Energy exchange between subject and belt during treadmill walking

Treadmill walking aims to simulate overground walking, but intra-stride belt speed variations of treadmills result in some interaction between treadmill and subject, possibly obstructing this aim. Especially in self-paced treadmill walking, in which the belt speed constantly adjusts to the subject, these interactions might affect the gait pattern significantly. The aim of this study was to quantify the energy exchange between subject and treadmill, during the fixed speed (FS) and self-paced (SP) modes of treadmill walking. Eighteen subjects walked on a dual-belt instrumented treadmill at both modes. The energy exchange was calculated as the integration of the product of the belt speed deviation and the fore-aft ground reaction force over the stride cycle. The total positive energy exchange was 0.44 J/stride and the negative exchange was 0.11 J/stride, which was both less than 1.6% of the performed work on the center of mass. Energy was mainly exchanged from subject to treadmill during both the braking and propulsive phase of gait. The two treadmill modes showed a similar pattern of energy exchange, with a slightly increased energy exchange during the braking phase of SP walking. It is concluded that treadmill walking is only mildly disturbed by subject-belt interactions when using instrumented treadmills with adequate belt control.     

Can treadmill walking be used to assess propulsion generation?

Instrumented treadmills offer significant advantages for analysis of human locomotion, including recording consecutive steady-state gait cycles, precisely controlling walking speed, and avoiding force plate targeting. However, some studies of hemiparetic walking on a treadmill have suggested that the moving treadmill belt may fundamentally alter propulsion mechanics. Any differences in propulsion mechanics during treadmill walking would be problematic since recent studies assessing propulsion have provided fundamental insight into hemiparetic walking. The purpose of this study was to test the hypothesis that there would be no difference in the generation of anterior/posterior (A/P) propulsion by performing a carefully controlled comparison of the A/P ground reaction forces (GRFs) and impulses in healthy adults during treadmill and overground walking. Gait data were collected from eight subjects walking overground and on a treadmill with speed and cadence controlled. Peak negative and positive horizontal GRFs in early and late stance, respectively, were reduced by less than 5% of body weight (p<0.05) during treadmill walking compared to overground walking. The magnitude of the braking impulse was similarly lower (p<0.05) during treadmill walking, but no significant difference was found between propulsion impulses. While there were some subtle differences in A/P GRFs between overground and treadmill walking, these results suggest there is no fundamental difference in propulsion mechanics. We conclude that treadmill walking can be used to investigate propulsion generation in healthy and by implication clinical populations.     

Fractal fluctuations in spatiotemporal variables when walking on a self-paced treadmill

This study investigated the fractal dynamic properties of stride time (ST), stride length (SL) and stride speed (SS) during walking on a self-paced treadmill (STM) in which the belt speed is automatically controlled by the walking speed. Twelve healthy young subjects participated in the study. The subjects walked at their preferred walking speed under four conditions: STM, STM with a metronome (STM+met), fixed-speed (conventional) treadmill (FTM), and FTM with a metronome (FTM+met). To compare the fractal dynamics between conditions, the mean, variability, and fractal dynamics of ST, SL, and SS were compared. Moreover, the relationship among the variables was examined under each walking condition using three types of surrogates. The mean values of all variables did not differ between the two treadmills, and the variability of all variables was generally larger for STM than for FTM. The use of a metronome resulted in a decrease in variability in ST and SS for all conditions. The fractal dynamic characteristics of SS were maintained with STM, in contrast to FTM, and only the fractal dynamic characteristics of ST disappeared when using a metronome. In addition, the fractal dynamic patterns of the cross-correlated surrogate results were identical to those of all variables for the two treadmills. In terms of the fractal dynamic properties, STM walking was generally closer to overground walking than FTM walking. Although further research is needed, the present results will be useful in research on gait fractal dynamics and rehabilitation.     

