The Duration of Increased Grain Feeding Affects the Microbiota throughout the Digestive Tract of Yearling Holstein Steers.

Effects of the duration of moderate grain feeding on the taxonomic composition of gastrointestinal microbiota were determined in 15 Holstein yearling steers. Treatments included feeding a diet of 92% dry matter (DM) hay (D0), and feeding a 41.5% barley grain diet for 7 (D7) or 21 d (D21) before slaughter. At slaughter, digesta samples were collected from six regions, i.e., the rumen, jejunum, ileum, cecum, colon, and rectum. Extracted DNA from these samples was analyzed using MiSeq Illumina sequencing of the V4 region of the 16S rRNA gene. Three distinct PCoA clusters existed, i.e., the rumen, the jejunum/ileum, and the cecum/colon/rectum. Feeding the grain diet for 7 d reduced microbial diversity in all regions, except the ileum. Extending the duration of grain feeding from 7 to 21 d did not affect this diversity further. Across regions, treatment changed the relative abundances of 89 genera. Most of the changes between D0 and D7 and between D7 and D21 were opposite, demonstrating the resilience of gastrointestinal microbiota to a moderate increase in grain feeding. Results show that the duration of a moderate increase in grain feeding affects how gastrointestinal microbiota respond to this increase.

Effect of increasing the proportion of dietary concentrate on gastrointestinal tract measurements and brush border enzyme activity in Holstein steers.

The aim of this study was to determine the time course for adaptation of the reticulo-rumen, omasum, abomasum, and small intestine in response to an abrupt increase in the proportion of grain in the diet. Adaptive responses include tissue and digesta mass, small intestinal length, and brush border enzyme activity in the duodenum, proximal jejunum, and ileum. Twenty-five Holstein steers (213 ± 23 kg; 5 to 7 mo of age) were blocked by body weight, and within block were randomly assigned to 1 of 5 treatments: the control diet (CTRL; 92% chopped grass hay and 8% mineral and vitamin supplement on a dry matter basis) or a moderate grain diet (MGD; 50% chopped grass hay, 42% rolled barley grain, and 8% mineral and vitamin supplement) that was fed for 3 (MGD3), 7 (MGD7), 14 (MGD14), or 21 d (MGD21). Dry matter intake was limited to 2.25% of body weight to ensure that changes in dry matter intake did not confound the results. On the last day of the dietary exposure, steers were slaughtered 2 h after feeding. Reticulo-rumen tissue mass and ruminal epithelium mass in the ventral sac of the rumen were not affected by the MGD. Wet reticulo-ruminal digesta mass decreased from CTRL to MGD7 and then increased, but reticulo-ruminal digesta dry matter mass did not differ between treatments. Omasal mass, omasal tissue mass, and omasum digesta mass decreased linearly with the number of days fed MGD, but abomasal tissue mass tended to increase linearly. Duodenal tissue mass tended to increase linearly, and ileal length increased linearly with the number of days fed MGD. Lactase activity in the proximal jejunum increased linearly and maltase activity in duodenum tended to increase linearly with days fed MGD. Aminopeptidase N activity in the proximal jejunum increased cubically with days fed MGD, and dipeptidylpeptidase IV activity in ileum tended to decrease from CTRL to MGD14 and then tended to increase. Adaptation to a diet with a greater proportion of concentrate involves changes in the mass and length of regions of the gastrointestinal tract and brush border enzyme activity. These changes take place gradually over at least 3 wk.

Serosal-to-mucosal urea flux across the isolated ruminal epithelium is mediated via urea transporter-B and aquaporins when Holstein calves are abruptly changed to a moderately fermentable diet.

