Perspectives of nonmajor undergraduate students on the impact of group discussions in learning physiology.

A 3-year study (2017-2019) was conducted to obtain the views of nonmajor undergraduate students about discussions in learning physiology. The teaching methods used were lecture only (lecture), group discussion alone (discussion), and a combination of lecture and discussion (lecture + discussion). Students were assigned homework in a textbook, and they did not have access to textbook/notes during discussions. Under these conditions, 58% of students indicated that they learned best with lecture + discussion strategy, compared with 49% for lecture and 18% for discussion approaches. Remarkably, 61% of students said the discussion did not enhance learning; by comparison, 35% and 14% had the same views about lecture and lecture + discussion, respectively. Furthermore, if given the opportunity to choose a teaching/learning environment, 66% of students would select lecture + discussion, 33% would pick lecture, and only 6% would choose discussion setting. As many as 77% of students would reject the discussion setting if given the choice. The opinions of students were similar irrespective of their expected grades (whether A, B, or C); however, greater proportions of B or C students disliked discussion than A students. Thus, whereas 63% of A students disliked discussion, 81% of B students and 83% of C students disliked it. Also, 64% of students indicated that they would have been poorly prepared for classes without assigned homework. Essential outcomes of this study include undergraduates viewed the lecture + discussion setting as a supportive/desirable environment for learning physiology, and they consistently rated the lecture method higher than the discussion-only approach. Students did not relish learning physiology in a discussion-only setting. These findings may help in establishing teaching/learning environments from the student's perspective.NEW & NOTEWORTHY This article reports perspectives of nonmajor undergraduates about group discussions in learning physiology. Three teaching methods were used: traditional lecture alone (lecture), discussion alone (discussion), and combined lecture and discussion (lecture + discussion). Students rated lecture + discussion setting as the most conducive for learning. The rank order of student preference for learning environment was, first, lecture + discussion; second, lecture; and third, discussion. These opinions were similar irrespective of expected grades in the course. Enjoyment of the teaching/learning process and environment is important to students.

Impact of combination of short lecture and group discussion on the learning of physiology by nonmajor undergraduates.

This study assessed the impact of an "active learning" strategy employed alone or in combination with traditional lectures on the learning of mammalian physiology by undergraduate students. The study investigated the impact of three teaching strategies, namely 1) traditional lecture, 2) group discussion alone, and 3) combination of lecture and group discussion. For all strategies, students were given homework in a textbook and they completed written assignments before each session. Every student led the discussion of at least one assigned theme during each group session. The students had no access to the textbook or notes during group discussions. Four examinations (3 in-semester and a final) assessed the students' knowledge of fundamental concepts of physiology of specific organ systems. Part of the final examination reassessed knowledge of previously tested topics. The results show that the teaching modality employed to introduce physiology topics influenced students' learning. The average marginal effect of the lecture + discussion modality (average improvement linked to lecture + discussion strategy) on students' performance was 6.45% [95% confidence interval (CI95) (4.73, 8.16), P = 1.74 × 10-13], and the average improvement associated with the discussion-only modality was 5.5% [CI95 (3.84, 7.16), P = 7.84 × 10-11]. On average, all class ranks performed better on materials covered under active learning settings than under lecture-only conditions. Moreover, students' performance under combined lecture and discussion conditions is predictive of their overall performance in the course. The results support the positive effect of student-centered learning and demonstrate the efficacy of a combination of lectures and group discussions on learning of physiology by nonmajor students.NEW & NOTEWORTHY The purpose of this study was to evaluate the effect of group discussion on the learning of mammalian physiology by nonmajor undergraduate students. Combining traditional lectures with group discussions increased the active participation of students in class and improved their learning of physiology, as measured by the results of in-semester and final examinations. The active learning technique benefited all class ranks on average.

Factors necessary to produce basoapical polarity in human glandular epithelium formed in conventional and high-throughput three-dimensional culture: example of the breast epithelium.