Force measurements during running on different instrumented treadmills

One method to determine the forces produced during running is to conduct extensive kinematic and kinetic analysis. These analyses can be performed by having an individual perform repeated over-ground running trials or simply run continuously on an instrumented treadmill. The forces produced during over-ground running may not be the same as the forces during treadmill running and these differences could be attributed to a number of factors, including the design of the instrumented treadmill. The purpose of this paper was to determine whether there are differences in force measurements on different instrumented treadmill setups in comparison to over-ground running and to correct for any of these differences using a theoretical model. 11 participants ran on three different treadmills and performed over-ground running at 2.7, 3.6, and 4.5 m/s. Ground reaction forces were measured via force plates and an instrumented pressure insole. We found that the magnitude of the vertical ground reaction force differed between the three treadmills and over-ground running. The difference in ground reaction forces estimated by the pressure insole and the treadmill-force-plate system or instrumented treadmill can be explained by a three degree of freedom mechanical model of a person running on a treadmill and this model could potentially be used to correct for errors in force measurement from instrumented treadmills. The model included a force plate, a treadmill, and a wobbling mass with varying natural frequencies and damping characteristics, and constant masses. These findings provide researchers a method to correct forces from an instrumented treadmill set-up to determine a close approximation of the actual forces experienced by a participant during treadmill running.     

EMG activity during positive-pressure treadmill running

Success has been demonstrated in rehabilitation from certain injuries while using positive-pressure treadmills. However, certain injuries progress even with the lighter vertical loads. Our purpose was to investigate changes in muscle activation for various lower limb muscles while running on a positive-pressure treadmill at different amounts of body weight support. We hypothesized that some muscles would show decreases in activation with greater body weight support while others would not.Eleven collegiate distance runners were recruited. EMG amplitude was measured over 12 lower limb muscles. After a short warm-up, subjects ran at 100%, 80%, 60%, and 40% of their body weight for two minutes each. EMG amplitudes were recorded during the final 30 s of each stage.Most muscles demonstrated lower amplitudes as body weight was supported. For the hip adductors during the swing phase and the hamstrings during stance, no significant trend appeared.Positive-pressure treadmills may be useful interventions for certain injuries. However, some injuries, such as hip adductor and hamstring tendonitis or strains may require alternative cross-training to relieve stress on those areas. Runners should be careful in determining how much body weight should be supported for various injuries to return to normal activity in the shortest possible time.     

Local dynamic stability versus kinematic variability of continuous overground and treadmill walking

This study quantified the relationships between local dynamic stabiliht and variabilitr during continuous overground and treadmill walking. Stride-to-stride standard deviations were computed from temporal and kinematic data. Marimum finite-time Lyapunov exponents were estimated to quantify local dynamic stability. Local stability of gait kinematics was shown to be achieved over multiple consecutive strides. Traditional measures of variability poorly predicted local stability. Treadmill walking was associated with significant changes in both variability and local stability. Thus, motorized treadmills may produce misleading or erroneous results in situations where changes in neuromuscular control are likely to affect the variability and/or stability of locomotion.     

Effects of size, sex, and voluntary running speeds on costs of locomotion in lines of laboratory mice selectively bred for high wheel-running activity

Selective breeding for over 35 generations has led to four replicate (S) lines of laboratory house mice (Mus domesticus) that run voluntarily on wheels about 170% more than four random-bred control (C) lines. We tested whether S lines have evolved higher running performance by increasing running economy (i.e., decreasing energy spent per unit of distance) as a correlated response to selection, using a recently developed method that allows for nearly continuous measurements of oxygen consumption (VO2) and running speed in freely behaving animals. We estimated slope (incremental cost of transport [COT]) and intercept for regressions of power (the dependent variable, VO2/min) on speed for 49 males and 47 females, as well as their maximum VO2 and speeds during wheel running, under conditions mimicking those that these lines face during the selection protocol. For comparison, we also measured COT and maximum aerobic capacity (VO2max) during forced exercise on a motorized treadmill. As in previous studies, the increased wheel running of S lines was mainly attributable to increased average speed, with males also showing a tendency for increased time spent running. On a whole-animal basis, combined analysis of males and females indicated that COT during voluntary wheel running was significantly lower in the S lines (one-tailed P=0.015). However, mice from S lines are significantly smaller and attain higher maximum speeds on the wheels; with either body mass or maximum speed (or both) entered as a covariate, the statistical significance of the difference in COT is lost (one-tailed P> or =0.2). Thus, both body size and behavior are key components of the reduction in COT. Several statistically significant sex differences were observed, including lower COT and higher resting metabolic rate in females. In addition, maximum voluntary running speeds were negatively correlated with COT in females but not in males. Moreover, males (but not females) from the S lines exhibited significantly higher treadmill VO2max as compared to those from C lines. The sex-specific responses to selection may in part be consequences of sex differences in body mass and running style. Our results highlight how differences in size and running speed can account for lower COT in S lines and suggest that lower COT may have coadapted in response to selection for higher running distances in these lines.     

