Identification of candidate genes associated with resistance against race 0 of Colletotrichum lentis in Lens ervoides.

Resistance to anthracnose caused by the fungal pathogen Colletotrichum lentis was explored through transcriptome sequencing over a period of 24 to 96 h post-inoculation (hpi) of the partially resistant recombinant inbred lines (RIL) LR-66-528 and susceptible LR-66-524 of the crop wild relative Lens ervoides population LR-66. The development of infection vesicles and primary hyphae by C. lentis were significantly higher on susceptible RIL LR-66-524 compared to partially resistant LR-66-528 at 24 and 48 hpi, but exponential trends in fungal growth were observed between 24 to 96 hpi in both RILs. Comparison of inoculated with mock-inoculated samples revealed 3091 disease responsive genes, among which 477 were differentially expressed between the two RILs. These were clustered into six expression clusters with genes that had either high or low expression in one of the RILs. Differentially expressed genes (DEGs) were functionally annotated and included genes coding LRR and NB-ARC domain disease resistance proteins, protein detoxification, LRR receptor-like kinase family proteins, and wall-associated Ser/Thr Kinases. DEGs were compared to genes in previously published anthracnose resistance QTLs mapped in LR-66 and revealed 22 DEGs located in 3 QTLs. Expression of 21 DEGs was validated using RT-qPCR confirming expression trends in RNA-seq.

Persistent Organic Pollutants Residues in Human Breast Milk from Bathinda and Ludhiana Districts of Punjab, India.

In the present study, persistent organic pollutants (POPs), including six congeners of polychlorinated biphenyls (PCBs) and organochlorine pesticides, were estimated in human breast milk samples collected from two districts of Punjab (India). The mean concentrations of POP residues were recorded higher in Bathinda district (PCBs: 33.7; DDTs: 519.2; HCHs: 46.6 ng g-1 lipid wt.) than Ludhiana (PCBs: 24.2; DDTs; 415.3; HCHs; 35.5 ng g-1 lipid wt.). Levels of PCBs and DDTs were observed higher in primiparas, whereas HCHs residues were seen more in multiparas. Risk analysis to POPs exposure through breast milk reflected that the daily intake for some infants was close to or above the tolerable daily intake limit for detrimental effects, which may raise a health concern. Comparative evaluation of present data indicated that DDT and HCHs residue levels in human breast milk from Punjab, India were among the lowest values reported for developing countries. The first-order kinetic reaction at a steady-state condition used to estimate the half-life of DDT and HCH suggested that DDT levels have declined from 18,211 to 490 ng g-1 lipid wt. with a half-life (Tdec1/2) of 3.25 years over a span of 15 years. Similarly, HCH levels have decreased from 8609 to 46.6 ng g-1 in this duration with Tdec1/2 of 2.25 years. Because some infants are still at risk, continuous monitoring of POPs in human milk is needed for surveillance and interpretation of time trends and for linkage to strict enforcement of agricultural regulations.

Reciprocal inhibition versus unloading response during stretch reflex in humans.

Rotation of an upper limb joint produces excitatory stretch reflex peaks M1 and M2 in the stretched muscles and simultaneous decrease in electromyographic (EMG) activity in the shortened muscles. The objective of this study was to examine whether the decreased activity in the antagonists (rINHIB) is purely from unloading of the spindles or receives active inhibition involving inhibitory interneurons. If rINHIB is due only to unloading, then the termination of rINHIB should vary with the duration of perturbation used to elicit stretch reflex, namely shorter stretches should result in shorter values of decreased periods of EMG. To examine this question, rectangular pulses, ranging in duration from 25 to 150 ms, were used to stretch wrist flexors or extensors with a torque motor. These rectangular pulses resulted in joint rotations which peaked at times (T(peak)) ranging from approximately 75-160 ms. As shown by previous authors, when the duration of rotation was shortened, the magnitude of M1 did not change, while the magnitude of M2 decreased. However, termination time of rINHIB in the shortened muscles did not change with change in T(peak), implying thereby that unloading of spindles of the antagonist muscles is not the only mechanism for the reduction in activity and that inhibitory reflex pathways most likely contribute. Possible sources of inhibition are discussed for the short- and long-latency inhibition.

