John A. McNulty

Serotoninergic and melatoninergic systems are fully expressed in human skin

We investigated the cutaneous expression of genes and enzymes responsible for the multistep conversion of tryptophan to serotonin and further to melatonin. Samples tested were human skin, normal and pathologic (basal cell carcinoma and melanoma), cultured normal epidermal and follicular melanocytes, melanoma cell lines, normal neonatal and adult epidermal and follicular keratinocytes, squamous cell carcinoma cells, and fibroblasts from dermis and follicular papilla. The majority of the samples showed simultaneous expression of the genes for tryptophan hydroxylase, arylalkylamine N‐acetyltransferase (AANAT), and hydroxyindole‐Omethyltransferase (HIOMT). The products of AANAT activity were identified by RP‐HPLC with fluorimetric detection in human skin and in cultured normal and malignant melanocytes and immortalized keratinocytes; HIOMT activity was detected in human skin, keratinocytes, and melanoma cells. N‐acetylserotonin (NAS) was detected by RP‐HPLC in human skin extracts. NAS identity was confirmed further by LC/MS in keratinocytes. In conclusion, we provide evidence that the human skin expresses intrinsic serotonin and melatonin biosynthetic pathways.

Evaluation of computer-aided instruction in a gross anatomy course: a six-year study.

Web‐based computer‐aided instruction (CAI) has become increasingly important to medical curricula. This multi‐year study investigated the effectiveness of CAI and the factors affecting level of individual use. Three CAI were tested that differed in specificity of applicability to the curriculum and in the level of student interaction with the CAI. Student personality preferences and learning styles were measured using the Meyers Briggs Type Indicator (MBTI) and Kolbs Learning Style Inventory (LSI). Information on “computer literacy” and use of CAI was collected from student surveys. Server logs were used to quantify individual use of respective CAI. There was considerable variability in the level of utilization of each CAI by individual students. Individual use of each CAI differed and was associated with gender, MBTI preferences and learning style, but not with “computer literacy.” The majority of students found the CAI useful for learning and used the CAI by themselves. Students who accessed the CAI resources most frequently scored significantly higher on exams compared with students who never accessed the resources. Our results show that medical students do not uniformly use CAI developed for their curriculum and this variability is associated with various attributes of individual students. Our data also provide evidence of the importance of understanding student preferences and learning styles when implementing CAI into the curriculum. Anat Sci Ed 2:2–8, 2009.

Immunocytochemical and electron-microscopic characterization of macrophage/microglia cells and expression of class II major histocompatibility complex in the pineal gland of the rat

Interstitial cells in the pineal gland of the rat were characterized immunocytochemically using the monoclonal antibodies MRC OX-42 and ED1 for macrophages/microglia, and MRC OX-6, which recognizes major histocompatibility complex (MHC) class II antigen. A polyclonal antibody against GFAP was used to identify astrocytes. Cells immunopositive for OX-42 and/or ED1 were distributed throughout the gland; they extended processes primarily along the perivascular spaces and occasionally within the parenchyma of the gland. Ultrastructurally, these OX-42-positive cells were characterized by a nucleus with sparse heterochromatin and cytoplasmic vacuoles/lysosomes. Cells expressing MHC class II antigen had a distribution and morphology similar to OX-42-immunopositive cells, suggesting that pineal macrophages/microglia play a role as antigen-presenting cells. GFAP-positive astrocytes were concentrated at the proximal end of the pineal where the pineal stalk enters the gland. The occurrence of antigenpresenting cells in the circumventricular neuroendocrine gland has important functional implications as these cells may be mediators of neuroimmunomodulatory mechanisms, and involved in certain disease states such as autoimmune pinealitis.

An analysis of lecture video utilization in undergraduate medical education: associations with performance in the courses.

BackgroundIncreasing numbers of medical schools are providing videos of lectures to their students. This study sought to analyze utilization of lecture videos by medical students in their basic science courses and to determine if student utilization was associated with performance on exams.MethodsStreaming videos of lectures (n = 149) to first year and second year medical students (n = 284) were made available through a password-protected server. Server logs were analyzed over a 10-week period for both classes. For each lecture, the logs recorded time and location from which students accessed the file. A survey was administered at the end of the courses to obtain additional information about student use of the videos.ResultsThere was a wide disparity in the level of use of lecture videos by medical students with the majority of students accessing the lecture videos sparingly (60% of the students viewed less than 10% of the available videos. The anonymous student survey revealed that students tended to view the videos by themselves from home during weekends and prior to exams. Students who accessed lecture videos more frequently had significantly (p < 0.002) lower exam scores.ConclusionWe conclude that videos of lectures are used by relatively few medical students and that individual use of videos is associated with the degree to which students are having difficulty with the subject matter.

Synaptic ribbons in the pineal organ of the goldfish: Circadian rhythmicity and the effects of constant light and constant darkness

SummarySynaptic ribbons in photoreceptor cells of the goldfish pineal organ undergo significant daily changes in their length, distance from the plasma membrane, and number per unit area of pineal end-vesicle. The rhythms persist in fish exposed to constant darkness. Constant light abolishes the rhythms in length and distance of synaptic ribbons from the plasmalemma, but has little effect on numerical changes over a 24-h cycle. These findings suggest that synaptic ribbons in the pineal organ of lower vertebrates might be useful as indicators of metabolic activity.

Enriched immune-environment of blood-brain barrier deficient areas of normal adult rats.

