The authors express their extreme appreciation to the Regional&#160;Coordinating&#160;Office for Education of Rio&#160;Grande&#160;do&#160;Sul, for authorizing and granting us access to the school population. We also thank the help we received from the schools&#8217; principals and from all of the faculty members who accompanied and facilitated access to the pupils, as well as their contribution to explaining the study to the parents. Finally, we appreciate the feedback and support received from the Ethical&#160;Committees of the University&#160;of&#160;Unipampa.

The research obtained authorization from the Research&#160;Ethics&#160;Committee of the University&#160;of&#160;Unipampa. During the whole process of data collection, children&#8217;s anonymity was guaranteed and the ethical principles for medical research in human beings described in the latest revision of the Declaration&#160;of&#160;Helsinki were applied23.
1. Design of the research instrument
<ol>
	<li>Design a first draft of the questionnaire to meet the objective of the study (Supplementary File&#160;1). This questionnaire should contain questions about breakfast habits in schoolchildren. The objective is to know the relationship between breakfast habits and the sociodemographic characteristics of this population. 
	NOTE: A transcultural adaptation of the abridged questionnaire on Eating&#160;Habits of the School&#160;Population of the &#8220;PERSEO&#8221;&#160;program is conducted, validated in the Spanish population24. The items related to breakfast are selected.</li>
	<li>Create the draft using word editing software so that it can be easily shared and modified. Include different study variables such as the following: gender, age, family situation, parent&#8217;s occupation25. Variables on the breakfast habits include foods included in the daily breakfast, classification of breakfast (both according to the Spanish&#160;Society of Community&#160;Nutrition and to the recommendations of the&#160;WHO)26,27.</li>
	<li>Send a draft to external experts. Have them review the translated questionnaire whilst considering aspects such as item comprehension. 
	NOTE: The experts were a teacher and psychologist familiarized with the use and translation of questionnaires. Both people were bilingual in Portuguese and Spanish.</li>
	<li>Design a final version of the questionnaire (Supplementary File&#160;2) and submit it to a scientific and ethical committee along with a research report of the project. 
	NOTE: Positive evaluation was obtained both in scientific and ethical areas from the University&#160;of&#160;Unipampa committee (Supplementary File&#160;3).</li>
	<li>Conduct a reliability analysis of the resulting questionnaire applying it at two&#160;different moments in time in order to analyze its consistency and reproducibility in a pilot group.</li>
</ol>
2. Sampling method
<ol>
	<li>Conduct a cross-sectional descriptive study to characterize the breakfast habits of the schoolchildren.
	<ol>
		<li>Have the teacher ask for silence in the classroom.</li>
		<li>Have each student complete their questionnaire individually. They have fifteen minutes to complete it, approximately one minute per question.</li>
		<li>Ask students to be honest with their answers and select only one option for each question. 
		NOTE: A random sampling was performed by using clusters of&#160;urban public schools of the city (Figure 1). In the random selection of clusters, 12&#160;schools were included with a total of 558 boys and girls. To reach an acceptable confidence level of 95% and marginal error of 5%, the minimum sampling size should be 264 children.</li>
	</ol>
	</li>
	<li>Contact the leaderships of all the participating schools (directors or heads of studies) to report the date and time for completing the questionnaires.</li>
	<li>Inform parents and obtain their informed consent. Children complete questionnaires individually but the consent of their patents is essential because they are minors.</li>
</ol>
3. Statistical analyses
<ol>
	<li>Analyze the data with a statistical analysis program (e.g., SPSS). Manually pass all written data to a database in a spreadsheet format. This format allows opening database in the statistical program. Analyze all parameters completed by the students. Select Analyze | Descriptive Statistics | Crosstabs | Statistics | Kappa | Accept (see Figure 2). 
	NOTE: To evaluate the reliability and reproducibility of the transcultural adaptation of the questionnaire, the Kappa indexes of concordance of the dichotomic questionnaire items are calculated. For the polichotomic items, the summatory value of the scores obtained is used, generating a new variable called &#8220;sum of the items&#8217; scores&#8221;, with values from&#160;0&#160;to&#160;4. The interclass correlation coefficient is calculated to assess the numerical concordance between the first and the second measurements. Both for the Kappa index and for the aforementioned coefficient, values higher than or equal to&#160;0.40 are considered acceptable, and those higher than or equal to&#160;0.75 are considered excellent28. The accepted statistical significance level to calculate these coefficients was set at p&#160;&#60;&#160;0.05.</li>
	<li>Perform the descriptive analysis: calculate the frequency distributions of the main qualitative variables, as well as the measurements of central tendency and dispersion of the quantitative variables. Select Analyze | Descriptive Statistics | Explore | Confidence interval for the mean 95% | Accept (see Figure 3).</li>
	<li>Perform hypothesis contrast by using the Chi-square&#160;test and Student&#8217;s&#160;t-test, depending on the case. For the Chi-square test, select Analyze | Descriptive Statistics | Crosstabs | Statistics | Chi-square (see Figure 4). For Student&#8217;s t-test click Analyze | Compare averages | T test for independent samples (see Figure 5). 
	NOTE: The accepted statistical significance level for the hypothesis contrast is set at&#160;p&#160;&#60;&#160;0.05.</li>
</ol>