BACKGROUND: Basoapical polarity in epithelia is critical for proper tissue function, and control of proliferation and survival. Cell culture models that recapitulate epithelial tissue architecture are invaluable to unravel developmental and disease mechanisms. Although factors important for the establishment of basal polarity have been identified, requirements for the formation of apical polarity in three-dimensional tissue structures have not been thoroughly investigated. RESULTS: We demonstrate that the human mammary epithelial cell line-3522 S1, provides a resilient model for studying the formation of basoapical polarity in glandular structures. Testing three-dimensional culture systems that differ in composition and origin of substrata reveals that apical polarity is more sensitive to culture conditions than basal polarity. Using a new high-throughput culture method that produces basoapical polarity in glandular structures without a gel coat, we show that basal polarity-mediated signaling and collagen IV are both necessary for the development of apical polarity. CONCLUSION: These results provide new insights into the role of the basement membrane, and especially collagen IV, in the development of the apical pole, a critical element of the architecture of glandular epithelia. Also, the high-throughput culture method developed in this study should open new avenues for high-content screening of agents that act on mammary tissue homeostasis and thus, on architectural changes involved in cancer development.

Phytic acid increases mucin and endogenous amino acid losses from the gastrointestinal tract of chickens.

The influence of the form of phytic acid on the regulation of mucin and endogenous losses of amino acids, nitrogen and energy in chickens was investigated. Forty-eight 10-week-old male broilers were grouped by weight into eight blocks of six cages with one bird per cage. Birds received by intubation six dextrose-based combinations of phytic acid and phytase arranged in a 3 x 2 factorial consisting of phytic acid form (no phytic acid, 1.0 g free phytic acid or 1.3 g magnesium-potassium phytate) and phytase (0 or 1000 units). Each bird received the assigned combination added to 25 g dextrose at each of the two feedings on the first day of experimentation. All excreta were collected continuously for 54 h following feeding and frozen until analysed. Frozen excreta were thawed, pooled for each bird, lyophilised, ground, and analysed for DM, energy, nitrogen, amino acids, mucin, and sialic and uric acids. Chickens fed either magnesium-potassium phytate or free phytic acid showed increased (P < 0.05) loss of crude mucin and sialic acid. The amount of crude mucin lost was significantly greater (P < 0.05) with magnesium-potassium phytate than with free phytic acid treatment. Both phytic acid treatments also increased (P < 0.05) endogenous loss of threonine, proline and serine. In conclusion, the form of phytic acid fed to chickens affects the extent of mucin and endogenous amino acid losses from the gastrointestinal tract.

Quadratic form: a robust metric for quantitative comparison of flow cytometric histograms.

Comparison of fluorescence distributions is a fundamental part of the analysis of flow cytometric data. This approach is applied to detect differences between control and test sample and thus analyze a biological response. Comparison of standard test samples over time provides an estimate of instrument stability for quality control. However, application of statistical methods of distribution comparison in flow cytometry is difficult owing to instrument noise and the complex shape of intensity distributions. We applied quadratic form (QF) as a mathematical metric for comparison of flow cytometry histograms. QF operates on histograms as vectors and calculates the total distance in an interbin manner using a matrix of distances between single histogram bins. Euclidean interbin distance and histograms normalized to unity were used. Critical values corresponding to 95% significance level were calculated using Monte-Carlo simulation and single-maximum Gaussian distributions populated with several numbers of events. The QF statistic was then validated for non-Gaussian single-maximum distributions and multiple-maxima distributions. We determined that the critical values for Gaussian distributions depended on standard deviations and number of events in the compared histograms. A simple empirical function was constructed to characterize this dependence. Furthermore, it was verified that critical values (corresponding to 95% significance) for non-Gaussian histograms were similar to values for the Gaussian histograms characterized by the same standard deviation. We applied the QF statistic to estimate the differences between histograms of DNA content (ploidy) in cells of old and young leaf tissue of Brassica campestris. Furthermore, we quantified differences in fluorescence intensity in immunostaining of human lymphocytes. Quadratic form (QF) provides a true (mathematical) metric for estimation of distance between flow cytometry histograms of arbitrary shape. QF can be applied as a statistical test for estimation of significance of the distance measure. The respective critical values depend only on the number of events and standard deviations of compared histograms and are not affected by distribution shape. Therefore, applications of QF do not require assumptions concerning distribution shape and can be easily implemented in practice. This notion was confirmed using empirical distributions of DNA content in plant tissue and distributions of immunofluorescence in human cells.