Urea transport (UT-B) proteins are known to facilitate urea movement across the ruminal epithelium; however, other mechanisms may be involved as well because inhibiting UT-B does not completely abolish urea transport. Of the aquaporins (AQP), which are a family of membrane-spanning proteins that are predominantly involved in the movement of water, AQP-3, AQP-7, and AQP-10 are also permeable to urea, but it is not clear if they contribute to urea transport across the ruminal epithelium. The objectives of this study were to determine (1) the functional roles of AQP and UT-B in the serosal-to-mucosal urea flux (Jsm-urea) across rumen epithelium; and (2) whether functional adaptation occurs in response to increased diet fermentability. Twenty-five Holstein steer calves (n=5) were assigned to a control diet (CON; 91.5% hay and 8.5% vitamin and mineral supplement) or a medium grain diet (MGD; 41.5% barley grain, 50% hay, and 8.5% vitamin and mineral) that was fed for 3, 7, 14, or 21 d. Calves were killed and ruminal epithelium was collected for mounting in Ussing chambers under short-circuit conditions and for analysis of mRNA abundance of UT-B and AQP-3, AQP-7, and AQP-10. To mimic physiologic conditions, the mucosal buffer (pH 6.2) contained no urea, whereas the serosal buffer (pH 7.4) contained 1 mM urea. The fluxes of (14)C-urea (Jsm-urea; 26 kBq/10 mL) and (3)H-mannitol (Jsm-mannitol; 37 kBq/10 mL) were measured, with Jsm-mannitol being used as an indicator of paracellular or hydrophilic movement. Serosal addition of phloretin (1 mM) was used to inhibit UT-B-mediated urea transport, whereas NiCl2 (1 mM) was used to inhibit AQP-mediated urea transport. Across treatments, the addition of phloretin or NiCl2 reduced the Jsm-urea from 116.5 to 54.0 and 89.5 nmol/(cm(2) × h), respectively. When both inhibitors were added simultaneously, Jsm-urea was further reduced to 36.8 nmol/(cm(2) × h). Phloretin-sensitive and NiCl2-sensitive Jsm-urea were not affected by diet. The Jsm-urea tended to increase linearly as the duration of adaptation to MGD increased, with the lowest Jsm-urea being observed in animals fed CON [107.7 nmol/(cm(2) × h)] and the highest for those fed the MGD for 21 d [144.2 nmol/(cm(2) × h)]. Phloretin-insensitive Jsm-urea tended to increase linearly as the duration of adaptation to MGD increased, whereas NiCl2-insensitive Jsm-urea tended to be affected by diet. Gene transcript abundance for AQP-3 and UT-B in ruminal epithelium increased linearly as the duration of MGD adaptation increased. For AQP-7 and AQP-10, gene transcript abundance in animals that were fed the MGD was greater compared with that of CON animals. These results demonstrate that both AQP and UT-B play significant functional roles in urea transport, and they may play a role in urea transport during dietary adaptation to fermentable carbohydrates.

Effects of duration of moderate increases in grain feeding on endotoxins in the digestive tract and acute phase proteins in peripheral blood of yearling calves.

Effects of duration of grain feeding on the concentration of endotoxic lipopolysaccharide (LPS) in digesta throughout the digestive tract and on acute phase proteins and LPS in peripheral blood were determined in Holstein yearling calves. Twenty-five Holstein yearling steer calves received either a forage-based diet containing 92% hay and 8% of a mineral and vitamin pellet on a dry matter basis (CON) or a moderate-grain diet, obtained by replacing 41.5% of the hay in the forage-based diet with barley grain, for 3 (MG3), 7 (MG7), 14 (MG14), or 21 d (MG21) before slaughter. Immediately before slaughter, blood samples were collected from the jugular vein. Immediately after slaughter, digesta samples were collected from the rumen, jejunum, ileum, cecum, colon, and rectum. Rumen liquid digesta, digesta from the intestines, and peripheral blood plasma were analyzed for LPS. Peripheral blood plasma and serum were analyzed for the acute phase proteins amyloid A, haptoglobin, and LPS-binding protein. Feeding the grain diet increased the LPS concentration in rumen fluid linearly from 15,488 endotoxin units (EU)/mL for CON to 70,146 EU/mL for MG7. Concentrations of LPS in rumen fluid in MG14 and MG21 were 61,944 and 56,234 EU/mL, respectively, and did not differ. The LPS concentrations in jejunal digesta were much lower than that in digesta elsewhere in the digestive tract, which suggests that ruminal LPS is broken down in the abomasum or proximal jejunum. The concentration of digesta LPS in the ileum was higher than that of digesta elsewhere in the intestines and similar to that in rumen fluid. The duration of grain feeding increased the LPS concentration in digesta in the ileum and cecum and tended to increase that in the colon cubically. Concentrations of LPS in this part of the digestive tract were highest in the MG3 and MG21 groups. The highest concentrations of LPS in digesta in the cecum, colon, and rectum were 3.7, 3.8, and 5.6 times higher than that in CON, respectively. Grain feeding and the increase in LPS in digesta were not accompanied by an acute phase response or a detectable concentration of LPS in peripheral blood. The absence of LPS in peripheral blood and the lack of increase in acute phase proteins indicated that the grain feeding protocol used in the current study and the accompanying changes in LPS concentrations of the digesta did not result in systemic inflammation.

