Lung hyperinflation and functional exercise capacity in patients with COPD – a three-year longitudinal study

Background

Lung hyperinflation contributes to dyspnea, morbidity and mortality in chronic obstructive pulmonary disease (COPD). The inspiratory-to-total lung capacity (IC/TLC) ratio is a measure of lung hyperinflation and is associated with exercise intolerance. However, knowledge of its effect on longitudinal change in the 6-min walk distance (6MWD) in patients with COPD is scarce. We aimed to study whether the IC/TLC ratio predicts longitudinal change in 6MWD in patients with COPD.

Methods

This prospective cohort study included 389 patients aged 40–75?years with clinically stable COPD in Global Initiative for Chronic Obstructive Lung Disease stages II-IV. The 6MWD was measured at baseline, and after one and 3 years. We performed generalized estimating equation regression analyses to examine predictors for longitudinal change in 6MWD. Predictors at baseline were: IC/TLC ratio, age, gender, pack years, fat mass index (FMI), fat-free mass index (FFMI), number of exacerbations within 12?months prior to inclusion, Charlson index for comorbidities, forced vital capacity (FVC), forced expiratory volume in 1?s (FEV₁), and light and hard self-reported physical activity.

Results

Reduced IC/TLC ratio (p?<?0.001) was a statistically significant predictor for decline in 6MWD. With a 0.1-unit decrease in baseline IC/TLC ratio, the annual decline in 6MWD was 12.7?m (p?<?0.001). Study participants with an IC/TLC ratio in the upper quartiles maintained their 6MWD from baseline to year 3, while it was significantly reduced for the patients with an IC/TLC ratio in the lower quartiles. Absence of light and hard physical activity, increased age and FMI, decreased FEV₁ and FVC, more frequent exacerbations and higher Charlson comorbidity index were also predictors for lower 6MWD at any given time, but did not predict higher rate of decline over the timespan of the study.

Conclusion

Our findings demonstrated that patients with less lung hyperinflation at baseline maintained their functional exercise capacity during the follow-up period, and that it was significantly reduced for patients with increased lung hyperinflation.

     

Lost in the world of functional genomics, systems biology, and translational research: is there life after the Milstein award?

We have always wanted to save the world from the scourges of virus infection by developing better drugs and vaccines. But fully understanding the intricacies of virus–host interactions, the first step in achieving this goal, requires the ability to view the process on a grand scale. The advent of high-throughput technologies, such as DNA microarrays and mass spectrometry, provided the first opportunities to obtain such a view. Here, we describe our efforts to use these tools to focus on the changes in cellular gene expression and protein abundance that occur in response to virus infection. By examining these changes in a comprehensive manner, we have been able to discover exciting new insights into innate immunity, interferon and cytokine signaling, and the strategies used by viruses to overcome these cellular defenses. Functional genomics may yet save the world from killer viruses.     

Is oral microbiome of children able to maintain resistance and functional stability in response to short-term interference of ingesta?

Dear Editor, A series of studies had focused on the ecological stability of human microbiome (Lozupone et al.,2012;Faith et al.,2013;Moya and Ferrer,2016).Despite the continuous perturbation and the highly personalized composition within the human microbiome (Human Microbiome Project,2012),healthy adults stably maintain their microbial communities in terms of space and time (Faith et al.,2013;Moya and Ferrer,2016;Oh et al.,2016).This stability is proved to be critical for the well-being of human body (Lozupone et al.,2012).On the contrary,major shifts in microbial community composition are often related to diseases (Lynch and Pedersen,2016).     

Do the phenology and functional stem attributes of woody species allow for the identification of functional groups in the semiarid region of Brazil?

The phenology of tree species in environments that are subject to strong climatic seasonality is mainly determined by water availability, which may vary as a function of wood density. The relationship among phenology, water potential, wood density and the capacity of water storage in the stem were determined for woody species of caatinga vegetation (dry forest) in the semiarid region of NE Brazil. Leaf flush and fall, flowering and fruiting events were recorded over a 31-month period, and the water potential was measured over a two-year period. These data were related to precipitation, water availability in the soil and photoperiod. Seven deciduous species exhibited low wood density (DLWD,?<0.5?g?cm^?3), high capacity of water storage in the stem (until 250?% of the dry weight) and high water potential during the year, as opposed to 15 deciduous species that showed high wood density (DHWD,?≥0.5?g?cm^?3). Leaf flush, flowering and the fruiting of DHWD species were related to precipitation, whereas these phenological events occurred at the end of the dry season and/or the beginning of the rainy season for DLWD species and were related to the photoperiod. The two evergreen species showed variations of water potential that were intermediate between those of DHWD and DLWD deciduous species, leaf flush during the dry season and flowering at the end of dry season. These results suggest the existence of three functional groups: evergreen species, DHWD deciduous species and DLWD deciduous species.     