Phytates reduce uptake of leucine and glutamate but not lysine and glucose from the intestinal lumen of chickens: short communication.

An investigation into the influence of phytates on the in situ absorption of amino acids (lysine, glutamate and leucine) and glucose from the intestinal lumen of 3-week-old chickens was carried out. Birds were anaesthetised and the intestines exteriorised. Uptake of 5 mM of each nutrient over a 4-min period was measured in the presence of four phytate concentrations (0, 50, 250 and 500 mM). Five birds were used for each nutrient at each concentration of phytate tested. Leucine uptake decreased linearly (P < 0.001) and that of glutamate showed a tendency to decrease (P = 0.055) as the phytate concentration increased. Absorption of lysine and glucose were unaffected by the presence of phytate. In conclusion, phytate in the small intestinal lumen exerted a depressive effect on the absorption of specific free amino acids from the lumen. Its depressive effect was greatest for leucine followed by glutamate, and phytate had little effect on the absorption of lysine.

Compression of fluorescence microscopy images based on the signal-to-noise estimation.

Modern microscopic techniques like high-content screening (HCS), high-throughput screening, 4D imaging, and multispectral imaging may involve collection of thousands of images per experiment. Efficient image-compression techniques are indispensable to manage these vast amounts of data. This goal is frequently achieved using lossy compression algorithms such as JPEG and JPEG2000. However, these algorithms are optimized to preserve visual quality but not necessarily the integrity of the scientific data, which are often analyzed in an automated manner. Here, we propose three observer-independent compression algorithms, designed to preserve information contained in the images. These algorithms were constructed using signal-to-noise ratio (SNR) computed from a single image as a quality measure to establish which image components may be discarded. The compression efficiency was measured as a function of image brightness and SNR. The alterations introduced by compression in biological images were estimated using brightness histograms (earth's mover distance (EMD) algorithm) and textures (Haralick parameters). Furthermore, a microscope test pattern was used to assess the effect of compression on the effective resolution of microscope images.

Loss of image quality in photobleaching during microscopic imaging of fluorescent probes bound to chromatin.

Prolonged excitation of fluorescent probes leads eventually to loss of their capacity to emit light. A decrease in the number of detected photons reduces subsequently the resolving power of a fluorescence microscope. Adverse effects of fluorescence intensity loss on the quality of microscopic images of biological specimens have been recognized, but not determined quantitatively. We propose three human-independent methods of quality determination. These techniques require no reference images and are based on calculation of the actual resolution distance, information entropy, and signal-to-noise ratio (SNR). We apply the three measures to study the effect of photobleaching in cell nuclei stained with propidium iodide (PI) and chromomycin A3 (CA3) and imaged with fluorescence confocal microscopy. We conclude that the relative loss of image quality is smaller than the corresponding decrease in fluorescence intensity. Furthermore, the extent of quality loss is related to the optical properties of the imaging system and the noise characteristics of the detector. We discuss the importance of these findings for optimal registration and compression of biological images.

Confocal fluorescence imaging of photosensitized DNA denaturation in cell nuclei.