Proprioceptive reaction times and long-latency reflexes in humans.

The stretch of upper limb muscles results in two electromyographic (EMG) peaks, M1 and M2. The amplitude of M2 peak can generally be modified by giving prior instruction to the subject on how to react to the applied perturbation. The unresolved question is whether the amplitude modulation results from change in the gain of the reflex pathway contributing to M2, or by superposition of reaction time (RT) activity. The following study attempted to resolve this question by examining the overlap between proprioceptive RT and M2 activities. Subject's right wrist flexors were stretched, and he/she was instructed either (1) not to intervene (passive task) or (2) to react as fast as possible by simultaneously flexing both wrists (active or compensate task). Under passive and active conditions, M1 and M2 were observed from EMG of right wrist flexors, and during the active condition, RT activities were additionally observed from both sides. The onset and offset of M2 (M1(onset), M2(offset)) were measured from the passive averages, while the RT was measured from the averaged EMG response of the left wrist flexors. For between-subject correlations, the data were divided into two sets: (1) subjects with RT shorter than M2(offset) (fast group) and (2) subjects with RT more than 10 ms longer than their M2(offset) (slow group). Modulation during M2 period was large for the fast group, and it was almost zero for the slow group. These results indicate that the superimposition of RT activity mainly contributes to the instruction-dependent modulation of M2 peak.

Recovery of human motoneurons during rotation.

During prolonged contractions, few studies have reported rotation among low threshold motoneurons. The question arises whether a motoneuron stops firing due to an increase in firing threshold or whether it is due to regional switching of activity among muscle fascicles. We postulated that if the rest period resulted from an increase in firing threshold, a progressive recovery in the excitability of the motoneuron would be observed during the rest period. The excitability of soleus or tibialis anterior motoneurons was tested during the rest periods. The results showed that a previously tonic motoneuron that had dropped off during rotation, rarely responded to Ia or TMS inputs in the initial parts of the rest period; however, its response probability increased significantly in the second half. Based on these data, we suggest that the observed rotation is due to changes in firing thresholds of motoneurons during prolonged firing.

Bilateral responses of upper limb muscles to transcranial magnetic stimulation in human subjects.

Anatomical and behavioural work on primates has shown bilateral innervation of axial and proximal limb muscles, and contralateral control of distal limb muscles. The following study examined if a clear boundary exists between the distal and proximal upper limb muscles that are controlled contralaterally or bilaterally. The right motor cortical area representing the upper limb was stimulated, while surface EMG was recorded bilaterally from various upper limb muscles during rest and phasic voluntary contractions. Peak-to-peak amplitude of motor evoked potential (MEP) was measured for each muscle on both sides. The ratio R = (ipsilateral MEP: contralateral MEP) was calculated for seven pairs of muscles. For each of the seven pairs, R was less than 1.0, implying that for each muscle and subject, the contralateral control is stronger. The boundary where R changed from almost zero to a clearly measurable magnitude depended on the subject. Ipsilateral MEPs from trapezius and pectoralis could be recorded with a small background contraction from almost all subjects; on the other hand, in deltoid and biceps brachii, ipsilateral MEPs were observed only with bimanual phasic contractions. The forearm and hand muscles, in general, did not show any ipsilateral MEPs. Major differences between subjects lay in the presence or the absence of ipsilateral MEPs in biceps brachii and deltoid, without defining a sharp boundary between proximal and distal muscles.

Properties of human motor units after prolonged activity at a constant firing rate.