Adjustment of flight speed of gregarious desert locusts (Orthoptera: Acrididae) flying side by side

In tethered flying locusts, optomotor thrust responses induced by translatory pattern motion within the lateral visual fields were studied under closed-loop conditions. By modulating thrust in a compensatory manner, locusts counteracted a bias motion superposed on the thrust-related motion. This way, pattern speed was kept at 0° s^–1, indicating the set point of the respective optomotor control circuit. Though the quality of bias compensation varied greatly, it was largely independent from pattern characteristics. It might indicate that the gain of behavior not only is controlled by an automatic mechanism but also is affected by spontaneous modulations. Compensation of bias motion was critically dependent on the relation between self- and bias-generated motion: Locusts did not take control over pattern motion if self- and bias-generated motion differed greatly. Instead, locusts adopted a constant, supposingly preferred, thrust value. Therefore, flight speed is assumed to be controlled by two systems: the optomotor and a preferred thrust system. In free flight, an equalization of the flight speed of locusts within a swarm might result from similar behavior. In combination with a presumed coordination of the locusts' course direction, this may explain the continued cohesion of swarms in the field.     

Optimization of behavior by minimization of displeasure in a 2-dimensional sensory space

Three healthy young adult males were placed on a treadmill geared to five speeds and five slopes in combination. At minute 16 they gave estimates of displeasure for sensations aroused in their chest and lower limbs. These estimates were compared to the operant choice of the subjects when they were given the task, in other sessions, of climbing up 300 m on the treadmill at varying speeds and slopes. When speed was imposed, they could adjust slope and reciprocally. The were instructed that the session would end when they had reached the 300 m elevation. The results showed that the subjects adjusted slope and speed inversally; time for climbing up 300 m was almost constant, i.e. they tended to work at constant power in various conditions. Such performance could be predicted from the sum of ratings of displeasure aroused in the lower limbs and in the chest. These results suggest that optimalization of behaviour was obtained by minimization of bidimensional displeasure.     

Involvement of a cell size control mechanism in the induction and maintenance of oscillations in continuous cultures of budding yeast

Spontaneous oscillations occur in glucose-limited continuous cultures of Saccharomyces cerevisiae under aerobic conditions. The oscillatory behavior is detectable as a periodic change of many bioparameters such as dissolved oxygen, ethanol production, biomass concentration, as well as cellular content of storage carbohydrates and is associated to a marked synchronization of the yeast population. These oscillations may be related to a periodic accumulation of ethanol produced by yeast in the culture medium.The addition of ethanol to oscillating yeast cultures supports this hypothesis: indeed, no effect was observed if ethanol was added when already present in the medium, while a marked phase oscillation shift was obtained when ethanol was added at any other time. Moreover, the addition of ethanol to a nonoscillating culture triggers new oscillations. An accurate analysis performed at the level of nonoscillating yeast populations perturbed by addition of ethanol showed that both the growth rate and the protein content required for cell division increased in the presence of mixed substrate (i.e., ethanol plus limiting glucose). A marked synchronization of the yeast population occurred when the added ethanol was exhausted and the culture resumed growth only on limiting glucose. A decrease of protein content required for cell division was also apparent. These experimental findings support a new model for spontaneous oscillations in yeast cultures in which the alternative growth on limiting glucose and limiting glucose plus ethanol modifies the critical protein content required for cell division.     

