivity and high learned helplessness may be at risk for low distress tolerance. Further research was needed to clarify the role of emotional reactivity and learned helplessness in distress tolerance in the context of self-damaging behaviors.

Method

Participants completed surveys which measured their (a) emotional …

Emotional Intelligence And Decision-Making As Predictors Of Antisocial Behavior, Melina Sample

Dissertations

Problem

Antisocial behaviors in the general population are not well understood and little studied, however, their acceptance is becoming more normalized and accepted by society. This present study intended to focus on the contributing factors of emotional intelligence and decision-making to the presence of antisocial behaviors, and how these contributions vary by gender.

Method

This study used the Levenson Self-Report Psychopathy Scale, the Trait Emotional Intelligence Questionnaire – Short Form, and the General Decision Making Style Inventory to explore the relationships between the three variables. A structural equation model was developed to examine the theoretical covariance matrix proposed in comparison …

Does The Inverse Of Bergmann’S Rule Apply To The Hemidactylus Genus?, Kristie I. Armas

USF St. Petersburg campus Honors Program Theses (Undergraduate)

The applications of Bergmann’s rule on nocturnal squamates has rarely been examined. Bergmann’s rule states that species increase in size with increasing latitude. The relationship between latitudinal gradients and body size may shed light on the mechanisms these geckos use to function under suboptimal temperatures. This study examined maximum body size in relationship to absolute latitude in 33 of the 132 Hemidactylus species. The objective of the study was to test the inverse of Bergmann’s rule, thus testing if lizard body sizes decrease towards higher latitudes. Data was collected from literature and the following two databases, Vertnet and The Reptile …

Impacts Of Bunces Pass Boat Traffic On The Marine Environment, Tiffany Nicholson

USF St. Petersburg campus Honors Program Theses (Undergraduate)

Each year nearly 1.4 billion pounds of trash enter the ocean from either a point source or runoff from the land (NOAA 2011). Yet aside from the land pollution that is produced, there is also a large amount of water pollution that takes place each year. This includes the fishing industry, shipping industry, oil spills, and one of the most underrated one is anthropogenic noise pollution. In 2010, NOAA reported that roughly 39% of America’s population are living in counties directly on the shoreline with an expected increase of an additional 8% by 2020 (NOAA 2010). With more people moving …

Mindset, Academic Motivation, And Academic Self-Efficacy As Correlates Of Academic Achievement Among Undergraduate Students In Communication Sciences And Disorders Programs, Heather Loraine Ferguson

Dissertations

Problem

Higher education serves many stakeholders including students, parents, faculty, staff, university administrators, and other contributors. Those stakeholders are all linked with one purpose: the success of the student. That success or failure is most commonly measured by achievement through grade point average (GPA). The academic demands within the college/university setting is high. Limited academic achievement can result in academic failure, being placed on academic probation, even losing scholarships and grants. Students enter college, progress through college, and often graduate without having a real understanding for what it truly takes to be academically successful and what factors may contribute to …

Impact Of Concussion On Youth: Pediatric Perspectives Of Recovery Related To Learning, Emotional Health, Social Life, And School, Ruta Clair

PCOM Psychology Dissertations

Pediatric concussion is a significant health concern for parents, medical providers, and schools. There is little research about how children and adolescents view their recovery and the challenges that they face while their brains gradually restore metabolic function. This study was designed to gain insight into the perspectives of children and adolescents recovering from concussion. Further, exploration of factors, such as type and intensity of symptoms, time since injury, gender, and age of patient, were examined to provide an understanding of the interaction between these factors and the impact on the individual’s perception of his or her recovery. The Pediatric …

Exploring The Relationships Between Pornography Consumption, Relationship Satisfaction, Relationship Beliefs, And Masculinity, Benjamin Ryan Barnes Jr.

PCOM Psychology Dissertations

Pornography is widely available and consumed in the United States and worldwide. Despite pornography’s wide spread use and expansions since the advent of the Internet, the psychosocial problems related to its use are not fully delineated. Previous studies have explored the relationship between pornography consumption and psychosocial variables such as sexual and physical violence against women, misogynistic beliefs, sexual callousness, acceptance of rape myths, and other problematic sexual beliefs and behaviors. Fewer studies have explored pornography use and its impact on relationship variables such as relationship satisfaction and beliefs about relational constructs such as intimacy, passion, and equality/independence. Furthermore, there …

Effect Of Calprotectin Subunit S100a9 On The Expression And Methylation Of Ocln In Human Melanoma Cell Line A-375, Mostafa Najafi, Safar Ali Alizadeh, Mehdi Azad, Taghi Naserpour Farivar, Farzad Rajaei, Kourosh Hotam Sorouri, Babak Rahmani, Nematollah Gheibi

Turkish Journal of Biology

Increased levels of calprotectin subunits S100A8 and S100A9 have been detected in human cancers. Melanoma is the most aggressive type of skin cancer, and its treatment is challenging because of its brain metastasis. OCLN encodes occludin, which plays a major role in the formation and regulation of tight junctions. The aim of this study was to evaluate the methylation status of the OCLN promoter and its expression in A-375 melanoma cells treated with or without various concentrations of S100A9 for 24, 48, and 72 h. Total RNA was extracted, and synthesized cDNA was assessed by performing real-time PCR. MSP-PCR was …