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	<li>Ogden, C. L., Carroll, M. D., Kit, B. K., Flegal, K. 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The authors have nothing to disclose.

This study describes the profiles of schoolchildren with regards their breakfast eating habits. It allows us to approach the possible factors that are involved in omitting breakfast. These data provide knowledge of the population to conduct a future health educational program in this collective group.
One of the limitations of the study was the fact that the questionnaire is not validated in the Portuguese language, but in Spanish. For this reason, a transcultural adaptation was conducted into Portuguese, and its reliability was analyzed in two&#160;grades and in two&#160;different times of the same week, in order to analyze the consistency and reproducibility of the questionnaire. This was a critical step in the protocol. The sample was exhaustively selected by clusters and randomization was guaranteed. This is a representative sample, since the questionnaires were applied in 12&#160;different schools, from the total of&#160;24 which make up the city of&#160;Uruguaiana. The response rate was high.
Our study showed a statistically significant association between the breakfast habits and the frequency of having breakfast with the parents; data which coincide with other authors who assert the relevance of the parents having breakfast with their children, and even having them help prepare the meal. Fugas&#160;et&#160;al observed that one of the main reasons for omitting breakfast was lack of time, and found a significant difference between the weekly frequency of having breakfast and it being prepared by a family member19. As regards gender, others authors29,30 found that the habit of having breakfast was more developed among the boys&#160;(95.1%) than among the girls&#160;(92.5%), a fact which differs from our study, since the percentage of girls who had breakfast was higher (73.7%&#160;in boys, 79.5%&#160;in girls), although no significant difference was found in this parameter.
In our study, only 18% state they have breakfast every day of the week, in opposition to other studies conducted in Canada31, Spain4 and Argentina19, where 85.5%, 77.5%, and 75%, respectively, have breakfast at their homes every day. In the study conducted in Spain, specifically in Andalusia, 5.2%&#160;of the pupils eat fruits for breakfast4, in opposition to 24.4% in our study. In the study conducted in Argentina19, 16% eat a low quality breakfast based on candies and industrial pastries and, in another study conducted in Italy32, the percentage rises to 31.3% of children eating this type of foods for breakfast. These data differ from our study, where only 3.1% have this kind of breakfast. In our study, the children have breakfasts mainly based on milk, eggs, bread, dried fruits, fresh fruits or natural fruit juices.
In conclusion, we can observe that despite the fact that in our population the breakfast habit is deficient; it is observed that the foods eaten are healthier than those referred to in other studies conducted in different countries. For example, they consume more fruits and less candies, pastries and salted snacks. There are diverse sociodemographic parameters which have not shown any statistical significance with the breakfast habit; this motivates us to conduct new studies including new variables. It would even be interesting to conduct a qualitative study to explore the reasons for omitting breakfast.
As future application, this study is yet another justification to launch educational programs for the health of schoolchildren, also including their parents, in order to attain better eating behaviors and knowledge on the importance of breakfast.