Do earthworms affect dynamics of functional response and genetic structure of microbial community in a lab-scale composting system?

Two laboratory-scale systems were set up (i) composting (without earthworms) and (ii) vermicomposting (with earthworms) and were monitored for 60 days after pre-composting. The physico-chemical parameters (pH, C/N, organic matter, NH(4)(+)-N and ash content) showed similar evolution in both systems except a higher NH(4)(+)-N in the initial vermicomposts. However, principle component analysis (PCA) of enzymatic activities and community level physiological profiles revealed differences in the functional response of microbial communities in compost and vermicompost during maturation. Dehydrogenase activity and bacterial counts indicated a steady decrease in biological activity and population during composting, whereas vermicomposting exhibited higher activity on day 30 and a reduction in bacterial counts on day 10. PCA of denatured gradient gel electrophoresis (DGGE) profiles showed divergent dynamics of bacterial communities in two processes. These results indicated differences in the functional response and genetic structure of microbial community in composts and vermicomposts despite similar changes in their physico-chemical parameters.     

Androgen deficiency in male patients diagnosed with ANCA-associated vasculitis: a cause of fatigue and reduced health-related quality of life?

Introduction

Low testosterone levels in men are associated with fatigue, limited physical performance and reduced health-related quality of life (HRQOL); however, this relationship has never been assessed in patients with anti-neutrophil cytoplasmic antibodies (ANCA) -associated vasculitides (AAV). The aim of this study was to assess the prevalence of androgen deficiency and to investigate the role of testosterone in fatigue, limited physical condition and reduced HRQOL in men with AAV.

Methods

Male patients with AAV in remission were included in this study. Fatigue and HRQOL were assessed by the multi-dimensional fatigue inventory (MFI)-20 and RAND-36 questionnaires.

Results

Seventy male patients with a mean age of 59 years (SD 12) were included. Scores of almost all subscales of both questionnaires were significantly worse in patients compared to controls. Mean total testosterone and free testosterone levels were 13.8 nmol/L (SD 5.6) and 256 pmol/L (SD 102), respectively. Androgen deficiency (defined according to Endocrine Society Clinical Practice Guidelines) was present in 47% of patients. Scores in the subscales of general health perception, physical functioning and reduced activity were significantly worse in patients with androgen deficiency compared to patients with normal androgen levels. Testosterone and age were predictors for the RAND-36 physical component summary in multiple linear regression analysis. Testosterone, age, vasculitis damage index (VDI) and C-reactive protein (CRP) were associated with the MFI-20 subscale of general fatigue.

Conclusions

This study showed that androgen deficiency was present in a substantial number of patients with AAV. Testosterone was one of the predictors for physical functioning and fatigue. Testosterone may play a role in fatigue, reduced physical performance and HRQOL in male patients with AAV.     

Long-term effects of oxandrolone treatment in childhood on neurocognition,quality of life and social–emotional functioning in young adults with Turner syndrome

Turner syndrome (TS) is the result of (partial) absence of one X-chromosome. Besides short stature, gonadal dysgenesis and other physical aspects, TS women have typical psychological features. Since psychological effects of androgen exposure in childhood probably are long-lasting, we explored long-term psychological functioning after oxandrolone (Ox) therapy during childhood in adults with TS in terms of neurocognition, quality of life and social–emotional functioning. During the initial study, girls were treated with growth hormone (GH) combined with placebo (Pl), Ox 0.03 mg/kg/day, or Ox 0.06 mg/kg/day from the age of eight, and estrogen from the age of twelve. Sixty-eight women participated in the current double-blinded follow-up study (mean age 24.0 years, mean time since stopping GH/Ox 8.7 years). We found no effects on neurocognition. Concerning quality of life women treated with Ox had higher anxiety levels (STAI 37.4 ± 8.4 vs 31.8 ± 5.0, p = 0.002) and higher scores on the depression subscale of the SCL-90-R (25.7 ± 10.7 vs 20.5 ± 4.7, p = 0.01). Regarding social–emotional functioning, emotion perception for fearful faces was lower in the Ox-treated patients, without effect on interpersonal behavior. Our exploratory study is the first to suggest that androgen treatment in adolescence possibly has long-term effects on adult quality of life and social–emotional functioning. However, differences are small and clinical implications of our results seem limited. Therefore we would not recommend against the use of Ox in light of psychological consequences.     

