High-yield and high-purity amide bond formation using DMTMM PF6 for DNA-encoded libraries.

In this study, we report on the ability of DMTMM PF6 to improve the amidation reaction. The on-DNA amidation reaction using DMTMM PF6 demonstrates higher conversion rates than those using HATU or DMTMM Cl, particularly with challenging sterically hindered amines and carboxylic acids. The developed method enables the expansion of available building blocks and the efficient synthesis of high-purity DNA-encoded libraries.

Motor skills training-induced activation of descending pathways mediating cortical command to hindlimb motoneurons in experimental diabetic rats.

Diabetes disrupts the corticospinal tract (CST) system components that control hindlimb and trunk movement, resulting in weakness of the lower extremities. However, there is no information about a method to improve these disorders. This study aimed to investigate the rehabilitative effects of 2 weeks of aerobic training (AT) and complex motor skills training (ST) on motor disorders in streptozotocin-induced type 1 diabetic rats. In this study, electrophysiological mapping of the motor cortex showed that the diabetes mellitus (DM)-ST group had a larger motor cortical area compared to the DM-AT group and sedentary diabetic animals. Moreover, hand grip strength and rotarod latency increased in the DM-ST group; however, these two parameters did not change in the DM-AT group, as well as in control and sedentary diabetic rats. Furthermore, in the DM-ST group, cortical stimulation-induced and motor-evoked potentials were preserved after the interception of the CST; however, this potential disappeared after additional lesions were made on lateral funiculus, suggesting that their function extends to activating motor descending pathways other than the CST locating lateral funiculus. According to immunohistochemical analysis, the larger fibers present on the dorsal part of the lateral funiculus, which corresponds to the rubrospinal tract of the DM-ST group, expressed the phosphorylated growth-associated protein, 43 kD, which is a specific marker of axons with plastic changes. Additionally, electrical stimulation of the red nucleus revealed expansion of the hindlimb-responsible area and increased motor-evoked potentials of the hindlimb in the DM-ST group, suggesting a strengthening of synaptic connections between the red nucleus and spinal interneurons driving motoneurons. These results reveal that ST induces plastic changes in the rubrospinal tract in a diabetic model, which can compensate for diabetes by disrupting the CST system components that control the hindlimb. This finding suggests that ST can be a novel rehabilitation strategy to improve motor dysfunctions in diabetic patients.

Functional and Structural Changes in the Corticospinal Tract of Streptozotocin-Induced Diabetic Rats.

This study aimed to reveal functional and morphological changes in the corticospinal tract, a pathway shown to be susceptible to diabetes. Type 1 diabetes was induced in 13-week-old male Wistar rats administered streptozotocin. Twenty-three weeks after streptozotocin injection, diabetic animals and age-matched control animals were used to demonstrate the conduction velocity of the corticospinal tract. Other animals were used for morphometric analyses of the base of the dorsal funiculus of the corticospinal tract in the spinal cord using both optical and electron microscopy. The conduction velocity of the corticospinal tract decreased in the lumbar spinal cord in the diabetic animal, although it did not decrease in the cervical spinal cord. Furthermore, atrophy of the fibers of the base of the dorsal funiculus was observed along their entire length, with an increase in the g-ratio in the lumbar spinal cord in the diabetic animal. This study indicates that the corticospinal tract fibers projecting to the lumbar spinal cord experience a decrease in conduction velocity at the lumbar spinal cord of these axons in diabetic animals, likely caused by a combination of axonal atrophy and an increased g-ratio due to thinning of the myelin sheath.

High-Fat Diet and Age-Dependent Effects of IgA-Bearing Cell Populations in the Small Intestinal Lamina Propria in Mice.