Obesity, as a consequence of unhealthy nutrition and of a sedentary lifestyle, represents a priority in global public health1,2. There are yet other factors which indirectly affect obesity, like economic, social and educational determinants3. For this reason, the need arises for health promotion interventions in the field of healthy nutrition, the school age being the most propitious stage, since the habits adopted in childhood usually perpetuate into adulthood4.
Breakfast is the first and most important meal of the day; according to its content, it must guarantee an average of 25% of the total energy consumed during the day to cover the nutritional needs of the organism. A healthy breakfast is considered to contain a balanced serving of carbohydrates, lipids, proteins, vitamins, and minerals. The triad made up by dairy products, cereals and fruits are preferably advisable, which can be supplemented with protein-rich foods5. Breakfast is directly linked to positive short- and long- term results, because it is associated with the improvement in the cognitive function and in the academic performance, school attendance, and healthy growth of children and adolescents, as long as it is eaten correctly6. Despite its importance, omitting breakfast is a common practice among children and adolescents7. Diverse studies show that omitting breakfast is linked to a higher increased probability of suffering from overweight and obesity8,9,10,11. In view of that, identifying those factors linked to overweight and obesity in children by analyzing their breakfast habits is important to design more effective and personalized interventions7.
Changing the eating habits is a strategy for health promotion and disease prevention12. The eating habits constituted early in life depend on factors which include the family, the school setting, and the influence of the mass media13,14. Previous studies that explored the association between the socioeconomic condition and the breakfast habits signal that a child who lives with a monoparental family or in a low-income family is more prone to presenting breakfast irregularities15,16. In this context, the first results of the ENERGY&#160;project showed that the mean frequency of the weekly breakfast consumption in European children was 5.9&#160;days/week, from 5.1&#160;in Slovenia to 6.7&#160;in Spain17, with no significant differences between boys and girls18.
In Latin&#160;America, some studies have been conducted on the eating habits of schoolchildren, specifically addressing breakfast. In a study conducted in Santa&#160;Fe,&#160;Argentina, in which 75% of the schoolchildren stated they had breakfast daily, it was concluded that the weekly frequency of having breakfast depended on the fact that a family member prepared it for them, and one of the main factors associated to omitting breakfast was lack of time19. In another study conducted in Montevideo,&#160;Uruguay, 30% of the schoolchildren admitted that having a good breakfast was important to stay healthy20.
In Brazil, malnutrition is relevant especially in children who live with low-income families21. The children and adolescents consumed a diet high in fat and sugar and low in polyunsaturated fatty acids, fiber, iron, and calcium22. At present, the studies on food consumption habits in Brazilian children are scarce. The objective of this study is to know the diet pattern of the school population of Uruguaiana&#160;(RS,&#160;Brazil) by studying breakfast as a reference meal, and to identify the sociodemographic and family factors which can be associated with a deficient breakfast. The hypotheses of the study are that some factors such as having breakfast with parents and work situation of the parents influence the breakfast habits of children.