群体成员大小差异对不同生境鲤科鱼类集群行为的影响

为研究群体成员大小差异对不同喜好生境鱼类集群行为特征的影响, 实验分别以鳊(Parabramis pekinensis)和中华倒刺鲃(Spinibarbus sinensis)幼鱼为实验对象, 比较分析4尾等大小(E)和不等大小(2大2小, NE)实验鱼群体的自发游泳速度、空间分布以及对恐吓刺激反应等集群行为参数的差异。结果显示: (1)和鳊相比, 中华倒刺鲃有更高的自发游泳速度、速度同步性和排列方向的极性, 但二者对恐吓刺激的反应率及反应的协调一致性相似; (2)当群体成员大小出现差异时, 两种鱼群体排列方向的极性不受影响, 且大小个体成员间的速度及其同步性均没有差异, 但整体的速度同步性与等大小群体相比有所下降; (3)个体间距离数据显示, 个体大小差异不会影响两种鱼群体的凝聚力; (4)群体成员在两种鱼群中偏好位置不同, 当群体成员大小不同时, 大个体成员更偏好占据领头鱼位置; (5)群体成员大小的差异导致两种鱼对刺激的反应率下降。研究表明: 中华倒刺鲃具有更高的活跃性、更好的群体运动的协调性, 可能与其流水生境相关; 当群体成员大小出现差异时, 成员不分大小在整体上协调运动的速度和方向, 并保持群体有较高的凝聚力, 但两种鱼类自发游泳速度调整策略截然不同(鳊大小个体速度妥协趋同, 而中华倒刺鲃低速个体速度提高); 群体成员大小差异导致鱼群对恐吓刺激的反应率有所下降, 可能原因包括体形差异导致的社会因素造成敏锐性下降、信息交流效率受阻和(或)集群收益代价出现分化影响一致决策的形成等。     

Metabolism of rats running up and down an incline

The purpose of these experiments was to determine oxygen consumption (VO2) in rats as a function of treadmill speed (10, 20, 30, 40, and 50 m . min-1) as they ran on the level and up and down a 16 degree (17.8%) incline. The slopes of the regression lines relating VO2 (ml O2 . kg-1 . min-1) to running speed (m . min-1) were linear for all three inclines. The regression slope for uphill runners (y = 1.25x + 47.7) was greater than the regression slopes for level (y = 0.88x + 41.2) (P less than 0.025) or downhill (y = 0.68x + 39.7) (P less than 0.005) runners, and the regression slope for level runners was greater than that for downhill runners (P less than 0.10). All VO2 measurements were submaximal. In conclusion, incline has a significant effect on the metabolism of rats running on a motor-driven treadmill.     

A comparison of the oxygen consumption/body weight relationship obtained during submaximal exercise on a bicycle ergometer and on a treadmill.

It is widely accepted that the relationship between oxygen consumption and body weight obtained during exercise on a bicycle ergometer differs from that obtained during treadmill walking. Experimental evidence to support this claim is lacking. To examine this difference a group of subjects (body weight 41--81 kg) undertook a predetermined level of submaximal exercise on a bicycle ergometer and a treadmill. Oxygen consumption was measured in a steady state at rest (i.e. sitting on the bicycle ergometer and standing on the treadmill) and during the two modes of exercise. A significant positive correlation between oxygen consumption and body weight was obtained under all four conditions of measurement. At rest the two regression lines did not differ in slope or elevation. During exercise the slope and the elevation of the line obtain from treadmill walking were significantly greater than from bicycle ergometer exercise. The 'metabolic cost' of bicycle ergometer exercise, (Vo2 during exercise--V02 at rest), showed no significant correlation with body weight. In contrast, there was a significant positive correlation during walking. It is suggested that these differences have arisen due to a different proportion of the total body weight supported by the subject in the two forms of exercise.     

Improving LC-MS sensitivity through increases in chromatographic performance: comparisons of UPLC-ES/MS/MS to HPLC-ES/MS/MS

Recent technological advances have made available reverse phase chromatographic media with a 1.7 microm particle size along with a liquid handling system that can operate such columns at much higher pressures. This technology, termed ultra performance liquid chromatography (UPLC), offers significant theoretical advantages in resolution, speed, and sensitivity for analytical determinations, particularly when coupled with mass spectrometers capable of high-speed acquisitions. This paper explores the differences in LC-MS performance by conducting a side-by-side comparison of UPLC for several methods previously optimized for HPLC-based separation and quantification of multiple analytes with maximum throughput. In general, UPLC produced significant improvements in method sensitivity, speed, and resolution. Sensitivity increases with UPLC, which were found to be analyte-dependent, were as large as 10-fold and improvements in method speed were as large as 5-fold under conditions of comparable peak separations. Improvements in chromatographic resolution with UPLC were apparent from generally narrower peak widths and from a separation of diastereomers not possible using HPLC. Overall, the improvements in LC-MS method sensitivity, speed, and resolution provided by UPLC show that further advances can be made in analytical methodology to add significant value to hypothesis-driven research.     