Several studies highlighted that obesity and diabetes reduce immune function. However, changes in the distribution of immunoglobins (Igs), including immunoglobulin-A (IgA), that have an important function in mucosal immunity in the intestinal tract, are unclear. This study aimed to investigate the impaired immune functions in the context of a diet-induced obese murine model via the assessment of the Igs in the intestinal villi. We used mice fed a high-fat diet (HFD) from four to 12 or 20 weeks of age. The distributions of IgA, IgM, and IgG1 were observed by immunohistochemistry. Interestingly, we observed that IgA was immunolocalized in many cells of the lamina propria and that immunopositive cells increased in mice aged 12 to 20 weeks. Notably, mice fed HFD showed a reduced number of IgA-immunopositive cells in the intestinal villi compared to those fed standard chow. Of note, the levels of IgM and IgG1 were also reduced in HFD fed mice. These results provide insights into the impaired mucosal immune function arising from diet-induced obesity and type 2 diabetes.

Properties of Renshaw-like cells excited by recurrent collaterals of pudendal motoneurons in the cat.

Although anatomical studies have indicated pudendal motoneurons to give off recurrent collaterals, they are not considered to make synapses onto interneurons, such as Renshaw cells, and rather terminate their own signals. No study till date has examined interneurons being driven by recurrent collaterals of pudendal motoneurons. Here, we aimed to investigate the existence of Renshaw cells driven by pudendal motoneurons along with the recurrent inhibition of the latter. Extracellular recordings were obtained from the ventral horn of the sacral spinal cord of anesthetized cats. Dorsal roots were sectioned, and motor axons were electrically stimulated. Renshaw-like cells driven by recurrent collaterals, with high-frequency firings at short latency discharge, were observed around Onuf's nucleus. However, the recurrent inhibitory post-synaptic potentials were not recorded by intracellular recordings from the pudendal motoneurons. In summary, we found Renshaw-like cells driven by pudendal motoneurons, but we could not identify the synaptic connection of these neurons.

Firing properties of medullary expiratory neurons during fictive straining in cats.

Expiratory (E) neurons in the caudal nucleus retroambigualis extend descending spinal axons to the lumbar and sacral spinal cord. Discharge rates of single E neurons were recorded to examine differences in activity of E neurons projecting to the lumbar or sacral spinal cord during fictive straining induced by distention of the colon with a balloon. Firing frequencies of E neurons with descending axons in the thoracic and lumbar spinal cord increased during the repetitive rise of rectum pressure, whereas those of E neurons with descending axons in the sacral spinal cord decreased. E neurons with descending axons in the thoracic/lumbar and sacral spinal cord exhibit different firing characteristics during the repetitive rise of rectum pressure when straining during defecation. The activity of abdominal nerves during fictive straining is in phase with changes in rectum pressure, but out of phase with the activity of E neurons.

Electrophysiological properties of Ia excitation and recurrent inhibition in cat abdominal motoneurons.

Ia excitation and recurrent inhibition are basic neuronal circuits in motor control in hind limb. Renshaw cells receive synaptic inputs from axon collaterals of motoneurons and inhibit motoneurons and Ia inhibitory interneurons. It is important to know properties of Ia excitation and recurrent inhibition of trunk muscle such as abdominal muscles. The abdominal muscles have many roles and change those roles for different kind of functions. Intracellular recordings were obtained from the abdominal motoneurons of the upper lumbar segments in cats anesthetized. First, dorsal roots were left intact, and sensory and motor axons were electrically stimulated. Ia excitatory post-synaptic potentials were elicited in five of eight motoneurons at same segment stimulated. Second, dorsal roots were sectioned, and motor axons were electrically stimulated. Recurrent inhibitory post-synaptic potentials were elicited in one of 11 abdominal motoneurons. Renshaw cells extracellularly fired high-frequency bursts at short latency and at same segment stimulated.

Effects of streptozotocin-induced diabetes on leg muscle contractile properties and motor neuron morphology in rats.