The primary purpose of this study was to examine if there are changes in the intrinsic properties of spinal motoneurons after prolonged submaximal contractions. To do this, we assessed whether or not the synaptic drive to motoneurons needs to increase in order to maintain a constant firing rate of a motor unit. Recruitment of new units and an increase in total electromyographic (EMG) activity of the muscle of interest were taken as estimates of an increase in synaptic drive. Subjects were asked to maintain a constant firing rate of a clearly identifiable (targeted) motor unit from the first dorsal interosseous muscle for approximately 10 min, while surface EMG and force were recorded simultaneously. For the 60 units studied, the duration of the constant-firing-rate period ranged from 73 to 1,140 s (448 +/- 227 s; mean +/- SD). There was a significant increase ( t-test, p<0.001) in the magnitude of mean surface EMG, and DC force while the targeted motoneuron maintained a constant rate suggesting an increase in the net excitatory input to the motoneuron pool. Changes occurring simultaneously in other parameters, namely, variability in interspike interval, magnitude of force fluctuations, the duration of motor unit action potentials, and the median power frequency of surface EMG were also computed. The firing rates of 16 concurrently firing motoneurons, not controlled by the subject, remained constant. The key finding of this study is that after prolonged activity, a motoneuron requires a stronger excitatory input to maintain its firing rate. Additional results are indicative of significant changes in the characteristics of the synaptic inputs, changes at the neuromuscular junction (both pre- and postsynaptic regions) and the sarcolemma.

Oxygen saturation as a screening test for critical congenital heart disease: a preliminary study.

The aim of this study was to assess the utility of arm and leg oxygen saturation as a candidate screening test for the early detection of ductal-dependent left heart obstructive disease. We measured arm and leg oxygen saturation in 2876 newborns admitted to well baby nurseries and 32 newborns with congenital heart disease. Fifty-seven newborns in the well baby nurseries (0.02%) had an abnormal test (leg saturation less than 92% in room air or 7% lower saturation in the leg than in the arm). Four of the 57 had critical congenital heart disease, including 1 with coarctation of the aorta. Of the 32 newborns with congenital heart disease, 11/13 (85%) with left heart obstructive disease had abnormal oxygen saturation tests, as did 15/19 (79%) with other forms of congenital heart disease. Pulse oximetry deserves further study as a screening test for critical congenital heart disease.

Responses of ankle extensor and flexor motoneurons to transcranial magnetic stimulation.

Transcranial magnetic stimulation (TMS) of the motor cortex excites limb muscles of the contralateral side of the body. Reports of poorly defined, or a complete lack of systematic excitatory responses of soleus motoneurons compared with those of tibialis anterior (TA) motoneurons has led to the proposal that while all ankle flexor motoneurons receive strong corticomotoneuronal connections, very few soleus motoneurons do. In addition, the connections to these few motoneurons are weak. The nature of corticomotoneuronal connections onto these two motoneuron pools was re-evaluated in the following experiments. The leg area of the left motor cortex was stimulated with a large double-cone coil using Magstim 200, while surface electromyographic (EMG) and single motor unit (SMU) responses were recorded from soleus and TA muscles of healthy adult subjects. Under resting conditions, the onset (25-30 ms) and duration of concomitantly recorded short latency motor evoked potentials (MEPs) in surface EMG from both muscles were similar. The input-output relationships of the simultaneously recorded soleus and TA EMG responses showed much greater increases in TA MEPs compared with soleus MEPs with identical increases in stimulus intensity. Under resting and nonisometric conditions, a later peak with onset latency of approximately 100 ms was observed in soleus. During isometric conditions or with vibration of the TA tendon, the second soleus peak was abolished indicating reflex origin of this peak. Recordings from 42 soleus and 39 TA motor units showed clear response peaks in the peristimulus time histograms (PSTHs) of every unit. Two statistical tests were done to determine the onset and duration of response peaks in the PSTHs. With chi(2) test, the duration was 6.9 +/- 4.2 ms (mean +/- SD) for soleus and 5.1 +/- 2.1 ms for TA. Using the criterion of discerning a peak by bin counts being three SDs above background, the duration was 10.0 +/- 4.4 ms for soleus and 7.8 +/- 2.6 ms for TA. Results of these experiments do not suggest a lack of systematic corticomotoneuronal connections on soleus motoneurons when compared with those on TA, though some differences in the strengths of corticomotoneuronal connections onto the two pools do exist.

Suicidal behavior and parental psychopathology in hospitalized depressed children.