Do broiler chicks have a cognitive representation of food quality?. Appetitive, behavioural and ingestive responses to a change in diet quality

It has been observed that when a new diet formulation, or a new batch of the same diet formulation, is presented to poultry, there is a transitory suppression of feeding. It appears that the birds do not recognise the food as being edible, or classify it as being 'unknown'. In order to understand more about food recognition and rejection, the aim of this experiment was to determine whether cognitive processes are involved. Sixteen groups of four broiler chicks were used, and were fed a low quality diet in their home pens. The groups of chicks were trained to run a winding maze to gain access to a high quality diet in a test trough for 15min per day. When training was completed, the feed in the test trough for eight of the groups was changed to that which they received in the home pen, while the other eight groups received no change as a control. Time to traverse the runway did not show an immediate decrease on the day after the change (P>0.05) as would be expected if the birds used a cognitive comparison to determine speed of approach to the test trough. However, the experimental groups were significantly slower compared to the control groups after 4 days (P<0.05). The behaviour observed on the day of the change was indicative of frustration with more scratching and hurried movements shown (P<0.05), and less pecking at feed (P<0.01). Food consumption was lower for experimental groups compared to control groups on all days from the day of change onward (P<0.001). It was concluded that although there was no definitive evidence for the presence of a cognitive representation of food, this may have been due in part to the testing of groups of birds rather than individuals, and the way in which food quality is perceived. The occurrence of behaviours indicative of frustration suggest that a cognitive expectation may have been present.     

Do short courses in evidence based medicine improve knowledge and skills? Validation of Berlin questionnaire and before and after study of courses in evidence based medicine

ObjectiveTo develop and validate an instrument for measuring knowledge and skills in evidence based medicine and to investigate whether short courses in evidence based medicine lead to a meaningful increase in knowledge and skills.DesignDevelopment and validation of an assessment instrument and before and after study.SettingVarious postgraduate short courses in evidence based medicine in Germany.ParticipantsThe instrument was validated with experts in evidence based medicine, postgraduate doctors, and medical students. The effect of courses was assessed by postgraduate doctors from medical and surgical backgrounds.InterventionIntensive 3 day courses in evidence based medicine delivered through tutor facilitated small groups.ResultsThe questionnaire distinguished reliably between groups with different expertise in evidence based medicine. Experts attained a threefold higher average score than students. Postgraduates who had not attended a course performed better than students but significantly worse than experts. Knowledge and skills in evidence based medicine increased after the course by 57% (mean score before course 6.3 (SD 2.9) v 9.9 (SD 2.8), P<0.001). No difference was found among experts or students in absence of an intervention.ConclusionsThe instrument reliably assessed knowledge and skills in evidence based medicine. An intensive 3 day course in evidence based medicine led to a significant increase in knowledge and skills.

What is already known on this topic

Numerous observational studies have investigated the impact of teaching evidence based medicine to healthcare professionals, with conflicting resultsMost of the studies were of poor methodological quality

What this study adds

An instrument assessing basic knowledge and skills required for practising evidence based medicine was developed and validatedAn intensive 3 day course on evidence based medicine for doctors from various backgrounds and training level led to a clinically meaningful improvement of knowledge and skills     

Do the capacity and kinetics for modification of xanthophyll cycle pool size depend on growth irradiance in temperate trees?

