Internal consistency and test–retest reliability of an instrumented functional reaching task using wireless electromyographic sensors

The purpose of this study was to establish the internal consistency and test–retest reliability of the electromyographic and accelerometric data sampled from the prime movers of the dominant arm during an antigravity, within-arm’s length stand-reaching task without trunk restraint. Ten healthy young adults participated in two experimental sessions, approximately 7–10 days apart. During each session, subjects performed 15 trials of both a flexion- and an abduction-reaching task. Surface EMG and acceleration using wireless sensors were sampled from the anterior and middle deltoid. Reliability was established using Cronbach’s alpha, intraclass correlation coefficients (ICC 2, k) and standard error of measurements (SEM) for electromyographic reaction time, burst duration and normalized amplitude along with peak acceleration. Results indicated high degrees of inter-trial and test–retest reliability for flexion (Cronbach’s α range = 0.92–0.99; ICC range = 0.82–0.92) as well as abduction (Cronbach’s α range = 0.94–0.99; ICC range = 0.81–0.94) reaching. The SEM associated with response variables for flexion and abduction ranged from 1.55–3.26% and 3.33–3.95% of means, respectively. Findings from this study revealed that electromyographic and accelerometric data collected from prime movers of the arm during the relatively functional stand-reaching task were highly reproducible. Given its high reliability and portability, the proposed test could have applications in clinical and laboratory settings to quantify upper limb function.     

Convergent and construct validity and test–retest reliability of the Caen Chronotype Questionnaire in six languages

ABSTRACT

Chronotype questionnaires provide a simple and time-effective approach to assessing individual differences in circadian variations. Chronotype questionnaires traditionally focused on one dimension of chronotype, namely its orientation along a continuum of morningness and eveningness. The Caen Chronotype Questionnaire (CCQ) was developed to assess an additional dimension of chronotype that captures the extent to which individual functioning varies during the day (amplitude). The aim of this study was to provide a multilanguage validation of the CCQ in six world regions (Arabic, Dutch, German, Italian, Portuguese and Spanish). At Time 1, a total of 2788 participants agreed to take part in the study (Arabic, n = 731; Dutch, n = 538; German, n = 329; Italian, n = 473; Portuguese, n = 361; Spanish, n = 356). Participants completed an assessment of the CCQ together with the Morningness-Eveningness Questionnaire (MEQ; Horne & Ostberg 1976) as well as questions related to factors theoretically related to chronotype (age, shift work, physical activity, sleep parameters and coffee consumption). One month later, participants again completed the CCQ. Results showed that the two-factor structure (morningness-eveningness and amplitude) of the CCQ could be replicated in all six languages. However, measurement invariance could not be assumed regarding the factor loadings across languages, meaning that items loaded more on their factors in some translations than in others. Test–retest reliability of the CCQ ranged from unacceptable (German version) to excellent (Dutch, Portuguese). Convergent validity was established through small–medium effect size correlations between the morningness-eveningness dimension of the CCQ and the MEQ. Taken together, our findings generally support the use of the translated versions of the CCQ. Further validation work on the CCQ is required including convergent validation against physiological markers of sleep, health and well-being.     

The use of electromyography for the assessment of sense of muscular effort: a test–retest reliability study

This study sought to examine the test–retest reliability to measure sense of muscular effort with electromyography (EMG). The EMG activity of the tibialis anterior muscle from 23 participants was recorded. Targets of EMG amplitudes produced at 10 and 20% of the maximum voluntary contraction (MVC) were calculated. Participants matched the target EMG level with and without visual feedback (FB). With NFB, the reliability was good to excellent when errors were represented as the average standard deviation (SD) of the error from the target (ICC_1,2 = 0.75 and 0.69 for 10 and 20% targets, respectively). Also, reliability was good when errors were presented as the average SD as a percentage of the MVC EMG (intraclass correlation coefficient (ICC_1,2) = 0.67 and 0.66, respectively, for 10 and 20% targets). Standard deviation around the target was the most reliable method to represent the error. This approach could be used as a simple cost-effective method to assess the sense of muscular effort.     