This study describes the suicidal behavior of hospitalized depressed children and assesses its relationship to psychopathology and suicidal behavior in their parents. Subjects were 58 consecutively hospitalized prepubertal children with a primary diagnosis of major depressive disorder (MDD), and 58 age- and gender-matched children hospitalized for psychiatric diagnoses other than a mood disorder. Clinical interviews and structured diagnostic instruments were reviewed to determine the children's suicidal behavior and their parents' history of psychopathology. Suicidal ideation, suicidal intent, suicidal plans, and suicide attempts were more frequent in MDD children compared to nondepressed children. When MDD and control samples were stratified as to presence of suicidal behavior in the child, psychopathology was high in parents from all subgroups. Intensity of suicidal behavior in the depressed and non-depressed children was not associated with an altered pattern of psychopathology in their parents. Hospitalized MDD children had increased suicidal behavior compared to inpatient psychiatric control children. However, suicidal behavior in the children was not associated with increased psychopathology or an altered pattern of psychopathology in their parents.

Motor unit recruitment during lengthening contractions of human wrist flexors.

The purpose of this study was to revisit the question of recruitment of motor units during lengthening contractions because of conflicting views in the literature on this subject. Motor unit activity was recorded from the flexor carpi radialis muscle of four human subjects to compare the patterns of recruitment during lengthening and isometric contractions. Lengthening contractions were produced either when the subject voluntarily stopped opposing a background load or when an additional load was imposed on the already contracting muscle. In both cases, lengthening of the active muscle was produced at a variety of speeds, from quite slow to "as fast as possible." No differences in recruitment order were observed between isometric and lengthening contractions at any speed of lengthening contraction. It is concluded that all contractions in normal humans recruit motor units in an orderly fashion from small to large, according to the size principle of motor unit recruitment.

Control of the wrist joint in humans.

As one considers changes in motor activity from lower mammals to higher primates, one of the major changes one observes lies in the cortical control of forelimb muscles. There has been a shift from disynaptic control of spinal motoneurons in, for example, the cat, to a greater and greater percentage of monosynaptic control of hand and forelimb motoneurons in the primate. In spite of the species and evolutionary changes in the synaptic connections of the corticospinal tract, it appears that the interneurons identified in the cat are retained in the monkey and human. These interneurons, under the influence of descending pathways, modulate the output of motoneuron pools. Perhaps the control of these interneurons has also changed towards finer control of movement, as has been suggested by recent studies in the monkey. Whether in cat or human, the recruitment pattern for motor units is the same; the change from disynaptic to monosynaptic connections has not changed the recruitment pattern of muscles. Differences in the recruitment patterns of muscles may lie in the finer control of inputs to motoneurons in the primate. This review seeks to integrate the current knowledge of the mechanisms involved in the motor control of the wrist joint and especially in the recruitment patterns of the muscles. These motor control mechanisms include the biomechanics of the wrist joint, recruitment patterns of wrist muscles, interneurons and spinal cord circuits in the cervical regions mediating the output of spinal motoneurons, and the supraspinal control of these muscles.

Reduced short and long latency reflexes during voluntary tracking movement of the human wrist joint.

In six healthy human subjects we compared changes in the strength of Hoffmann (H), short latency (30-55 ms) and long latency (55-100 ms) stretch reflexes of flexor carpi radialis (FCR) muscle during movement and isometric contractions. In one set of experiments, stretches were imposed to the wrist during voluntarily tracked sinusoidal movement and during matched isometric contractions to compare short and long latency stretch reflex responses. In the second set, H-reflexes were compared during similar matched conditions. All reflexes decreased significantly (P < 0.05) during the voluntary tracking movement. The H-reflex was reduced during the wrist flexion, on average, by 33% of its value obtained during the isometric condition. Compared with their values during isometric conditions, the short latency stretch reflex and long latency stretch reflex during movement were reduced by 52 and 40%, respectively. From the pattern changes of the stretch reflexes and the H-reflex, a movement-induced presynaptic inhibition combined with pronounced muscle spindle unloading is proposed to play an important role in decreasing the strength of the stretch reflexes during the tracking task as compared with a matched isometric contraction.

Comparison of the depression of H-reflexes following previous activation in upper and lower limb muscles in human subjects.