The circumventricular organs (CVOs) in the brain are without a blood-brain barrier (BBB) and as such directly exposed to blood plasma constituents and blood-borne pathogens. In light of previous studies showing discrepancies regarding the immunocompetence of these organs, we initiated the present study to provide a comprehensive immunohistochemical analysis of the cellular expression of immune-associated antigens within the pineal gland, area postrema and the subfornical organ. In all CVOs, subpopulations of cells morphologically similar to complement receptor type 3 immunoreactive microglial/macrophage cells expressed major histocompatibility complex (MHC) class II antigen, leucocyte common antigen (LCA/CD45), as well as CD4 and ED1 antigen. Based on morphological criteria the MHC class II antigen expressing cells could be grouped into a major population of classical parenchymal and perivascular ramified microglial cells and a minor population presenting itself as scattered or small groups of rounded macrophage-like cells. CD4 and ED1 antigen were expressed by both cell types. CD45 was preferentially expressed by macrophage-like cells. MHC class I antigen was expressed by the vascular endothelium in both BBB-protected and BBB-deficient areas and was additionally present as a lattice-like network throughout the BBB-deficient parenchyma in all CVOs. The results suggest that the BBB-free areas of the brain besides being constantly surveyed by blood-borne macrophages, possess an intrinsic immune surveillance system based on resting and activated microglial cells, which may function as a non-endothelial, cellular barrier against blood-borne pathogens.

Effect of suprachiasmatic ablation on the daily temperature rhythm.

Abstract To evaluate the role of the suprachiasmatic nuclei (SCN) in maintenance of body temperature, the rectal temperature of 5 SCN ablated and 8 intact female rats were measured every 4 h over a 48-h period (80 days postsurgery). Intact as well as SCN ablated rats showed significant 24-h fluctuations in body temperature. SCN ablation altered neither the phase nor the level (24-h mean) of the rhythm.

Circadian analysis of mononuclear cells in the rat following pinealectomy and superior cervical ganglionectomy

The role of the pineal gland and adrenocorticosteroids in circadian rhythmicity of immune function was investigated in Sprague-Dawley rats that either had been pinealectomized (PX) or had undergone superior cervical ganglionectomy (SCGX), which functionally denervates the pineal. For both experiments, conducted between August and October, rats were entrained to a 10-h light:14-h dark cycle and fed ad libitum. The circadian rhythm in monocytes (ANOVA, p less than .01 for Control + Sham + PX samples) tended to parallel that of plasma corticosteroid levels (ANOVA, p less than .01) with the peak during early dark and the nadir at early light. In comparison, suppressor/cytotoxic T cells and B cells tended to be more frequent during early light with minimum levels during early dark (ANOVA, p less than .05 and .01, respectively, for Control + Sham + PX samples). Natural killer (NK) activity in control animals varied significantly (ANOVA, p less than .05) over the light:dark cycle with peak activity during early dark in the SCGX experiment, but was only slightly elevated during late dark in the PX experiment. PX animals exhibited increased frequency of NK cells, but only in samples collected during the day (t test, p less than .05). This effect was not observed in sham-operated PX controls and in SCGX animals. Neither PX nor SCGX had a significant (ANOVA) effect on plasma corticosterone levels. These observations in the rat are consistent with findings in mice and humans that corticosteroids play an important general role in circadian rhythmicity of immune functions. In contrast, the present study suggests that pineal gland influence of immune function(s) is more specific and that pineal gland interaction with NK cells has a circadian component.

Correlations of serotonin and its metabolites in individual rat pineal glands over light: Dark cycles and after acute light exposure

Profiles of pineal indolealkylamines were estimated by high performance liquid chromatography and were correlated in individual glands of male rats sacrificed over several light:dark cycles and after acute exposure to light at night. A significant and positive correlation of 5HIAA vs 5HT in individual glands over both normal and experimental lighting conditions suggested that oxidative deamination is not a major factor in photic regulation of pineal 5HT levels and that the formation of 5HIAA is dependent on substrate availability. Regression analysis of other indole constituents revealed that there was a positive and significant correlation between 5HT vs N-acetylserotonin, but not between 5HT vs melatonin and N-acetylserotonin vs melatonin in individual glands during the dark phase of a light:dark cycle. We propose that this effect may be related to a pulsatile release of melatonin into the blood stream and is the result of sampling glands at different stages in the storage/release of melatonin.

The pineal of the troglophilic fish, Chologaster agassizi: an ultrastructural study.

The pineal of the facultative, cave-dwelling fish,Chologaster agassizi, was examined electron microscopically. Two cell types, photoreceptor and supportive cells, were identified in the pineal epithelium. The photoreceptor cells had well developed outer segments and contained Golgi bodies which were surrounded by both clear and dense-cored vesicles. Vesicle-crowned rods were frequently seen in various regions of the cell. The supportive cells also contained Golgi bodies from which both clear and dense-cored vesicles appeared to originate. In addition, these cells were characterized by peculiar arrangements of the smooth endoplasmic reticulum and the presence of pigment granules. Large quantities of glycogen were observed in both cell types. Small, unmyelinated nerve fibers were seen coursing throughout the pineal epithelium. Terminals filled with pleomorphic, clear vesicles and dense-cored vesicles were present in the vicinity of these nerve fibers. Similar vesicle-filled terminals were observed in close association with the supportive cells. The results of this study indicate that the pineal in this light-deprived species is a metabolically active organ capable of photoreception. Specializations of the organelles in the pineal cells were similar to those observed in other vertebrates living in environments of low light levels.