Evaluation of on-farm methods for testing the human–animal relationship in dairy herds with cubicle loose housing systems—test–retest and inter-observer reliability and consistency to familiarity of test person

The human–animal relationship is an important factor when considering animal welfare at herd level. In the present study, two behavioural tests for the on-farm assessment of the human–animal relationship at herd level of dairy cows housed in loose housing cubicle systems were evaluated with respect to inter-observer reliability, test–retest reliability, effect of familiarity of test person as well as inter-correlation of the two tests. In a voluntary animal approach (VAA) test, the number of cows and the latencies to approach and touch a stationary test person was measured. In an avoidance (AV) test, the cows’ avoidance reactions to an approaching test person were categorised. A first study was carried out in 12 commercial Danish and Austrian dairy farms. On each farm, both behaviour tests were carried out on the same day and repeated within 4–5 days. For each test, cows were tested by both an unfamiliar and a familiar test person (the stock-person) and two observers simultaneously registered the animals’ test responses. The inter-observer reliability of both behavioural tests was found to be high (VAA: 2.5-m approach r=0.98 (P<0.001) and touch r=0.97 (P<0.001); AV: Kappa coefficient_weighted=0.886 (prevalence index for flight distance≥2 m is 0.636)). The cows at herd level showed shortest latency for touching an unfamiliar test person on the first test day (P=0.006). Further, the AV test had a high test–retest reliability (Kappa coefficient_weighted=0.503 (prevalence index for flight distance≥2 m is −0.660)) and results indicated no effect of familiarity of test person (Kappa coefficient_weighted=0.463 (prevalence index for flight distance≥2 m is −0.677)). In a second study, the correlation between the two behavioural tests (similar measures) was evaluated. On each of 10 commercial Danish dairy farms with loose housing cubicle systems at four repeated sessions, both behaviour tests were carried out on the same day. For each test cows were tested by the stock-person. The VAA and AV tests at herd level were highly correlated (r_s=−0.84; P=0.002).The results suggest that the AV test is valid and applicable for on-farm assessment of the human–animal relationship at herd level. This accounts only partly for the VAA test, which seem to be more unclear regarding motivation for the animals’ approach behaviour.     

Test–retest reliability for assessment of postural stability using center of pressure spatial patterns of three-dimensional statokinesigrams in young health participants

Introduction

This study investigated the test–retest reliability for assessment of postural stability using a quantitative method for identification of center of pressure (CoP) spatial patterns of three-dimensional statokinesigrams (3D-SKG).

Methods

Twenty-one healthy participants (11 women, age 26.8±7.2 years, body mass index 25.6±5.3 kg/m²) were submitted to four consecutive 60-s trials while performing undisturbed upright stance with feet together, with or without visual input each. CoP data was used to calculate parameters from the 3D-SKG (quantity of high-density regions, nHDR). Stabilogram (standard deviation, range, maximum velocity) and statokinesigram (elliptical area, average velocity) were also calculated. Intraclass correlation coefficients (ICC_2,1 and ICC_2,4) and repeated-measures analysis-of-variance were used for statistical analysis.

Results

Significant differences in nHDR among trials were not noticed in both protocols, as well as for any parameter of the stabilogram or statokinesigram (all P>0.05). Reliability for identification of nHDR with or without visual input was either excellent (ICC_2,4=0.844 and 0.792, respectively) or fair to good (ICC_2,1=0.575 and 0.488, respectively). Reliability of parameters from stabilogram and statokinesigram varied from excellent to poor for either postural task with (ICC_2,4 range: 0.961–0.491; ICC_2,1 range: 0.859–0.194) or without visual input (ICC_2,4 range: 0.990–0.444; ICC_2,1 range: 0.960–0.166).

Conclusions

Test–retest reliability for identification of CoP spatial patterns is excellent or fair to good using averaged or single measurements of nHDR, respectively. No learning effect on repeated trials for identification of CoP spatial patterns was detected but deserves further research.     

Test–retest reliability of the soleus H-reflex is affected by joint positions and muscle force levels