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breakfast habits among schoolchildren in the city of uruguaiana, brazil

Breakfast is the first and most important meal of the day, and omitting it is associated with a greater probability of overweight and school absenteeism. The eating habits constituted in children depend on social, educational and economic factors, and they usually perpetuate into adulthood. For this reason, the most propitious stage for health promotion interventions on healthy nutrition is childhood. Specifically, in Brazil, malnutrition is relevant in children from low-income families although, currently, there are few studies that relate nutrition with the social factors. In this study, a reliability analysis of the Eating&#160;Habits of the School&#160;Population questionnaire was conducted, and the items referring to the breakfast habits and to some sociodemographic variables were selected. Subsequently, a total of 470&#160;fourth&#160;grade pupils self-completed the questionnaire in 12&#160;schools in Uruguaiana&#160;(Brazil). More than 50% of the mothers and more than 70% of the fathers worked in diverse areas of services and sales in stores or supermarkets. With regards to the hypothesis contrast, a significant relation was observed between the frequency of having breakfast with the father or the mother and having had breakfast&#160;(p&#160;&#60;&#160;0.001). This type of study favors a population analysis for the subsequent design and execution of health promotion activities.

Watch this Scientific Journal Video about Breakfast Habits among Schoolchildren in the City of Uruguaiana, Brazil at JoVE.com

Breakfast Habits among Schoolchildren in the City of Uruguaiana, Brazil

This article describes the breakfast habits and the factors related to schoolchildren in a city of Brazil&#160;(Uruguaiana). To this end, a transcultural adaptation was performed of a questionnaire validated in Spain, and 470&#160;boys and girls from twelve&#160;schools were interviewed.

Breakfast is the first and most important meal of the day, and omitting it is associated with a greater probability of overweight and school absenteeism. ...

breakfast-habits-among-schoolchildren-in-the-city-of-uruguaiana-brazil

Nanyang Technological University

Research

JoVE Journal

Behavior

4.4K Views.  Federal University of Pampa. This article describes the breakfast habits and the factors related to schoolchildren in a city of Brazil (Uruguaiana). To this end, a transcultural adaptation was performed of a questionnaire validated in Spain, and 470 boys and girls from twelve schools were interviewed.

Video: Breakfast Habits among Schoolchildren in the City of Uruguaiana, Brazil

A Treatment Package without Escape Extinction to Address Food Selectivity

Feeding difficulties and feeding disorders are a commonly occurring problem for young children, particularly children with developmental delays including autism. Behavior analytic interventions for the treatment of feeding difficulties oftentimes include escape extinction as a primary component of treatment. The use of escape extinction, while effective, may be problematic as it is also associated with the emergence of challenging behavior (e.g., extinction burst). Such challenging behavior may be an acceptable side effect in treatment cases where feeding problems are severe and chronic (e.g., failure to thrive). However, in more acute cases (e.g., selective eating), the negative side effect may be unwarranted and undesired. More recent research on the behavioral treatment of food selectivity has begun to evaluate treatments for feeding difficulties that do not include escape extinction (e.g., demand fading, behavioral momentum), with some success. However, research to date reveals individual differences in responsiveness to such treatments and no clear preferable treatment has emerged. This manuscript describes a multi-component treatment package that includes shaping, sequential presentation and simultaneous presentation, for the treatment of food selectivity in four young children with developmental delays. This treatment package extends the literature on the behavioral treatment for food selectivity and offers a multi-component treatment protocol that may be clinically applicable across a range of treatment scenarios and settings.

Feeding difficulties are a common problem for children with developmental disorders, including autism, and behavioral interventions often include escape extinction. Recent research has begun to evaluate treatments that do not include escape extinction. This manuscript describes a multicomponent treatment package that does not use escape extinction to treat feeding difficulties.

Feeding difficulties and feeding disorders are a commonly occurring problem for young children, particularly children with developmental delays including ...