Synergistic Effect Of 24-Epibrassinolide And Salicylic Acid On Photosynthetic Efficiency And Gene Expression In Brassica Juncea L. Under Pb Stress, Sukhmeen Kaur Kohli, Neha Handa, Anket Sharma, Vinod Kumar, Parminder Kaur, Renu Bhardwaj

Turkish Journal of Biology

Lead (Pb) has wide-ranging effects on various essential physiological processes in plants including seed germination, root/shoot growth, photosynthetic efficiency, water status, and activities of enzymes. The effect of combined treatment of 24-epibrassinolide (10-7 M, EBL) and salicylic acid (1 mM, SA) on growth, photosynthetic attributes, and phenolic compounds in 30-, 60-, and 90-day-old plants of Brassica juncea L. under Pb stress (0.25 mM, 0.50 mM, and 0.75 mM) were studied. It was observed that Pb toxicity resulted in lowered growth and photosynthetic efficiency. The expressions of CHLASE, PSY, CHS, and PAL genes were altered. Presoaking treatment with the combination of …

The Mtt Viability Assay Yields Strikingly False-Positive Viabilities Although The Cells Are Killed By Some Plant Extracts, Di̇dem Karakaş, Ferda Ari, Engi̇n Ulukaya

Turkish Journal of Biology

The MTT assay is one of the often used cell viability/cytotoxicity assays. However, when the methanol extracts of plants are used to test their cytotoxic potential, interference may occur, resulting in false-positive viability results. Therefore, in this study, the reliability of the MTT assay was investigated in the case of plant use. The methanol extracts of three different plants (Hypericum adenotrichum, Salvia kronenburgii, and Pelargonium quercetorum) were tested in breast cancer cell lines (MCF-7 and MDA-MB-231) using the MTT assay and the results were compared to the ATP assay, which is a much more sensitive and reliable assay due to …

Melatonin Diminishes Oxidative Damage In Sperm Cells,Improving Assisted Reproductive Techniques, Fabián Monllor, Javier Espino, Ana María Marchena, Águeda Ortiz, Graciela Lozano, Juan Francisco García, Jose A. Pariente, Ana B. Rodriguez, Ignacio Bejarano

Turkish Journal of Biology

Sperm preparation procedures are a potential generator of oxidative stress-induced DNA damage, which leads to a dramatic drop in fertility. An increasing number of studies suggest that melatonin reduces the oxidative stress induced by manipulation. However, very little is known about the preservative role of melatonin in sperm preparation medium during assisted reproduction procedures. For this aim to be achieved, semen was divided into two fractions and preincubated with and without 1 mM melatonin. Afterwards, both fractions were divided into two subfractions to perform swim-up in the presence and absence of 1 mM melatonin. Labeling with anti-CD46 and antiactive caspase-3 …

Mg Deficiency Changes The Isoenzyme Pattern Of Reactive Oxygen Species-Relatedenzymes And Regulates Nadph-Oxidase-Mediated Ros Signaling In Cotton, Rengi̇n Özgür Uzi̇lday, Bariş Uzi̇lday, Tolga Yalçinkaya, İsmai̇l Türkan

Turkish Journal of Biology

The aim of this work was to investigate changes in isoenzyme patterns of enzymes related to reactive oxygen species (ROS) detoxification such as superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductase (GR), and glutathione-S-transferase (GST) in cotton under Mg deficiency. Moreover, we aimed to elucidate how a ROS producer, NADPH oxidase (NOX), responds to changing Mg levels. Cotton plants were grown with different concentrations of MgSO4 in hydroponic medium to create nutrient deficiency (0, 75, 150, 1000 μM Mg). Gradual decreases in growth and photosynthetic rates were observed with declining Mg concentrations and 0, 75, and …

Mesoporous Silica Nanoparticles Sba-15 Loaded With Emodin Upregulate The Antioxidative Defense Of Euproctis Chrysorrhoea (L.) Larvae, Milena Jankovic-Tomanic, Dajana Todorovic, Zoran Stanivukovic, Vesna Peric Mataruga, Ludger Wessjohann, Goran Kaludjerovic

Turkish Journal of Biology

The study presented here aims to elucidate the effects of emodin (EO = 1,3,8-trihydroxy-6-methylanthraquinone) in its free form and when loaded into a mesoporous silica nanocarrier SBA-15 (→ SBA-15 EO) on the activities of the main antioxidative enzymes, superoxide dismutase, catalase, glutathione S-transferase, and glutathione reductase, in larvae of a polyphagous insect pest, the browntail moth Euproctis chrysorrhoea (L.). The results show that only SBA-15 EO upregulates the activities of the tested antioxidative enzymes. These results point to significant differences in the effectiveness of the compound in the free versus the loaded form.