The purpose of this study was to determine the test–retest reliability of the soleus (SOL) H-reflex during rest and isometric contractions at 10%, 30%, and 50% of the maximal voluntary force (MVC) at the ankle joint angles of neutral (0°), plantarflexion (20°), and dorsiflexion (?20°) respectively, in a sitting position. Ten healthy participants, with mean age of 24.9 ± 5.0 (SD) years, height 168.3 ± 8.8 cm, weight 62.7 ± 12.3 kg, were tested for the SOL H-reflex (H_max) on two separate occasions within 7 days. The intraclass correlation coefficient (ICC) for the test–retest of the SOL H-reflex during rest was found to be high at ankle joint angle of neutral (ICC = 0.92) and plantarflexion (0.96), and moderate at dorsiflexion (0.75). Inconsistent ICC values (range from 0.62 to 0.97) were found during the submaximal voluntary contractions at the three ankle joint positions. High ICCs were also found in H_max/M_max ratio at neutral (0.86), plantarflexion (0.96), and dorsiflexion (0.84) positions. It was concluded that the test–retest reliability of the SOL H-reflex was affected by the intensity of voluntary contraction and ankle joint position. The H-reflex demonstrated a higher reliability at the neutral and plantarflexion positions than that at the dorsiflexion position during rest, and a higher reliability at 10% MVC than that at 30% and 50% MVC.     

Test–retest reliability of subjective supra-threshold scaling of multiple pressure-pain sensations among healthy individuals: a study using hydraulic pressure algometry

Abstract

Background: Supra-threshold scaling of multiple pressure-pain sensations involves delivery of varied stimulus intensities, either via stimulus-dependent or response-dependent manner, and recording of subjective pain ratings by participants. The focus of this study was to determine the intra- and inter-session reliability of pain intensity and pain unpleasantness ratings related to pressure-pain thresholds (PPTs) of just noticeable pain (JNP), weak pain (WP) and moderate pain (MP) among healthy individuals.

Methods: Fourteen healthy participants (eight women, six men) participated in three sessions of testing at varied intervals over the course of 72?h. In session one, a multiple random staircase method using hydraulic pressure algometry was used to measure PPT of JNP, WP and MP on thumbnail bed. In session 2, ratings of pain intensity and pain unpleasantness were recorded when stimuli at levels corresponding to PPT of JNP, WP and MP were repeatedly applied before and after 20?min of no intervention.

Results: Interclass correlation coefficient (ICC) values for pain ratings of JNP, WP and MP in intra-session reliability were 0.810, 0.826 and 0.881, respectively, whereas the values were 0.817, 0.792 and 0.910, respectively, for inter-session reliability. ICC values for pain unpleasantness were also highly consistent and repeatable. Temporal summation of pain intensity and pain unpleasantness were not related to the repeated application of pressure stimuli.

Conclusions: The findings indicate that the pain intensity and pain unpleasantness ratings for stimuli at levels equal to the thresholds of JNP, WP and MP have good intra- and inter-session reliability.

Significance: This study showed that both pain intensity and pain unpleasantness of JNP, WP and MP have good intra- and inter-session reliability and agreement. Furthermore, the temporal summation of pain or unpleasantness is not related to repeated application of pressure stimuli.

Abbreviations: JNP: Just noticeable pain; WP: Weak pain; MP: Moderate pain; PPTs: pressure-pain thresholds; HPA: Hydraulic pressure algometry; MRSM: multiple random staircase method     

Test–retest stability of the oral niacin test and electrodermal activity in patients with schizophrenia

In schizophrenia, well-replicated findings support an attenuated niacin skin-flush response. We have previously reported a delayed skin-flush after niacin ingestion and also an association between niacin non-responding and electrodermal non-responding in schizophrenia. The stability of the niacin and electrodermal tests was now studied in a test–retest design. An additional aim was to assess the association previously found.Twenty-three patients with schizophrenia underwent two sessions 3 months apart during which an oral niacin test was conducted and electrodermal activity was measured. Despite similar values for niacin outcome variables at the group level, there was high intraindividual variation. Test–retest stability for the oral niacin test was thus low, although a trend toward correlation for the dichotomous response criterion was found. Most electrodermal measures correlated between baseline and retest. A significant association between the tests was again found; niacin non-responding implied electrodermal non-responding, providing further support for a common underlying aberration in schizophrenia.     

Test–retest reliability of quadriceps muscle function outcomes in patients with knee osteoarthritis

Quadriceps muscle weakness and the underlying neuromuscular deficits have been increasingly studied over the last few years in patients with knee osteoarthritis, but the applied methodologies have never been validated for this specific population. The aim of this study was to investigate test–retest reliability of several quadriceps muscle function outcomes in patients with knee osteoarthritis both before and after knee arthroplasty surgery. Ten preoperative and 20 postoperative patients participated in two identical testing sessions. A series of voluntary and/or electrically stimulated contractions of the involved quadriceps with concomitant torque and electromyographic recordings were used to characterize muscle strength, muscle activation and muscle contraction properties. Vastus lateralis morphology (thickness and fascicle pennation angle) was also assessed using ultrasonography. Overall, good reliability scores were observed for the majority of the 13 assessed variables (nine variables with intraclass correlation coefficients >0.75, 12 variables with coefficients of variation <15%). The most reliable testing protocol for patients with knee osteoarthritis would entail the assessment of (1) isometric maximal voluntary torque for evaluating muscle strength, with (2) simultaneous vastus lateralis electromyographic activity for evaluating muscle activation, (3) potentiated (resting) doublet peak torque for evaluating muscle contractility, and (4) vastus lateralis thickness for evaluating muscle size.     