Determining Soil-transmitted Helminth Infection Status and Physical Fitness of School-aged Children

Soil-transmitted helminth (STH) infections are common. Indeed, more than 1 billion people are affected, mainly in the developing world where poverty prevails and hygiene behavior, water supply, and sanitation are often deficient1,2. Ascaris lumbricoides, Trichuris trichiura, and the two hookworm species, Ancylostoma duodenale and Necator americanus, are the most prevalent STHs3. The estimated global burden due to hookworm disease, ascariasis, and trichuriasis is 22.1, 10.5, and 6.4 million disability-adjusted life years (DALYs), respectively4. Furthermore, an estimated 30-100 million people are infected with Strongyloides stercoralis, the most neglected STH species of global significance which arguably also causes a considerable public health impact5,6. Multiple-species infections (i.e., different STHs harbored in a single individual) are common, and infections have been linked to lowered productivity and thus economic outlook of developing countries1,3.
For the diagnosis of common STHs, the World Health Organization (WHO) recommends the Kato-Katz technique7,8, which is a relatively straightforward method for determining the prevalence and intensity of such infections. It facilitates the detection of parasite eggs that infected subjects pass in their feces.
With regard to the diagnosis of S.stercoralis, there is currently no simple and accurate tool available. The Baermann technique is the most widely employed method for its diagnosis. The principle behind the Baermann technique is that active S.stercoralis larvae migrate out of an illuminated fresh fecal sample as the larvae are phototactic9. It requires less sophisticated laboratory materials and is less time consuming than culture and immunological methods5.
Morbidities associated with STH infections range from acute but common symptoms, such as abdominal pain, diarrhea, and pruritus, to chronic symptoms, such as anemia, under- and malnutrition, and cognitive impairment10. Since the symptoms are generally unspecific and subtle, they often go unnoticed, are considered a normal condition by affected individuals, or are treated as symptoms of other diseases that might be more common in a given setting. Hence, it is conceivable that the true burden of STH infections is underestimated by assessment tools relying on self-declared signs and symptoms as is usually the case in population-based surveys.
In the late 1980s and early 1990s, Stephenson and colleagues highlighted the possibility of STH infections lowering the physical fitness of boys aged 6-12 years11,12. This line of scientific inquiry gained new momentum recently13,14,15. The 20-meter (m) shuttle run test was developed and validated by L&eacute;ger et al.16 and is used worldwide to measure the aerobic fitness of children17. The test is easy to standardize and can be performed wherever a 20-m long and flat running course and an audio source are available, making its use attractive in resource-constrained settings13. To facilitate and standardize attempts at assessing whether STH infections have an effect on the physical fitness of school-aged children, we present methodologies that diagnose STH infections or measure physical fitness that are simple to execute and yet, provide accurate and reproducible outcomes. This will help to generate new evidence regarding the health impact of STH infections.

Chronic infection with soil-transmitted helminths (STHs) causes malabsorption, stunting, and wasting in the growing child. Hence, it is plausible that these infections also reduce the physical fitness of children. Here, we visualize two techniques for the diagnosis of STHs and the 20-meter shuttle run test for assessing children's physical fitness.

Soil-transmitted helminth (STH) infections are common. Indeed, more than 1 billion people are affected, mainly in the developing world where poverty ...

Immunology and Infection

Assessment of Social Transmission of Food Preferences Behaviors

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy aging groups. For example, olfactory dysfunction in AD can present as impairment in olfactory recognition, emerging during early stages of the disease and worsening while the disease progresses. The social transmission of food preferences (STFP) task is based on a rudimentary form of communication between rodents concerning distant foods dependent on the transmission of olfactory cues. Healthy wild-type mice would prefer to eat a novel, flavored food that was previously cued by a conspecific, and this food preference would be hampered in transgenic AD mice, such as the APP/PS1 model. Indeed, a strong preference for the cued food in C57Bl6/J mice of 3 months of age was found, and this was reduced in 3 months old transgenic APP/PS1 mice. In summary, STFP task could be a powerful measure to be integrated in present subclinical detection assays of AD.

This paper presents a protocol for the investigation of social transmission of food preference in mice. The advantages and possible applications for this procedure, for instance, in detecting early changes in AD mouse models, are highlighted. To conclude, interpretation of the results in light of critical details are discussed.

Olfactory recognition deficits are suggested to be able to serve as clinical marker to differentiate Alzheimer&#39;s disease (AD) subjects from healthy ...