Autophagic response to a single bout of moderate exercise in murine skeletal muscle

The effect of a single bout of exercise on autopahgy in murine gastrocnemius muscle was investigated. Autophagy is a process for the degradation system of cytoplasmic components, which may help maintain intracellular quality control of cell survival and turnover under normal conditions. The present study investigated the changes of autophagy-related proteins including microtubule-associated protein 1b light chain 3 (LC3), Beclin-1, Atg7 (autophagy-related gene 7), conjugation form of Atg12 to Atg5, lysosome-associated membrane protein (LAMP2a), and muscle-specific RING finger protein-1 (MURF-1) protein level in gastrocnemius muscle after a single bout of treadmill exercise. Mice exercised on a treadmill for 50 min at a speed of 12.3 m/min with a slope of 5°. The animals were sacrificed by cervical dislocation 0, 3, 6, or 12 h after exercise, and muscle samples were collected immediately. Western blot analysis demonstrated that the autophagy marker LC3-II was significantly decreased during the recovery period (3, 6, and 12 h) whereas there was no decrease immediately after exercise (0 h). To identify factors related to this decrease, autophagosome component proteins were examined in murine gastrocnemius muscle. A decrease in Beclin-1, Atg7, and LAMP2a during recovery period was concomitant with the decreased level of LC3-II. Additionally, MuRF-1 expression was significantly increased after a single bout of exercise. This study is the first to demonstrate autophasic related protein expression after a single bout of treadmill exercise and our results suggest that a single bout of treadmill exercise attenuates the autophagic response in murine skeletal muscle.     

BEHAVIOR OF RED-TAILED HAWKS IN A WIND TURBINE DEVELOPMENT

Abstract: Birds flying within windfarms can be killed when they collide with wind turbines. Raptors, especially red-tailed hawks (Buteo jamaicensis), are more susceptible to collisions than other birds, which may be attributable to their specific foraging and flight behavior. To more fully understand the problem, and to reduce raptor mortality, it is necessary to acquire more information on habitat use and flight behavior by raptors inhabiting windfarms. Between June 1999 and June 2000, we watched raptors for 346 hours in the Altamont Pass Wind Resource Area, the largest windfarm in North America. We recorded flight behavior in relation to characteristics of the topography such as slope aspect, elevation, and inclination and in relation to various weather variables including wind speed and wind direction. We found that red-tailed hawk behavior and their use of slope aspect differed according to wind speed. Hawks perched more often in weak winds than in strong. Red-tailed hawks were more likely to soar during low wind conditions and kite during strong wind, particularly on hillsides that faced into the wind as opposed to hillsides shielded from the wind. This is likely a result of their use of deflection updrafts for lift during flight. During our study, when winds were strong and from the south-southwest, kiting behavior occurred on south-southwestern facing slopes with inclines of greater than 20% and peak elevations greater than adjacent slopes. Accordingly, mitigation measures to decrease red-tailed hawk fatalities should be directed specifically to these areas and others fitting this general model. Wind farm managers can power down turbines at the top of these hazardous slopes when they pose the greatest danger—when winds are strong and facing perpendicularly to the slope.     

Inertial compensation for belt acceleration in an instrumented treadmill

Instrumented treadmills provide a convenient means for applying horizontal perturbations during gait or standing. However, varying the treadmill belt speed introduces inertial artifacts in the sagittal plane moment component of the ground reaction force. Here we present a compensation method based on a second-order dynamic model that predicts inertial pitch moment from belt acceleration. The method was tested experimentally on an unloaded treadmill at a slow belt speed with small random variations (1.20±0.10 m/s) and at a faster belt speed with large random variations (2.00±0.50 m/s). Inertial artifacts of up to 12 Nm (root-mean-square, RMS) and 30 Nm (peak) were observed. Coefficients of the model were calibrated on one trial and then used to predict and compensate the pitch moment of another trial with different random variations. Coefficients of determination (R²$R^2$) were 72.08% and 96.75% for the slow and fast conditions, respectively. After compensation, the root-mean-square (RMS) of the inertial artifact was reduced by 47.37% for the slow speed and 81.98% for fast speed, leaving only 1.5 Nm and 2.1 Nm of artifact uncorrected, respectively. It was concluded that the compensation technique reduced inertial errors substantially, thereby improving the accuracy in joint moment calculations on an instrumented treadmill with varying belt speed.     