Skeletal muscle fiber subtypes are differentially sensitive to diabetes-related pathology; For example, fast-twitch muscles exhibit severe decreases in contraction force while slow-twitch muscles demonstrate prolonged half-relaxation time. However, such alterations have only been examined after a relatively short period following diabetes onset, with no information available regarding muscle damage caused by longer disease periods (>20 weeks). This study examined alterations in the contractile properties of the medial gastrocnemius (fast-twitch) and soleus (slow-twitch) muscles, as well as morphological changes in their motor neurons 12 and 22 weeks after diabetes onset. Adult male Wistar rats were divided into diabetic (12- or 22-week post-streptozotocin injection) and age-matched control groups. Electrically evoked maximum twitch and tetanic tension were recorded from leg muscles. Additionally, motor neuron number and cell body size were examined. At 12 weeks after diabetes onset, decreases in twitch force were observed predominantly in medial gastrocnemius muscles, while soleus muscles exhibited prolonged half-relaxation time. However, these differences became ambiguous at 22 weeks, with decreased twitch force and prolonged half-relaxation time observed in both muscles. On the other hand, reduction in soleus motor neurons was observed 12 weeks after diabetes onset, while medial gastrocnemius motor neurons were diminished at 22 weeks. These data indicate that experimental diabetes induces differential damage to medial gastrocnemius and soleus muscles as well as motor neurons. These diabetes-induced differences may partly underlie the differential deficits observed in gastrocnemius and soleus.

Effect of streptozotocin-induced diabetes on motor representations in the motor cortex and corticospinal tract in rats.

Motor disorders in patients with diabetes are associated with diabetic peripheral neuropathy, which can lead to symptoms such as lower extremity weakness. However, it is unclear whether central motor system disorders can disrupt motor function in patients with diabetes. In a streptozotocin-induced rat model of type 1 diabetes, we used intracortical microstimulation to evaluate motor representations in the motor cortex, recorded antidromic motor cortex responses to spinal cord stimulation to evaluate the function of corticospinal tract (CST) axons, and used retrograde labeling to evaluate morphological alterations of CST neurons. The diabetic rats exhibited size reductions in the hindlimb area at 4 weeks and in trunk and forelimb areas after 13 weeks, with the hindlimb and trunk area reductions being the most severe. Other areas were unaffected. Additionally, we observed reduced antidromic responses in CST neurons with axons projecting to lumbar spinal segments (CST-L) but not in those with axons projecting to cervical segments (CST-C). This was consistent with the observation that retrograde-labeled CST-L neurons were decreased in number following tracer injection into the spinal cord in diabetic animals but that CST-C neurons were preserved. These results show that diabetes disrupts the CST system components controlling hindlimb and trunk movement. This disruption may contribute to lower extremity weakness in patients.

Effect of streptozotocin-induced diabetes on motoneurons and muscle spindles in rats.

This study examined the alterations in the number and size of motoneurons innervating the medial gastrocnemius (MG) and biceps femoris (BF) motor nuclei in diabetic rats (12 or 22 weeks after injection of streptozotocin) and age-matched controls using retrograde labeling technique. Additionally, morphological alterations of muscle spindles in BF and MG muscles were tested. Significantly fewer labeled MG motoneurons were found in 12- and 22-week diabetic rats as compared with age-matched control animals. In contrast, the number of BF motoneurons was preserved in each group. Compared to control animals, the ratio of larger motoneurons of MG and BF muscle were decreased at 12 weeks, and smaller MG motoneurons were drastically decreased at 22 weeks. Moreover, MG muscle spindle showed reduction of its number and increase of intrafusal muscle fibers; however, BF muscle spindles showed little or no difference from control animals. We conclude that there is an early loss of alpha motoneurons for both MG and BF muscles followed by a later loss of gamma motoneurons in MG muscle in diabetic animals. Moreover, loss of gamma motoneuron might induce atrophy of MG muscle spindles.

Discharge patterns of abdominal and pudendal nerves during induced defecation in anesthetized cats.