Control of Eating Behavior Using a Novel Feedback System

Subjects eat food from a plate that sits on a scale connected to a computer that records the weight loss of the plate during the meal and makes up a curve of food intake, meal duration and rate of eating modeled by a quadratic equation. The purpose of the method is to change eating behavior by providing visual feedback on the computer screen that the subject can adapt to because her/his own rate of eating appears on the screen during the meal. The data generated by the method is automatically analyzed and fitted to the quadratic equation using a custom made algorithm. The method has the advantage of recording eating behavior objectively and offers the possibility of changing eating behavior both in experiments and in clinical practice. A limitation may be that experimental subjects are affected by the method. The same limitation may be an advantage in clinical practice, as eating behavior is more easily stabilized by the method. A treatment that uses this method has normalized body weight and restored the health of several hundred patients with anorexia nervosa and other eating disorders and has reduced the weight and improved the health of severely overweight patients.

Subjects eat food from a plate placed on a scale connected to a computer that records the weight loss of the plate during the meal. Feedback on the computer screen allows the subject to adapt her/his eating behavior to reference curves thus normalizing body weight.

Subjects eat food from a plate that sits on a scale connected to a computer that records the weight loss of the plate during the meal and makes up a curve ...

Concept Development and Use of an Automated Food Intake and Eating Behavior Assessment Method

The vast majority of dietary and eating behavior assessment methods are based on self-reports. They are burdensome and also prone to measurement errors. Recent technological innovations allow for the development of more accurate and precise dietary and eating behavior assessment tools that require less effort for both the user and the researcher. Therefore, a new sensor-based device to assess food intake and eating behavior was developed. The device is a regular dining tray equipped with a video camera and three separate built-in weighing stations. The weighing stations measure the weight of the bowl, plate, and drinking cup continuously over the course of a meal. The video camera positioned to the face records eating behavior characteristics (chews, bites), which are analyzed using artificial intelligence (AI)-based automatic facial expression software. The tray weight and the video data are transported at real-time to a personal computer (PC) using a wireless receiver. The outcomes of interest, such as the amount eaten, eating rate and bite size, can be calculated by subtracting the data of these measures at the timepoints of interest. The information obtained by the current version of the tray can be used for research purposes, an upgraded version of the device would also facilitate the provision of more personalized advice on dietary intake and eating behavior. Contrary to the conventional dietary assessment methods, this dietary assessment device measures food intake directly within a meal and is not dependent on memory or the portion size estimation. Ultimately, this device is therefore suited for daily main meal food intake and eating behavior measures. In the future, this technology based dietary assessment method can be linked to health applications or smart watches to obtain a complete overview of exercise, energy intake, and eating behavior.

This protocol shows and explains a new technology-based dietary assessment method. The method consists of a dining tray with multiple built-in weighing scales and a video camera. The device is unique in the sense that it incorporates automated measures of food and drink intake and eating behavior over the course of a meal.

The vast majority of dietary and eating behavior assessment methods are based on self-reports. They are burdensome and also prone to measurement errors. ...

Oral Biofilm Sampling for Microbiome Analysis in Healthy Children

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition leads to a better understanding of cariogenic and periopathogenic mechanisms. Microbial changes taking place in the oral cavity during childhood are of interest for several reasons. The evolution of the child oral microbiota and shifts in its composition need to be analyzed further to understand and possibly prevent the onset of disease. At the same time, advanced knowledge of the natural composition of oral biofilm is needed. Early stages of caries-free permanent dentition with healthy gums provide a widely unaffected subgingival habitat that can serve as an in situ baseline for studying features of oral health and disease. Analysis of children's oral biofilm during different stages in life is thus an important theme in the field. Modern molecular analysis methods can provide comprehensive information about the bacterial diversity of such biofilms. To enable microbiota data comparison, it is important to standardize each step in the procedure for molecular data generation. This procedure spans from clinical sampling, Next Generation Sequencing (NGS), bioinformatic data processing, to taxonomic interpretation. One of the most critical factors here is biofilm sampling. Sampling in children is even more challenging in particular due to limited space in subgingival areas. We thus focus on the use of paper points for subgingival sampling. This article provides a detailed protocol for oral biofilm sampling of the subgingival sulcus, the mucosa, and saliva in children.