An efficiency analysis of high-order combinations of gene–gene interactions using multifactor-dimensionality reduction

Background

Multifactor dimensionality reduction (MDR) is widely used to analyze interactions of genes to determine the complex relationship between diseases and polymorphisms in humans. However, the astronomical number of high-order combinations makes MDR a highly time-consuming process which can be difficult to implement for multiple tests to identify more complex interactions between genes. This study proposes a new framework, named fast MDR (FMDR), which is a greedy search strategy based on the joint effect property.

Results

Six models with different minor allele frequencies (MAFs) and different sample sizes were used to generate the six simulation data sets. A real data set was obtained from the mitochondrial D-loop of chronic dialysis patients. Comparison of results from the simulation data and real data sets showed that FMDR identified significant gene–gene interaction with less computational complexity than the MDR in high-order interaction analysis.

Conclusion

FMDR improves the MDR difficulties associated with the computational loading of high-order SNPs and can be used to evaluate the relative effects of each individual SNP on disease susceptibility. FMDR is freely available at http://bioinfo.kmu.edu.tw/FMDR.rar.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1717-8) contains supplementary material, which is available to authorized users.     

Thermal analysis of the dentine tubule under hot and cold stimuli using fluid–structure interaction simulation

The objective of this study is to compare the thermal stress changes in the tooth microstructures and the hydrodynamic changes of the dental fluid under hot and cold stimuli. The dimension of the microstructures of eleven cats’ teeth was measured by scanning electron microscopy, and the changes in thermal stress during cold and hot stimulation were calculated by 3D fluid–structure interaction modeling. Evaluation of results, following data validation, indicated that the maximum velocities in cold and hot stimuli were ??410.2?±?17.6 and +?205.1?±?8.7 µm/s, respectively. The corresponding data for maximum thermal stress were ??20.27?±?0.79 and +?10.13?±?0.24 cmHg, respectively. The thermal stress caused by cold stimulus could influence almost 2.9 times faster than that caused by hot stimulus, and the durability of the thermal stress caused by hot stimulus was 71% greater than that by cold stimulus under similar conditions. The maximum stress was on the tip of the odontoblast, while the stress in lateral walls of the odontoblast and terminal fibril was very weak. There is hence a higher possibility of pain transmission with activation of stress-sensitive ion channels at the tip of the odontoblast. The maximum thermal stress resulted from the cold stimulus is double that produced by the hot stimulus. There is a higher possibility of pain transmission in the lateral walls of the odontoblast and terminal fibril by releasing mediators during the cold stimulation than the hot stimulation. These two reasons can be associated with a greater pain sensation due to intake of cold liquids.     

A reliability measure of protein–protein interactions and a reliability measure-based search engine

Many methods developed for estimating the reliability of protein–protein interactions are based on the topology of protein–protein interaction networks. This paper describes a new reliability measure for protein–protein interactions, which does not rely on the topology of protein interaction networks, but expresses biological information on functional roles, sub-cellular localisations and protein classes as a scoring schema. The new measure is useful for filtering many spurious interactions, as well as for estimating the reliability of protein interaction data. In particular, the reliability measure can be used to search protein–protein interactions with the desired reliability in databases. The reliability-based search engine is available at http://yeast.hpid.org. We believe this is the first search engine for interacting proteins, which is made available to public. The search engine and the reliability measure of protein interactions should provide useful information for determining proteins to focus on.     

The reliability of Michaelis constants and maximum velocities estimated by using the integrated Michaelis–Menten equation

1. Experimental progress curves were simulated for a reaction obeying Michaelis-Menten kinetics. 2. K(m) and V were estimated (a) by fitting the integrated Michaelis-Menten equation to the progress curves, and (b) from the initial slopes of the curves (i.e. from initial velocities). 3. The integrated equation could not be fitted successfully by a non-linear method, so it was transformed and fitted by a linear method. 4. Provided that the initial substrate concentration was greater than K(m) and the data were precise enough, the integrated equation gave parameter estimates which were unbiased and as reliable as those derived from initial velocities although based on fewer experiments. 5. The integrated equation could be used for progress curves of unknown origin.     