Synchronizing a multicellular system by external input: an artificial control strategy

MOTIVATION: Although there are significant advances on elucidating the collective behaviors on biological organisms in recent years, the essential mechanisms by which the collective rhythms arise remain to be fully understood, and further how to synchronize multicellular networks by artificial control strategy has not yet been well explored. RESULTS: A control strategy is developed to synchronize gene regulatory networks in a multicellular system when spontaneous synchronization cannot be achieved. We first construct an impulsive control system to model the process of periodically injecting coupling substances with constant or random impulsive control amounts into the common extracellular medium, and further study its effects on the dynamics of individual cells. We derive the threshold of synchronization induced by the periodic substance input. Therefore, we can synchronize the multicellular network to a specific collective behavior by changing the frequency and amplitude of the periodic stimuli. Moreover, a two-stage scheme is proposed to facilitate the synchronization in this paper. We show that the presence of the external input may also initiate different dynamics. The multicellular network of coupled repressilators is used to show the effectiveness of the proposed method. The results not only provide a perspective to understand the interactions between external stimuli and intrinsic physiological rhythms, but also may lead to development of realistic artificial control strategy and medical therapy. AVAILABILITY: CONTACT: aihara@sat.t.u-tokyo.ac.jp.     

Walking speed and spatiotemporal step mean measures are reliable during feedback-controlled treadmill walking; however,spatiotemporal step variability is not reliable

The purpose of the study was to compare the effects of a feedback-controlled treadmill (FeedbackTM) to a traditional fixed-speed treadmill (FixedTM) on spatiotemporal gait means, variability, and dynamics. The study also examined inter-session reliability when using the FeedbackTM. Ten young adults walked on the FeedbackTM for a 5-minute familiarization followed by a 16-minute experimental trial. They returned within one week and completed a 5-minute familiarization followed by a 16-minute experimental trial each for FeedbackTM and FixedTM conditions. Mean walking speed and step time, length, width, and speed means and coefficient of variation were calculated from all experimental conditions. Step time, length, width, and speed gait dynamics were analyzed using detrended fluctuation analysis. Mean differences between experimental trials were determined using ANOVAs and reliability between FeedbackTM sessions was determined by intraclass correlation coefficient. No difference was found in mean walking speed nor spatiotemporal variables, with the exception of step width, between the experimental trials. All mean spatiotemporal variables demonstrated good to excellent reliability between sessions, while coefficient of variation was not reliable. Gait dynamics of step time, length, width, and speed were significantly more persistent during the FeedbackTM condition compared to FixedTM, especially step speed. However, gait dynamics demonstrated fair to poor reliability between FeedbackTM sessions. When walking on the FeedbackTM, users maintain a consistent set point, yet the gait dynamics around the mean are different when compared to walking on a FixedTM. In addition, spatiotemporal gait dynamics and variability may not be consistent across separate days when using the FeedbackTM.     

Oligonucleotide--minor groove binder 1:2 conjugates: side by side parallel minor groove binder motif in stabilization of DNA duplex

Synthetic polycarboxamides consisting of N-methylpyrrole (Py), N-methylimidazole (Im), N-methyl-3-hydroxypyrrole (Hp) and beta-alanine (beta) show strong and sequence-specific interaction with the DNA minor groove when they form hairpin structures with side-by-side antiparallel motifs. In the present paper, new conjugates containing two ligands linked to the same terminal phosphate of DNA strand were constructed. The paper describes optimized synthesis and properties of oligonucleotide-linked polyamide strands that insert into the minor groove of a duplex in a parallel or antiparallel orientation. Strong stabilization of DNA duplexes by two attached minor groove ligands is demonstrated by the thermal denaturation method. The unmodified duplex 5'-CGTTTATTp-3'/5'-AATAAACG-3' melts at 20 degrees C. When one tetra(Py) residue was attached to the first strand of this duplex, denaturation temperature was increased to 46 degrees C; attachment of the second tetra(Py) in a parallel orientation resulted in denaturation temperature of 60 degrees C. It is even higher than in case of "classic" octapyrrole hairpin ligand (Tm = 58 degrees C). Sequence-specific character of stabilization by two conjugated ligands was demonstrated for G:C-containing oligonucleotides attached to tetracarboxamide and octacarboxamide ligands constructed from Py, Im and beta units according to established recognition rules (deltaTm = 20 degrees C). The two-strand parallel minor groove binder constructions attached to addressing oligonucleotides could be considered as site-specific ligands recognizing single- and double-stranded DNA similarly to already described hairpin MGB structures with antiparallel orientation of carboxamide units.