Changes in the oral microbiome throughout childhood are of growing interest. Comparison of different microbiome studies reveals a lack of standardized sampling protocols. Limited space makes sampling the sound subgingival sulcus of children challenging. Paper point sampling is presented here in detail as the method of choice for this area.

Oral biofilm and its molecular analysis provide a basis for investigating various dental research and clinical questions. Knowledge of biofilm composition ...

Medicine

Palatable Western-style Cafeteria Diet as a Reliable Method for Modeling Diet-induced Obesity in Rodents

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health burden of obesity and its comorbid conditions highlight the need for better understanding of its pathogenesis, yet ethical constraints limit studies in humans. To this end externally valid models of obesity in laboratory animals are essential for the understanding of being overweight and obesity.&#160;While many species have been used to model the range of changes that accompany obesity in humans, rodents are most commonly used. Our laboratory has developed a western-style cafeteria diet that consistently leads to considerable weight gain and markers of metabolic disease in rodents. The diet exposes rodents to a variety of highly palatable foods to induce hyperphagia, modeling the modern western food environment. This diet rapidly induces weight gain and body fat accumulation in rats allowing for the study of effects of overeating and obesity. While the cafeteria diet may not provide the same control over macronutrient and micronutrient profile as purified high-fat or high-fat, high-sugar diets, the cafeteria diet typically induces a more severe metabolic phenotype than that observed with purified diets and is more in line with metabolic disturbances observed in the overweight and obese human population.

This protocol describes the use of a highly palatable, western-style cafeteria diet to model overeating and obesity in rodents. Here, we provide a detailed outline of food selection, preparation and measurement, and explain methodological factors that assist in generating a robust and reproducible phenotype.

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health ...

Sucrose Preference and Novelty-Induced Hypophagia Tests in Rats using an Automated Food Intake Monitoring System

The prevalence and incidence of depressive disorders are rising worldwide, affecting about 322 million individuals, underlining the need for behavioral studies in animal models. In this protocol, to study depression-like and anhedonic behavior in rats, the established sucrose preference and novelty-induced hypophagia tests are combined with an automated food and liquid intake monitoring system. Prior to testing, in the sucrose preference paradigm, male rats are trained for at least 2 days to consume a sucrose solution in addition to tap water. During the test, rats are again exposed to water and sucrose solution. Consumption is registered every second by the automated system. The ratio of sucrose to total water intake (sucrose preference ratio) is a surrogate parameter for anhedonia. In the novelty-induced hypophagia test, male rats undergo a training period in which they are exposed to a palatable snack. During training, rodents show a stable baseline snack intake. On test day, the animals are transferred from home cages into a fresh, empty cage representing a novel unknown environment with access to the known palatable snack. The automated system records the total intake and its underlying microstructure (e.g., latency to approaching the snack), providing insight into anhedonic and anxious behaviors. The combination of these paradigms with an automated measuring system provides more detailed information, along with higher accuracy by reducing measuring errors. However, the tests use surrogate parameters and only depict depression and anhedonia in an indirect manner.

Presented here is a protocol to study depression-like and anhedonic behavior in rats. It combines two well-established behavioral methods, the sucrose preference and novelty-induced hypophagia tests, with an automated food and liquid intake monitoring system, to indirectly investigate rodent behavior using surrogate parameters.

The prevalence and incidence of depressive disorders are rising worldwide, affecting about 322 million individuals, underlining the need for behavioral ...