The double-stranded helical structure of DNA is maintained in part by hydrogen bonds between strands and by stacking interactions between adjacent purine and pyrimidine bases in one strand. The transition (denaturation) from a double-stranded (ds) to a single-stranded (ss) form can be induced in isolated DNA or fixed cells by exposure to elevated temperatures, alkali or acids, aprotic or nonpolar solvents or some drugs. We report here that DNA denaturation can occur in situ in cell nuclei as a result of interaction between light and an intercalated dye, acridine orange or ethidium bromide. This DNA photodenaturation was probed using metachromatic properties of acridine orange and imaged by fluorescence confocal microscopy. Furthermore, an empirical kinetic model was developed to separate changes of acridine orange luminescence intensities caused by photobleaching from those that were a result of DNA denaturation. We investigated the influence of oxygen on these phenomena and propose a mechanism by which photodenaturation may occur.

Three-dimensional cell culture to model epithelia in the female reproductive system.

In vitro 3-dimensional (3D) cell cultures produce valuable models that mimic 3D tissue organization and function and enhance the understanding of cell/tissue function under normal and pathological situations. Tissue function depends on the interactions between cells and the extracellular matrix; thus, effective 3D cell cultures rely on the use of appropriate extracellular matrix cues. Noticeable progress in 3D cell culture was obtained from studies with epithelial cells from organs of the female reproductive system including the mammary glands, the uterus, and the ovaries. These models show that replicating normal tissue organization in the resting phase is a prerequisite for appropriate physiological and pathological investigations. The authors' goals are to explain the importance of mimicking detailed aspects of normal epithelial organization and function, such as basoapical polarity, in 3D cell culture and to discuss how effective 3D cell culture models can lead to meaningful applications in reproductive biology.

Precision of light intensity measurement in biological optical microscopy.

Standardization and calibration of optical microscopy systems have become an important issue owing to the increasing role of biological imaging in high-content screening technology. The proper interpretation of data from high-content screening imaging experiments requires detailed information about the capabilities of the systems, including their available dynamic range, sensitivity and noise. Currently available techniques for calibration and standardization of digital microscopes commonly used in cell biology laboratories provide an estimation of stability and measurement precision (noise) of an imaging system at a single level of signal intensity. In addition, only the total noise level, not its characteristics (spectrum), is measured. We propose a novel technique for estimation of temporal variability of signal and noise in microscopic imaging. The method requires registration of a time series of images of any stationary biological specimen. The subsequent analysis involves a multi-step process, which separates monotonic, periodic and random components of every pixel intensity change in time. The technique allows simultaneous determination of dark, photonic and multiplicative components of noise present in biological measurements. Consequently, a respective confidence interval (noise level) is obtained for each level of signal. The technique is validated using test sets of biological images with known signal and noise characteristics. The method is also applied to assess uncertainty of measurement obtained with two CCD cameras in a wide-field microscope.

Basal lamina of ovarian follicle regulates an inward Cl(-) current in differentiated granulosa cells.

Patch-clamp experiments were conducted to study the effects of basal lamina (basement membrane) of preovulatory chicken ovarian follicle on membrane currents in differentiated chicken granulosa cells in a homologous system. The membrane capacitance (measure of total membrane area) was smaller in cells cultured on intact basal lamina than that of control cells. The granulosa cells expressed outward and two inward currents. A small fraction of the cells (3%) expressed only a transient fast-activating and -inactivating inward current carried by Ca(2+). The majority of the cells, however, expressed a slowly activating and inactivating inward current (carried by Cl(-)) that was superimposed on the transient Ca(2+) current. All cells expressed an outward current characteristic of the delayed-rectifier K(+) current. The removal of extracellular Ca(2+) led to elimination of the slow inward Cl(-) current, indicating that it is a Ca(2+)-dependent Cl(-) current. Both peak amplitude and current density of the inward Cl(-) current were significantly lower in cells cultured on freshly isolated intact basal lamina (or basal lamina stored at 4 degrees C for 12 mo) than those of control cells; however, basal lamina had no significant effect on the density of the outward current. Similar to the observations made for intact basal lamina, solubilized basal lamina suppressed the inward Cl(-) current in differentiated granulosa cells. These data show that homologous basal lamina modulates a Ca(2+)-dependent Cl(-) current in differentiated granulosa cells. These findings provide a partial explanation for the mechanisms that subserve the reported effects of basal lamina (basement membrane) on the metabolic functions of differentiated granulosa cells.