CGRPα-expressing sensory neurons respond to stimuli that evoke sensations of pain and itch

Calcitonin gene-related peptide (CGRPα, encoded by Calca) is a classic marker of nociceptive dorsal root ganglia (DRG) neurons. Despite years of research, it is unclear what stimuli these neurons detect in vitro or in vivo. To facilitate functional studies of these neurons, we genetically targeted an axonal tracer (farnesylated enhanced green fluorescent protein; GFP) and a LoxP-stopped cell ablation construct (human diphtheria toxin receptor; DTR) to the Calca locus. In culture, 10-50% (depending on ligand) of all CGRPα-GFP-positive (+) neurons responded to capsaicin, mustard oil, menthol, acidic pH, ATP, and pruritogens (histamine and chloroquine), suggesting a role for peptidergic neurons in detecting noxious stimuli and itch. In contrast, few (2.2±1.3%) CGRPα-GFP(+) neurons responded to the TRPM8-selective cooling agent icilin. In adult mice, CGRPα-GFP(+) cell bodies were located in the DRG, spinal cord (motor neurons and dorsal horn neurons), brain and thyroid-reproducibly marking all cell types known to express Calca. Half of all CGRPα-GFP(+) DRG neurons expressed TRPV1, ～25% expressed neurofilament-200, <10% contained nonpeptidergic markers (IB4 and Prostatic acid phosphatase) and almost none (<1%) expressed TRPM8. CGRPα-GFP(+) neurons innervated the dorsal spinal cord and innervated cutaneous and visceral tissues. This included nerve endings in the epidermis and on guard hairs. Our study provides direct evidence that CGRPα(+) DRG neurons respond to agonists that evoke pain and itch and constitute a sensory circuit that is largely distinct from nonpeptidergic circuits and TRPM8(+)/cool temperature circuits. In future studies, it should be possible to conditionally ablate CGRPα-expressing neurons to evaluate sensory and non-sensory functions for these neurons.     

A comparison of enzyme-aided bleaching of softwood paper pulp using combinations of xylanase, mannanase and α-galactosidase

Enzymatic pretreatment of softwood kraft pulp was investigated using xylanase A (XylA) from Neocallimastix patriciarum in combination with mannanase and α-galactosidase. Mannanase A (ManA) from Pseudomonas fluorescens subsp. cellulosa and ManA from Clostridium thermocellum, both family 26 glycosyl hydrolases, are structurally diverse and exhibit different pH and temperature optima. Although neither mannanase was effective in pretreating softwood pulp alone, both enzymes were able to enhance the production of reducing sugar and the reduction of single-stage bleached κ number when used with the xylanase. Sequential incubations with XylA and P. fluorescens ManA produced the largest final κ number reduction in comparison to control pretreated pulp. The release of galactose from softwood pulp by α-galactosidase A (AgaA) from P. fluorescens was enhanced by the presence of ManA from the same microorganism, and a single pretreatment with these enzymes, in combination with XylA, gave the most effective κ number reduction using a single incubation. Results indicated that mixtures of hemicellulase activities can be chosen to enhance pulp bleachability. Received: 16 August 1999 / Received revision: 13 December 1999 / Accepted: 15 December 1999     

Chromatographic profiling of Pancharishta at different stages of its development using HPTLC,HPLC, GC–MS and UPLC–MS