Assessing Strength Exercise Volume and Its Impact on Insulin Sensitivity in Obesity

Randomized Controlled Trial to Study the Acute Effects of Strength Exercise on Insulin Sensitivity in Obese Adults

An acute session of strength exercise (SE) ameliorates insulin sensitivity (IS) for several hours; however, the effects of SE volume (i.e., number of sets) have not been studied thoroughly. Although it is intuitive that some SE is better than none, and more is better than some for the improvement of IS, high-volume sessions might be challenging for diseased populations to complete, especially obese adults, for whom even a brisk walk can be challenging. This protocol details a randomized clinical trial to assess the acute effects of SE on IS in obese adults. The inclusion criteria are body mass index &#62;30 kg/m2, central obesity (waist circumference &#62;88 cm and &#62;102 cm for women and men, respectively), and age &#62;40 years. Participants will be familiarized with the SE (7 exercises targeting major muscle groups) and then will perform three sessions in a randomized order: session 1 - high-volume session (3 sets/exercise); session 2 - low-volume session (1 set/exercise); session 3 - control session (no exercise). Diet will be controlled the day before and on the day of the sessions. Sessions will be completed at night, and an oral glucose tolerance test will be performed the next morning, from which several indexes of IS will be derived, such as the area under the curve (AUC) of glucose and insulin, the Matsuda index, the Cederholm index, the muscle IS index, and the Gutt index. Based on pilot studies, we expect ~15% improvement in IS (insulin AUC, and Matsuda and Cederholm indexes) after the high-volume session, and ~8% improvement after the low-volume session compared to the control session. This study will benefit individuals who find high-volume SE sessions challenging but still aim to improve their IS by investing 1/3 of their time and effort.

Author Spotlight: Exploring the Impact of Reduced Resistance Exercise Volume on Metabolic Health 

This study describes a randomized controlled trial protocol aiming at assessing the acute effects of strength exercise volume on insulin sensitivity in obese individuals.

An acute session of strength exercise (SE) ameliorates insulin sensitivity (IS) for several hours; however, the effects of SE volume (i.e., number of ...

The Caco-2 Cell Bioassay for Measurement of Food Iron Bioavailability

Knowledge of Fe bioavailability is critical to the assessment of the nutritional quality of Fe in foods. In vivo measurement of Fe bioavailability is limited by cost, throughput, and the caveats inherent to isotopic labeling of the food Fe. Thus, there exists a critical need for an approach that is high-throughput and cost-effective. The Caco-2 cell bioassay was developed to satisfy this need. The Caco-2 cell bioassay for Fe bioavailability utilizes simulated gastric and intestinal digestion coupled with culture of a human intestinal epithelial cell line known as Caco-2. In Caco-2 cells, Fe uptake stimulates the intracellular formation of ferritin, an Fe storage protein easily measured by enzyme-linked immunosorbent assay (ELISA). Ferritin forms in proportion to Fe uptake; thus, by measuring Caco-2 cell ferritin production, one can assess intestinal Fe uptake from simulated food digests into the enterocyte. 
Via this approach, the model replicates the key initial step that determines food Fe bioavailability. Since its inception in 1998, this model approach has been rigorously compared to factors known to influence human Fe bioavailability. Moreover, it has been applied in parallel studies, with three human efficacy studies evaluating Fe biofortified crops. In all cases, the bioassay correctly predicted the relative amounts of Fe bioavailability from the factors, crops, and overall diet. This paper provides detailed methods on Caco-2 cell culture coupled with the in vitro digestion process and cell ferritin ELISA necessary to conduct the Caco-2 cell bioassay for Fe bioavailability.

The Caco-2 cell bioassay for iron (Fe) bioavailability represents a cost-effective and versatile approach to assess Fe bioavailability from foods, food products, supplements, meals, and even diet regimens. Thoroughly validated to human studies, it represents the state of the art for studies of Fe bioavailability.

Knowledge of Fe bioavailability is critical to the assessment of the nutritional quality of Fe in foods. In vivo measurement of Fe bioavailability is ...