Fate of pharmaceuticals and bacteria in stored urine during precipitation and drying of struvite.

Experiments were conducted to measure the behaviour of eight pharmaceuticals during urine treatment as part of the project 'SANIRESCH - Sustainable sanitary recycling Eschborn'. Urine was collected from 200 people in a public building via waterless urinals and NoMix toilets. It was then stored at room temperature at different pH values to analyse the extent to which bacteria and pharmaceuticals are eliminated over time. Although a partial elimination of pharmaceuticals could be detected, the storage at defined pH values cannot be advised. As the persons tested used pharmaceuticals with different structures, in different amounts and at varying intervals, this method of treatment is insufficient for removing them from urine. Precipitating the urine with MgO, washing it with saturated struvite solution and drying it at 30 °C will result in a free-flowing granular powder of struvite (NH(4)MgPO(4)·6H(2)O) that is free of pharmaceuticals and pathogens and can be used as fertiliser and a source of nitrogen, magnesium and phosphorus.

A hierarchical neural system with attentional top-down enhancement of the spatial resolution for object recognition.

We present a hierarchical neurodynamical system for object recognition based on attentional control of the spatial resolution with which an object is analyzed during an iterative hypothesis testing cycle. Psychophysical evidence strongly suggests that attentional processing results in the enhancement of the spatial resolution in the input region corresponding to the focus of attention. We adopt a computational neuroscience approach in order to analyze this attentional enhancement of the spatial resolution for object recognition. The system consists of a where- and a what-module which include networks with feedforward and feedback interconnections describing the mutual links between different areas of the visual cortex.

A neuro-cognitive visual system for object recognition based on testing of interactive attentional top-down hypotheses.

We propose an extension of a systemic model for object recognition formulated by Rybak et al (1998 Vision Research 38 2387-2400) which is based on the functional organisation of the visual systems in primate brains. In contrast to the learning and recognition scheme of Rybak et al we do not assume a behavioural paradigm, i.e. a visuomotor programmed scanpath that determines the sequence of foveation on the different parts of the object. As in the basis architecture of Rybak et al, the system modules are separated into 'what'-like subsystems corresponding to the ventral occipito-inferotemporal visual path and 'where'-like complexes analogous to the dorsal occipito-parietal visual path. The 'what' system analyses local features in the actual foveation as in Rybak et al. But, in our case, the 'where' memory, instead of programming a behavioural scanpath, scores the spatial relationship between successive fixation and the spatial relationship between the associated main edges. The recognition is based on the identification of parts and their spatial relationship. This gives the learning and recognition mechanisms more flexibility in the sense that, for recognising an object, several different fixation sequences may be accepted.

Spatiotemporal coding in the cortex: information flow-based learning in spiking neural networks.

We introduce a learning paradigm for networks of integrate-and-fire spiking neurons that is based on an information-theoretic criterion. This criterion can be viewed as a first principle that demonstrates the experimentally observed fact that cortical neurons display synchronous firing for some stimuli and not for others. The principle can be regarded as the postulation of a nonparametric reconstruction method as optimization criteria for learning the required functional connectivity that justifies and explains synchronous firing for binding of features as a mechanism for spatiotemporal coding. This can be expressed in an information-theoretic way by maximizing the discrimination ability between different sensory inputs in minimal time.

The coding of information by spiking neurons: an analytical study.

We analyse analytically the coding of information by a spiking neuron. The emphasis is on the question of how many spikes are necessary for the reliable discrimination of two different input signals. The discrimination ability is measured by the second-order Rényi mutual information between the random variable describing the name of the signal and a sequence of n output spikes. Analysing this measure as a function of n, we study the coding strategy of a single spiking neuron, with the following main results. A small number of output spikes is required for efficient discrimination of input signals, i.e. for encoding them, if the separation is easy; a large number of output spikes is required in the difficult case of separation of very similar input signals. Three different versions of the spike response model of a single neuron are studied. The approach presented can be regarded as a non-parametric version of the reconstruction method of Bialek.