Effect of basal lamina of ovarian follicle on T- and L-type Ca(2+) currents in differentiated granulosa cells.

Patch clamp experiments were conducted to study the effects of basal lamina (basement membrane) of chicken ovarian follicle on membrane Ca(2+) currents in differentiated chicken granulosa cells in a homologous system. The whole cell patch clamp technique was used to simultaneously monitor membrane capacitance (an indirect measure of total cell surface area) and currents flowing through voltage-dependent Ca(2+) channels (using Ba(2+) as the charge carrier). Membrane capacitance was smaller in cells incubated on intact basal lamina than in control cells (incubated on tissue culture-treated plastic substratum). Granulosa cells expressed both T- and L-type Ca(2+) currents, and the amplitudes of the currents in cells incubated on intact basal lamina were significantly lower than those of control cells. Also, granulosa cells incubated on intact basal lamina were found to have significantly lower T- or L-type Ca(2+) current densities than control cells. Intact basal lamina that had been stored for 12 mo produced effects on T- and L-type Ca(2+) currents similar to those caused by freshly isolated basal lamina. The basal lamina was solubilized completely in one step and used to coat glass coverslips (uncoated glass coverslips served as controls). Granulosa cells incubated on coverslips precoated with solubilized basal lamina assumed spherical shape similar to those incubated on intact basal lamina. Similar to the observations made for intact basal lamina, the solubilized basal lamina suppressed T- and L-type Ca(2+) currents in the differentiated granulosa cells. Moreover, fibronectin, laminin, and type IV collagen, obtained from commercial sources, attenuated T- and L-type Ca(2+) currents in the differentiated granulosa cells. This interplay between basal lamina and Ca(2+) currents may be one mechanism that subserves the effects of the matrix material on metabolic functions of granulosa cells.

Postnatal ontogeny of kinetics of porcine jejunal brush border membrane-bound alkaline phosphatase, aminopeptidase N and sucrase activities.

Our objectives were to determine postnatal changes in the maximal enzyme activity (V(max)) and enzyme affinity (K(m)) of jejunal mucosal membrane-bound alkaline phosphatase, aminopeptidase N and sucrase using a porcine model which may more closely resemble the human intestine. Jejunal brush border membrane was prepared by Mg(2+)-precipitation and differential centrifugation from pigs of suckling (8 days), weaning (28 days), post-weaning (35 days) and adult (70 days) stages. p-Nitrophenyl phosphate (0-8 mM), L-alanine-p-nitroanilide hydrochloride (0-28 mM) and sucrose (0-100 mM) were used in alkaline phosphatase, aminopeptidase N and sucrase kinetic measurements. V(max) of alkaline phosphatase was the lowest in the adult (4.27 micromol.mg(-1) protein.min(-1)), intermediate in the suckling (9.75 micromol.mg(-l) protein.min(-l)) and the highest in the weaning and post-weaning stage (12.83 and 10.40 micromol.mg(-l) protein.min(-l)). K(m) of alkaline phosphatase was high in the suckling and weaning stages (5.14 and 9.93 mM) and low in the adult (0.66 mM). V(max) of aminopeptidase N was low in the suckling (7.04 micromol.mg protein(-1).min(-1)) and high in the post-weaning stage (13.36 micromol.mg(-l) protein.min(-l)). K(m) of aminopeptidase N was the highest in the two weaning stages (2.96 and 3.39 mM), intermediate in the adult (2.33 mM) and the lowest in the suckling stage (1.66 mM). V(max) of sucrase increased from the suckling to the adult (0.48-1.30 micromol.mg(-l) protein.min(-l)). K(m) of sucrase ranged from 11.19 to 16.57 mM. There are dramatic postnatal developmental changes in both the maximal enzyme activity and enzyme affinity of jejunal brush border membrane-bound alkaline phosphatase, aminopeptidase N and sucrase in the pig.