When conditioning-testing (C-T) stimuli are applied to Ia afferents to elicit H-reflexes, the test reflex is abolished immediately following the conditioning reflex. As the C-T interval is increased, the test response slowly begins to recover, taking several hundred milliseconds to attain control values. The time course of this recovery is known as the H-reflex recovery curve. H-reflex recovery curves were compared using surface EMG and single motor unit activities in lower limb soleus and upper limb flexor carpi radialis (FCR) muscles in seven healthy human subjects. Under rest conditions, the recovery of H-reflexes and single motor unit activity was slow for soleus; the recovery was not complete even in 1 s. In comparison, the recovery was very fast for FCR motor units, occurring in 200-300 ms. The effects of rate of stimulation (0.1-10.0 imp/s) were also examined on the magnitude of H-reflex responses. The reflex response declined with increasing rate of stimulation, the decline being slightly greater in soleus than in FCR. When these phenomena were examined with voluntary facilitation of the spinal cord, the time of recovery shortened and the effect of stimulus rate also diminished. Changes with background facilitation were greater in FCR than in soleus. The differences between the two muscles are attributed mainly to differences in presynaptic inhibition in the two spinal segments, and/or to the differences in dynamics of the transmitter release in terminals of Ia afferents synapsing with slow soleus motoneurons and those synapsing with the fast FCR motoneurons.

A comparison of human motoneuron data to simulated data using cat motoneuron models.

The response of repetitively firing human motoneurons to a composite excitatory input was evaluated. It was clearly shown that the response of the motoneurons to the transient input decreased with an increase in the background firing rate of the cell. The current model of repetitively firing human motoneurons could not account for this experimental result. Therefore, a compartmental modelling approach was used to simulate the repetitive firing properties of anaesthetised cat motoneurons under current clamp conditions. The modelled motoneurons were used in simulations similar to the experimental paradigms where the response to a composite excitatory input was evaluated at different background firing rates. The motoneuron models also showed a decrease in response to the excitatory input at faster background firing rates. The results suggest that human motoneurons are more comparable to motoneurons in the anaesthetised cat preparation than formerly thought. The results also demonstrate that the apparent efficacy of a synaptic input may be modulated by changes in background firing rate of the postsynaptic neuron.

Synaptic connections from large afferents of wrist flexor and extensor muscles to synergistic motoneurones in man.

Short-latency excitatory Ia reflex connections were determined between pairs of human wrist flexor and extensor muscles. Spindle Ia afferents were stimulated by either tendon tap or electrical stimulation. The activity of voluntarily activated single motor units was recorded intramuscularly from pairs of wrist flexor or extensor muscles. Cross-correlation between stimuli and the discharge of the motor units provided a measure of the homonymous or heteronymous excitatory input to a motoneurone. Homonymous motoneurone facilitation was generally stronger than that of the heteronymous motoneurones. The principal wrist flexors, flexor carpi radialis (FCR) and flexor carpi ulnaris (FCU), were tightly connected through a bidirectional short-latency reflex pathway. In contrast, the extensor carpi ulnaris (ECU) and the extensor carpi radialis (ECR) did not have similar connections. ECU motoneurones received no short-latency excitatory Ia input from the ECR. ECR motoneurones did receive excitatory Ia input from ECU Ia afferents; however, its latency was delayed by several milliseconds compared with other heteronymous Ia excitatory effects observed. The wrist and finger extensors were linked through heteronymous Ia excitatory reflexes. The reflex connections observed in humans are largely similar to those observed in the cat, with the exception of heteronymous effects from the ECU to the ECR and from the extensor digitorum communis (EDC) to the ECU, which are present only in humans. The differences in the reflex organization of the wrist flexors versus the extensors probably reflects the importance of grasping.

Omeprazole for severe reflux esophagitis in children.

