A retrograde double-labeling technique for light microscopy A combination of axonal transport of cholera toxin B-subunit and a gold-lectin conjugate

A light microscopical, non-fluorescent, retrograde double-labeling technique is described. Cholera toxin B-subunit (CTb) and a conjugate of wheatgerm agglutinin and bovine serum albumin coupled to 10 nm gold particles (gold-lectin) are both excellent retrograde tracers and, when visualized by means of immunohistochemistry and silver intensification, respectively, may be readily identified within the same cell. This light microscopical retrograde double-labeling technique is illustrated in rat with experiments designed to investigate the collat

Ursinus College Alumni Journal, November 1965

First words • From the President • Planning for the future at Ursinus College • Start of a four-year celebration • Guide to Centennial giving • Soviet portfolio: Rise and fall of the literary temperature; Evgeny Yevtushenko, Siberian pastorale; Memories of a Russian honeymoon • Founders Day is Ladies Day • Alumni come home to Ursinus • The many lives of Wismer Hall • Matching gifts lift industry & academia • Found: Preacher Mack • On Peace Corps duty in Thailand • The war trap • Lengthening list • Sporting scene • Campus clippings • Letters • Profiles: Four Ursinus men of distinction • Class notebook • Weddings • Births • In memoriam • A tribute to J. Allen Minnich • End quotes: The measles principle of educationhttps://digitalcommons.ursinus.edu/alumnijournal/1083/thumbnail.jp

Ins and Outs of Cerebellar Modules

The modular concept of cerebellar connections has been advocated in the lifetime work of Jan Voogd. In this concept, a cerebellar module is defined as the conglomerate of one or multiple and non-adjacent, parasagittally arranged zones of Purkinje cells, their specific projection to a well-defined region of the cerebellar nuclei, and the climbing fiber input to these zones by a well-defined region of the inferior olivary complex. The modular organization of these olivo-cortico-nuclear connections is further exemplified by matching reciprocal connections between inferior olive and cerebellar nuclei. Because the different regions of the cerebellar nuclei show highly specific output patterns, cerebellar modules have been suggested to constitute functional entities. This idea is strengthened by the observation that anatomically defined modules adhere to the distribution of chemical markers in the cerebellar cortex suggesting that modules not only differ in their input and output relations but also may differ in operational capabilities. Here, I will briefly review some recent data on the establishment of cerebellar modules in rats. Furthermore, some evidence will be shown suggesting that the other main afferent system (i.e., mossy fibers), at least to some extent, also adheres to the modular organization. Finally, using retrograde transneuronal tracing with rabies virus, some evidence will be provided that several cerebellar modules may be involved in the control of individual muscles

Cerebral activations related to ballistic, stepwise interrupted and gradually modulated movements in parkinson patients

Patients with Parkinson's disease (PD) experience impaired initiation and inhibition of movements such as difficulty to start/stop walking. At single-joint level this is accompanied by reduced inhibition of antagonist muscle activity. While normal basal ganglia (BG) contributions to motor control include selecting appropriate muscles by inhibiting others, it is unclear how PD-related changes in BG function cause impaired movement initiation and inhibition at single-joint level. To further elucidate these changes we studied 4 right-hand movement tasks with fMRI, by dissociating activations related to abrupt movement initiation, inhibition and gradual movement modulation. Initiation and inhibition were inferred from ballistic and stepwise interrupted movement, respectively, while smooth wrist circumduction enabled the assessment of gradually modulated movement. Task-related activations were compared between PD patients (N = 12) and healthy subjects (N = 18). In healthy subjects, movement initiation was characterized by antero-ventral striatum, substantia nigra (SN) and premotor activations while inhibition was dominated by subthalamic nucleus (STN) and pallidal activations, in line with the known role of these areas in simple movement. Gradual movement mainly involved antero-dorsal putamen and pallidum. Compared to healthy subjects, patients showed reduced striatal/SN and increased pallidal activation for initiation, whereas for inhibition STN activation was reduced and striatal-thalamo-cortical activation increased. For gradual movement patients showed reduced pallidal and increased thalamo-cortical activation. We conclude that PD-related changes during movement initiation fit the (rather static) model of alterations in direct and indirect BG pathways. Reduced STN activation and regional cortical increased activation in PD during inhibition and gradual movement modulation are better explained by a dynamic model that also takes into account enhanced responsiveness to external stimuli in this disease and the effects of hyper-fluctuating cortical inputs to the striatum and STN in particular

Behavioural Significance of Cerebellar Modules

A key organisational feature of the cerebellum is its division into a series of cerebellar modules. Each module is defined by its climbing input originating from a well-defined region of the inferior olive, which targets one or more longitudinal zones of Purkinje cells within the cerebellar cortex. In turn, Purkinje cells within each zone project to specific regions of the cerebellar and vestibular nuclei. While much is known about the neuronal wiring of individual cerebellar modules, their behavioural significance remains poorly understood. Here, we briefly review some recent data on the functional role of three different cerebellar modules: the vermal A module, the paravermal C2 module and the lateral D2 module. The available evidence suggests that these modules have some differences in function: the A module is concerned with balance and the postural base for voluntary movements, the C2 module is concerned more with limb control and the D2 module is involved in predicting target motion in visually guided movements. However, these are not likely to be the only functions of these modules and the A and C2 modules are also both concerned with eye and head movements, suggesting that individual cerebellar modules do not necessarily have distinct functions in motor control

Characterization of cytochrome P450 monooxygenase CYP154H1 from the thermophilic soil bacterium Thermobifida fusca