Defecation is thought to be achieved not only by contraction of the colon, but also by a rise in intra-abdominal pressure. In this study we recorded the discharges of nerves innervating the abdominal (Abd) muscles, diaphragm, external anal sphincter (EAS) muscle and pelvic floor (PF) muscles during induced defecation evoked by distention of an expellable balloon to reveal defecation-related muscle activities. The discharges of the Abd muscle and phrenic (Phr) nerves increased when rectal pressure increased. The discharges of the EAS and PF nerves usually increased in proportion to the pressure in the rectum and maintained a constant activity level, although some trials showed inhibition. The results suggest that activities of these muscles increase the intra-abdominal pressure.

The size of motoneurons of the gastrocnemius muscle in rats with diabetes.

Alterations in the number and size of motoneurons were studied in the medial gastrocnemius (MG) motor nucleus of diabetic rats (12 or 22 weeks after injection of storeptozotocin) and age-matched controls. Each group contained 6 animals. MG motoneurons were retrogradely labeled by dextran-fluorescein and the number and size of cell bodies were examined. Significantly fewer labeled MG motoneurons were found in the 22-week diabetic rats as compared with age-matched control animals. The mean soma diameter of MG motoneurons was significantly smaller in the 12- and 22-week diabetic animals. Furthermore the soma size for 22-week diabetic animals was smaller than for 12-week diabetic animals. The distribution of average soma diameters in the MG nucleus of control animals was bimodal; cells with larger average diameter were presumed to be alpha-motoneurons and those with smaller diameters were presumed to be gamma. Compared to control animals, the number of smaller MG motoneurons was reduced in 12 week diabetic animals. By 22 weeks, diabetic animals had no small MG motoneurons and the size distribution became unimodal. We conclude that there is a significant decrease in the absolute number and size of MG motoneurons in diabetic rats, with the possibility that the decrease occurred predominantly among the smaller gamma-motoneurons.

Synthesis of a stable 1,2-bis(ferrocenyl)diphosphene.

The synthesis and characterization of a stable 1,2-bis(ferrocenyl)diphosphene, wherein a P=P π-bond connects two ferrocenyl units will be reported. This represents an unprecedented example for a d-π electron system containing a heavier pnictogen π-spacer group. Stabilization of the highly reactive P=P π-bond was achieved by steric protection using two bulky ferrocenyl moieties.

The diaphragmatic activities during trunk movements.

In order to investigate how the diaphragm and trunk muscles are recruited during various voluntary movements, we recorded EMG signals and video images simultaneously and analyzed EMGs of the diaphragm and trunk muscles during the voluntary movements that required trunk muscles. During trunk movements, the duration of the diaphragmatic activity became irregular and the intensity of the activity increased. Further analysis revealed that the diaphragmatic activities were consisted of two components, such as respiratory and non-respiratory activities during voluntary movements. Our results led to the idea that the diaphragmatic activities may be controlled from different control mechanisms of central nervous system.

Activity patterns of the diaphragm during voluntary movements in awake cats.

The diaphragm is an important inspiratory muscle, and is also known to participate in the postural function. However, the activity of the diaphragm during voluntary movements has not been fully investigated in awake animals. In order to investigate the diaphragmatic activity during voluntary movements such as extending or rotating their body, we analyzed the electromyogram (EMG) of the diaphragm and trunk muscles in the cat using a technique for simultaneous recordings of EMG signals and video images. Periodic respiratory discharges occurred in the left and right costal diaphragm when the cat kept still. However, once the cat moved, their periodicity and/or synchrony were sometimes buried by non-respiratory activity. Such non-periodic diaphragmatic activities during voluntary movements are considered as the combination of respiratory activity and non-respiratory activity. Most of the diaphragmatic activities started shortly after the initiation of standing-up movements and occurred after the onset of trunk muscle activities. Those activities were more active compared to the normal respiratory activity. During rotation movements, left and right diaphragmatic activities showed asymmetrical discharge patterns and higher discharges than those during the resting situation. This asymmetrical activity may be caused by taking different lengths of each side of the diaphragm and trunk muscles. During reaching movements, the diaphragmatic activity occurred prior to or with the onset of trunk muscle activities. It is likely that diaphragmatic activities during reaching movements and standing-up movements may have been controlled by some different control mechanisms of the central nervous system. This study will suggest that the diaphragmatic activity is regulated not only by the respiratory center but also by inputs from the center for voluntary movements and/or sensory reflex pathways under the awake condition.