BACKGROUND: Severe esophagitis is a rare complication of gastroesophageal reflux in children. In adults, omeprazole therapy of severe erosive esophagitis has become the gold standard short-term treatment of the disease. In children, data on its use are limited, and problems about the dosage are unresolved. The aim of this study was to evaluate the efficacy of a simplified, body-weight-based daily dosage of omeprazole in children with severe esophagitis. METHODS: Ten children (median age 75.6 months; range 25-109 months) with severe esophagitis were prospectively investigated. All patients were evaluated by endoscopy, histology, and 24-h pH-metry study before and after 3 months of omeprazole. The starting dose of omeprazole was 20 mg as a single daily dose in children weighing less than 30 kg, and 40 mg daily for those weighing over 30 kg. RESULTS: A significant improvement in all the children was demonstrated after 3 months of treatment by clinical, endoscopic, and pH-metry assessment. However, histologic study failed to show significant improvement of both inflammatory and hyperplastic findings. Relapse occurred in six of 10 patients after discontinuation of therapy. CONCLUSIONS: Omeprazole is effective in the short-term treatment of severe oesophagitis in children. The daily dose of the drug could be easily based on the body weight. The persistence of histologic features of esophagitis in spite of clinical and endoscopic healing could be an indicator of poor outcome.

Computer simulation of the responses of human motoneurons to composite 1A EPSPS: effects of background firing rate.

Two compartmental models of spinal alpha motoneurons were constructed to explore the relationship between background firing rate and response to an excitatory input. The results of these simulations were compared with previous results obtained from human motoneurons and discussed in relation to the current model for repetitively firing human motoneurons. The morphologies and cable parameters of the models were based on two type-identified cat motoneurons previously reported in the literature. Each model included five voltage-dependent channels that were modeled using Hodgkin-Huxley formalism. These included fast Na+ and K+ channels in the initial segment and fast Na+ and K+ channels as well as a slow K+ channel in the soma compartment. The density and rate factors for the slow K+ channel were varied until the models could reproduce single spike AHP parameters for type-identified motoneurons in the cat. Excitatory synaptic conductances were distributed along the equivalent dendrites with the same density described for la synapses from muscle spindles to type-identified cat motoneurons. Simultaneous activation of all synapses on the dendrite resulted in a large compound excitatory postsynaptic potential (EPSP). Brief depolarizing pulses injected into a compartment of the equivalent dendrite resulted in pulse potentials (PPs), which resembled the compound EPSPs. The effects of compound EPSPs and PPs on firing probability of the two motoneuron models were examined during rhythmic firing. Peristimulus time histograms, constructed between the stimulus and the spikes of the model motoneuron, showed excitatory peaks whose integrated time course approximated the time course of the underlying EPSP or PP as has been shown in cat motoneurons. The excitatory peaks were quantified in terms of response probability, and the relationship between background firing rate and response probability was explored. As in real human motoneurons, the models exhibited an inverse relationship between response probability and background firing rate. The biophysical properties responsible for the relationship between response probability and firing rate included the shapes of the membrane voltage trajectories between spikes and nonlinear changes in PP amplitude during the interspike interval at different firing rates. The results from these simulations suggest that the relationship between response probability and background firing rate is an intrinsic feature of motoneurons. The similarity of the results from the models, which were based on the properties of cat motoneurons, and those from human motoneurons suggests that the biophysical properties governing rhythmic firing in human motoneurons are similar to those of the cat.

Activation of midlumbar neurones by afferents from anterior hindlimb muscles in the cat.

1. It has been suggested that a group of interneurones located in the midlumbar segments of the spinal cord might play a role in switching from the stance to swing phases of the step cycle during locomotion. We have further examined the input to these neurones from proprioceptive afferents to test whether the connections to these cells are consistent with this role. 2. Electrical stimulation of group I and group II afferents in branches of the femoral nerve which supply iliopsoas, the major hip flexor muscle, excited a large majority of intermediate zone midlumbar interneurones which receive input from quadriceps group II afferents. The central latencies and properties of the EPSPs indicate that both group I and group II afferents from iliopsoas make monosynaptic connections with many midlumbar interneurones. 3. Group II afferents from both the ankle flexor tibialis anterior and the digit dorsiflexor extensor digitorum longus excited midlumbar interneurones. Similarly, they were also excited by group II afferents from both of the two main anatomical divisions of the sartorius muscle. 4. The frequent and potent excitation of midlumbar neurones from group I and II afferents in iliopsoas suggests that they may be excited at the end of the stance phase of the step when these muscles are stretched. This possibility is discussed in relation to recent work on the functional control of the step cycle.