Differential modulation of AMPA receptor mediated currents by evans blue in postnatal rat hippocampal neurones.

1. The modulation of non-N-methyl-D-aspartate (NMDA) receptor-mediated whole cell currents and of glutamatergic synaptic transmission by purified Evans Blue (EB) was investigated in rat cultured postnatal hippocampal neurones by use of patch clamp recordings and a fast drug application system. 2. Three different groups of neurones could be distinguished with respect to the type of modulation obtained with 10 microM EB: EB was either a predominant inhibitor of desensitization (13% of the neurones), a predominant inhibitor of current amplitudes (42%) or a mixed inhibitor of both properties (45%). Both effects were not use-dependent and reached maximal levels after 30 s of pre-equilibration with the diazo dye. 3. Dose-response curves obtained from glutamate activated whole cell currents yielded an IC50 value for EB of 13.3 microM (Hill coefficient: 1.3) for the inhibition of desensitization, and an IC50 value of 10.7 microM (Hill coefficient: 1.2) for the inhibition of current amplitudes. 4. Chicago acid SS (100 microM) which is one of the synthesis precursors of EB had no effect on current amplitudes of glutamate activated whole cell currents but was a weak inhibitor of desensitization in all hippocampal neurones investigated, irrespective of the type of modulation obtained with EB in the same neurone. 5. Oxidatively modified EB (the so-called VIMP (10 microM)) had no effect on the kinetics but was a partial inhibitor of glutamate-activated whole cell currents in all hippocampal neurones investigated. 6. EB (10 microM) inhibited the amplitudes of non-NMDA receptor mediated autaptic currents to the same extent (to 39 +/- 19% of control) as observed for glutamate activated whole cell currents (to 41 +/- 17% and 56 +/- 20%). However, the decay of the autaptic responses remained uninfluenced upon EB application, indicating that either receptor desensitization does not dominate the time course of the synaptic response or that the non-NMDA receptors sensitive to modulation of desensitization by EB are not present in the postsynaptic membrane. 7. In conclusion, EB differentially modulates alpha-amino-3-hydroxy-5-methyl -4-isoxazole propionic acid (AMPA) receptor gating in different subsets of neurones. Upon identification of the cellular determinants for the differential modulation (e.g. AMPA receptor subunit composition) EB could become a useful tool to investigate receptor subtypes during electrophysiological recordings.

[Immunohistochemical demonstration of extracerebral toxoplasmosis in AIDS]. / Immunhistologischer Nachweis der extrazerebralen Toxoplasmose bei AIDS.

Tissue slides obtained at autopsy from 80 cases with AIDS were studied immunhistochemically for infection with Toxoplasma gondii. In 35 cases (43.75%) toxoplasmosis could be found: in 22 cases (27.5%) only cerebral, in 9 cases (11.25%) cerebral and extracerebral and in 4 cases (5%) only extracerebral. Necrotizing lesions, due to the parasite could be seen in brain, heart, lungs, pancreas, adrenal glands and testis, only intracellular trophozoites without tissue damage in GIT, liver, lymphnodes, spleen, prostate, kidney and gl. parotis. The trophozoites and pseudocysts could be clearly demonstrated by immunohistochemistry.

[Clinical significance of the short-term incubation test for the therapy of metastatic breast cancer]. / Klinische Bedeutung des Kurzzeitinkubationstests für die Therapie des metastasierten Mammakarzinoms.

Between January 1978 and October 1980 97 tissue samples of histologically verified carcinoma of the breast were received for performance of the short-term incubation test. 25 tumour samples could not be prepared as cell suspension sufficient for testing. Among the 72 performed tests there were 9 with stimulation of 3H-uridine uptake by doxorubicin. Thus only 63 tests could be evaluated. Results were correlated with clinical data of the patients. No significant correlations could be established between tumour stage at time of diagnosis, age, menopausal state, receptor state, and the test result. There was also no differentiation between favourable and unfavourable prognoses as regards free interval and rate of survival. A correlation between results of tests and success or failure of cytostatic treatment could not be ascertained.