Morphological study of external oblique motor nerves and nuclei in cats.

In order to clarify the morphological features of peripheral motor nerves and motoneurons that innervate trunk muscles, the size distribution of external oblique (EO) peripheral motor fibers and motoneurons of the thoracic and the lumbar segments were examined. Histograms of the size distribution of EO motor fibers in peripheral nerves after ganglionectomy clearly had a bimodal distribution of small fiber groups and large fiber groups. It is very likely that small fiber groups correspond to gamma motor fibers and large fiber groups to alpha motor fibers. Gamma and alpha motor fiber groups were separated at 8-14 microm. The average diameter of the gamma and alpha motor fibers were different in each segment. The ratio of gamma and alpha motor fibers was approximately 1:2.0 in the thoracic segments and from 1:1.8 to 1:0.9 in the lumbar segments. Horseradish peroxidase was applied to the central stump of EO nerves, and the size distribution of EO motoneuron cell bodies in the thoracic and the lumbar spinal cords was examined. The size distribution of motoneuron cell bodies was bimodal in one cat (small and large motoneurons) and unimodal in three cats. When the ratio of small motor fibers to large motor fibers in peripheral nerves was applied to that of small motoneurons to large motoneurons, the separation of small and large motoneurons was approximately 40 microm. These results suggest that the morphological characteristics in peripheral nerves of trunk muscles are not reflected in motoneurons.

Zwitterionic salts as mild organocatalysts for transesterification.

The exothermic reaction of 3,5-bis(trifluoromethyl)phenyl or 4-nitrophenyl isothiocyanate with 4-pyrrolidinopyridine (PPY) gave the corresponding arylaminothiocarbonylpyridinium salts in quantitative yields. These novel zwitterionic salts were effective as organocatalysts for the transesterification reaction of an equimolar mixture of methyl carboxylates and alcohols in hydrocarbons such as heptane and octane under azeotropic reflux conditions with the removal of methanol. In sharp contrast, PPY was inert as a catalyst under the same reaction conditions.

Vertical eye movement-related type II neurons with downward on-directions in the vestibular nucleus in alert cats.

The vestibular nuclei and the interstitial nucleus of Cajal (INC) have been regarded as key elements of the velocity-to-position integrator for vertical eye movements. This paper reports a class of type II vestibular neurons that receives input from the INC and carries vertical eye movement signals that appear to represent an intermediate stage of the integration process. Extracellular recordings were made from neurons in and near the vestibular nuclei in alert cats. We encountered 39 neurons that exhibited an intense burst of spikes for downward saccades and a position-related tonic activity during intersaccadic intervals (d-type II neurons). They had a very high saccadic sensitivity (4.3+/-2.7 spikes/deg, mean +/- SD) as well as a high position sensitivity (3.2+/-1.6 (spikes/sec)/deg). Unlike the bursts of motoneurons, the bursts of these neurons declined gradually with an exponential-like time course and lasted well beyond the end of saccades. The mean time constant of the burst decay was 139+/-43 ms. The d-type II neurons were excited with disynaptic or trisynaptic latencies following stimulation of the contralateral vestibular nerve. The responses to vertical head rotations suggested inputs from the contralateral posterior canal. The d-type II neurons were excited with short latencies following stimulation of the ipsilateral INC, suggesting that they receive a direct excitatory input from vertical eye movement-related INC neurons with downward on-directions. The d-type II neurons were located in the rostral portion of the vestibular nuclei and the underlying reticular formation. These results suggest that d-type II neurons may be interposed between the burst-tonic neurons in the INC and pure tonic neurons in the vestibular nuclei and contribute to the oculomotor velocity-to-position integration.