Pancharishta is the traditional Ayurvedic polyherbal formulation prepared by decoction of plant materials followed by fermentation for preservation and facilitation of extraction due to the production of alcohol. Since the preparation of pancharishta involves various steps. The aim of the current investigation was to carry out comparative metabolomics profiling at different stages of preparation for the understanding impact of different steps and ingredients. A decoction of 21 plant materials are main components in pancharishta formulations followed by fermentation and addition of other ingredients with or without fermentation yielded eight different formulations. The vacuum concentration of pancharishta samples yielded a semisolid mass of different formulations ranging from 8 to 37% w/v. The HPTLC fingerprinting analysis of samples was carried out in butanol: ethanol: 0.5% v/v ammonia (5:4:0.5, v/v/v). Derivatization with anisaldehyde-sulphuric acid showed the presence of two major peaks at R_f 0.29 and 0.35. The peak at R_f 0.29 is intense in a formulation containing 12 extra plant materials. Quantification of gallic acid, ellagic acid, tannic acid, kaemferol and quercetin were carried out on newly developed HPLC method using acetonitrile and 0.5% v/v formic acid with a gradient elution. A significant difference in their content was found in different formulations. Further, polar and nonpolar metabolites of pancharishtha were analyzed using UPLC–MS and GC–MS, respectively. GC–MS profiling results in the identification of 144 metabolites among them 26 are common metabolites at different stages. The UPLC–MS analysis resulted in the tentative identification of 43 metabolites. The results of UPLC–MS and GC–MS analysis were used for multivariate analysis using XLSTAT. Principal Component Analysis plot distributed all samples into four different clusters with two formulations each.     

Evaluation of five resynchronization methods using different combinations of PGF2α, GnRH, estradiol and an intravaginal progesterone device for insemination in Holstein cows

This study was conducted to determine the use of repeated transvaginal ultrasound-guided cumulus oocyte complex (COC) aspiration on COC recovery rate, in vitro embryo production (IVP) and subsequent pregnancy rates in Holstein Friesian (HF) and Aberdeen Angus (AA) cows (Experiment 1), and in pregnant and non-pregnant Holstein Friesian cows (Experiment 2). Cycling, non-pregnant HF (n=17) and AA (n=32) cows with 40-70 days postpartum, between 3 and 5 years of age were used in the Experiment 1. All cows were submitted to repeated transvaginal ultrasound-guided COC aspiration twice a week for 5-7 weeks. Cumulus ooctye complexes (COC) were in vitro matured, fertilized and cultured for 8 days. An overall of 100 and 350 embryos from HF and AA cows respectively were cryopreserved using a conventional slow freezing (Experiment 1). A total of 81 and 285 frozen-thawed embryos from HF and AA cows respectively were transferred to recipient cows. Pregnancy diagnosis was performed at 60 and 150 days of gestation using transrectal ultrasonography. In Experiment 2, cycling non-pregnant (n=9) and pregnant (n=8) HF cows were submitted to repeated ultrasound-guided COC aspiration and COC were in vitro matured, fertilized and cultured as in Experiment 1, except that embryos were cryopreserved but not thawed and transferred as described for Experiment 1. The results of this study indicate that COC recovery rate and blastocyts production are affected by the breed of the donor cow. The quality of blastocyts produced from both breed did not differ in terms of pregnancy and calving rates (Experiment 1). The physiologic state of pregnancy did not affect COC recovery rate and blastocysts production per donor/session (Experiment 2). The use of ultrasound-guided COC aspiration and IVP could be a powerful technique to improve the genetic of beef and dairy cattle managed under pasture-based conditions management in the southern Chile.     

Identification of sterility-inducing cytoplasms in rye using the plasmotype–genotype interaction test and newly developed SCAR markers

A series of 25 rye (Secale cereale L.) inbred lines was tested with respect to three mitochondrial sequence-characterized amplified region (SCAR) polymorphisms. The analysis revealed a close association between the marker-determined mitotypes and plasmotypes (cytoplasm types known from breeding data) represented by the inbreds. The mitochondrial markers also confirmed normal (N-) cytoplasmic character of three wild rye species: Secale montanum, S. vavilovii and S. kuprijanovii. For 186 plants from open-pollinated cultivars of Turkish and South American origin, cytoplasm identification was performed with the use of crossing with double non-restoring tester. In 77 plants the normal (N) cytoplasm was detected, and for 63 of these the PCR analysis was performed producing results which were consistent with the genetic data based on testcrossing and phenotype assessment. The mitochondrial markers also confirmed a presence of sterility-inducing cytoplasm in the remaining 109 plants. Moreover, the markers allowed for differentiation between Pampa (P-) and Vavilovii (V-) cytoplasmic individuals. For 60 plants the latter results were verified using crosses with a line maintaining P-cytoplasmic sterility and acting as a restorer of the V-cytoplasm. For two of these plants contradicting results were produced with the applied methods of cytoplasm identification and the basis of this discrepancy is discussed. Regardless of the identification method, widespread occurrence of a sterility-inducing cytoplasm was revealed, especially in South American populations.