Cytochrome P450 monooxygenases are valuable biocatalysts due to their ability to hydroxylate unactivated carbon atoms using molecular oxygen. We have cloned the gene for a new cytochrome P450 monooxygenase, named CYP154H1, from the moderately thermophilic soil bacterium Thermobifida fusca. The enzyme was overexpressed in Escherichia coli at up to 14% of total soluble protein and purified to homogeneity in three steps. CYP154H1 activity was reconstituted using putidaredoxin reductase and putidaredoxin from Pseudomonas putida DSM 50198 as surrogate electron transfer partners. In biocatalytic reactions with different aliphatic and aromatic substrates of varying size, the enzyme converted small aromatic and arylaliphatic compounds like ethylbenzene, styrene, and indole. Furthermore, CYP154H1 also accepted different arylaliphatic sulfides as substrates chemoselectively forming the corresponding sulfoxides and sulfones. The enzyme is moderately thermostable with an apparent melting temperature of 67°C and exhibited still 90% of initial activity after incubation at 50°C

Pre-pregnancy body mass index and gestational weight gain and their effects on pregnancy and birth outcomes: a cohort study in West Sumatra, Indonesia

Background: Indonesia has a considerably high incidence of maternal and infant mortality. The country has however been experiencing a social and economic transition, influencing its general population demographics and nutritional status including the state of health and nutrition of pregnant women. This study aimed to explore body mass index (BMI) and gestational weight gain (GWG), and their relationship with pregnancy outcomes in a sample of Indonesian pregnant women. Methods: This observational cohort study included a total of 607 pregnant women who were recruited in 2010 from maternity clinics in Western Sumatra, Indonesia. Multiple logistic and regression analyses were undertaken to compare pregnancy and birth outcomes for different BMI and GWG, using normal weight women and women with a recommended weight gain as the referent groups. Results: The prevalence of underweight (BMI < 18.5 kg/m2) in pregnancy was high at 20.1%; while 21.7% of women were overweight (BMI: 23.0–27.4 kg/m2) and 5.3% obese (BMI ≥ 27.5 kg/m2) using the Asian BMI classifications. The incidence of overweight (BMI: 25.0–29.9 kg/m2) and obese (BMI ≥ 30.0 kg/m2) according to the international BMI classifications were 13.5% and 1.1% respectively. The majority of women gained inadequate weight in pregnancy compared to the Institute of Medicine (IOM)recommendations, especially those who had a normal BMI. Birthweight adjusted mean difference aMD (95% confidence interval) 205 (46,365) and the odds of macrosomia adjusted odds ratio aOR 13.46 (2.32–77.99) significantly increased in obese women compared to those with a normal BMI. Birthweight aMD -139 (−215, −64) significantly decreased in women with inadequate GWG compared to those with recommended GWG, while SGA aOR 5.44 (1.36, 21.77) and prematurity aOR 3.55 (1.23, 10.21) increased. Conclusions: Low nutritional status and inadequate GWG remain a cause for concern in these women. The higher odds of macrosomia with increasing maternal BMI and higher odds of prematurity and small for gestational age infants with inadequate weight gain also require attention. Research and practice recommendations: Urgent attention is required by researchers, policy makers and decision makers to facilitate development of culturally sensitive interventions to enhance nutritional status and health of mothers and babies, in an area known for its high incidence of maternal and neonatal mortality. Keywords: Maternal BMI, Gestational weight gain, Pregnancy outcomes, Birthweight, Indonesia, Cohort stud

Purkinje cell input to cerebellar nuclei in tottering: Ultrastructure and physiology

Homozygous tottering mice are spontaneous ataxic mutants, which carry a mutation in the gene encoding the ion pore of the P/Q-type voltage-gated calcium channels. P/Q-type calcium channels are prominently expressed in Purkinje cell terminals, but it is unknown to what extent these inhibitory terminals in tottering mice are affected at the morphological and electrophysiological level. Here, we investigated the distribution and ultrastructure of their Purkinje cell terminals in the cerebellar nuclei as well as the activities of their target neurons. The densities of Purkinje cell terminals and their synapses were not significantly affected in the mutants. However, the Purkinje cell terminals were enlarged and had an increased number of vacuoles, whorled bodies, and mitochondria. These differences started to occur between 3 and 5 weeks of age and persisted throughout adulthood. Stimulation of Purkinje cells in adult tottering mice resulted in inhibition at normal latencies, but the activities of their postsynaptic neurons in the cerebellar nuclei were abnormal in that the frequency and irregularity of their spiking patterns were enhanced. Thus, although the number of their terminals and their synaptic contacts appear quantitatively intact, Purkinje cells in tottering mice show several signs of axonal damage that may contribute to altered postsynaptic activities in the cerebellar nuclei

Local Narrative-Making on Refugees: How the Interaction Between Journalists and Policy Networks Shapes the Media Frames

AbstractCity narratives are crucial in shaping public attitudes and perceptions, and in defining the viable policy options and cities' responses to hot issues, such as migration and asylum. Nevertheless, the literature on relations between media and political actors is scarce and often unable to account for the complex local mechanisms leading to the production of media frames. This chapter investigates two urban crises: the rapid increase of transit refugees at the Central Station of Milan, and refugees' illegal occupation of four buildings in the ex-MOI area (former Olympic village) in Turin. Both events started in 2013 and have been studied from their beginnings up to mid-2016 by using qualitative techniques and media frame analysis. By matching the media and policy analysis, the authors show the central role that the local institutions can play in shaping media narratives on migrants and how the cohesion of the policy networks strengthens their ability to affect the local media frames. The dynamics of local journalism also matter: the presence of reporters with specific expertise, a commitment to migration and stable engagement in the issue, as well as collaboration within and between newsrooms, namely the existence of a sort of local media community, contribute to the development of consistent narratives over time and the prevalence of humanitarian rather than security frames