The present study investigated the interaction of growth irradiance (Q_int) with leaf capacity for and kinetics of adjustment of the pool size of xanthophyll cycle carotenoids (sum of violaxanthin, antheraxanthin and zeaxanthin; VAZ) and photosynthetic electron transport rate (J_max) after changes in leaf light environment. Individual leaves of lower‐canopy/lower photosynthetic capacity species Tilia cordata Mill. and upper canopy/higher photosynthetic capacity species Populus tremula L. were either illuminated by additional light of 500–800 µmol m^?2 s^?1 for 12 h photoperiod or enclosed in shade bags. The extra irradiance increased the total amount of light intercepted by two‐fold for the upper and 10–15‐fold for the lower canopy leaves, whereas the shade bags transmitted 45% of incident irradiance. In control leaves, VAZ/area, VAZ/Chl and J_max were positively associated with leaf growth irradiance (Q_int). After 11 d extra illumination, VAZ/Chl increased in all cases due to a strong reduction in foliar chlorophyll, but VAZ/area increased in the upper canopy leaves of both species, and remained constant or decreased in the lower canopy leaves of T. cordata. The slope for VAZ/area changes with cumulative extra irradiance was positively associated with Q_int only in T. cordata, but not in P. tremula. Nevertheless, all leaves of P. tremula increased VAZ/area more than the most responsive leaves of T. cordata. Shading reduced VAZ content only in P. tremula, but not in T. cordata, again demonstrating that P. tremula is a more responsive species. Compatible with the hypothesis of the role of VAZ in photoprotection, the rates of photosynthetic electron transport declined less in P. tremula than in T. cordata after the extra irradiance treatment. However, foliar chlorophyll contents of the exposed leaves declined significantly more in the upper canopy of P. tremula, which is not consistent with the suggestion that the leaves with the highest VAZ content are more resistant to photoinhibition. This study demonstrates that previous leaf light environment may significantly affect the adaptation capacity of foliage to altered light environment, and also that species differences in photosynthetic capacity and acclimation potentials importantly alter this interaction.     

Do functional demands associated with locomotor habitat,diet, and activity pattern drive skull shape evolution in musteloid carnivorans?

A major goal of evolutionary studies is to better understand how complex morphologies are related to the different functions and behaviours in which they are involved. For example, during locomotion and hunting behaviour, the head and the eyes have to stay at an appropriate level in order to reliably judge distance as well as to provide postural information. The morphology and orientation of the orbits and cranial base will have an impact on eye orientation. Consequently, variation in orbital and cranial base morphology is expected to be correlated with aspects of an animal's lifestyle. In this study, we investigate whether the shape of the skull evolves in response to the functional demands imposed by ecology and behaviour using geometric morphometric methods. We test if locomotor habitats, diet, and activity pattern influence the shape of the skull in musteloid carnivorans using (M)ANOVAs and phylogenetic (M)ANOVAs, and explore the functional correlates of morphological features in relation to locomotor habitats, diet, and activity pattern. Our results show that phylogeny, locomotion and, diet strongly influence the shape of the skull, whereas the activity pattern seems to have a weakest influence. We also show that the locomotor environment is highly integrated with foraging and feeding, which can lead to similar selective pressures and drive the evolution of skull shape in the same direction. Finally, we show similar responses to functional demands in musteloids, a super family of close related species, as are typically observed across all mammals suggesting the pervasiveness of these functional demands.     

A case of non-scaling in mammalian physiology? Body size, digestive capacity, food intake, and ingesta passage in mammalian herbivores

As gut capacity is assumed to scale linearly to body mass (BM), and dry matter intake (DMI) to metabolic body weight (BM(0.75)), it has been proposed that ingesta mean retention time (MRT) should scale to BM(0.25) in herbivorous mammals. We test these assumptions with the most comprehensive literature data collations (n=74 species for gut capacity, n=93 species for DMI and MRT) to date. For MRT, only data from studies was used during which DMI was also recorded. Gut capacity scaled to BM(1.06). In spite of large differences in feeding regimes, absolute DMI (kg/d) scaled to BM(0.76) across all species tested. Regardless of this allometry inherent in the dataset, there was only a very low allometric scaling of MRT with BM(0.14) across all species. If species were divided according to the morphophysiological design of their digestive tract, there was non-significant scaling of MRT with BM(0.04) in colon fermenters, BM(0.08) in non-ruminant foregut fermenters, BM(0.06) in browsing and BM(0.04) in grazing ruminants. In contrast, MRT significantly scaled to BM(0.24) (CI 0.16-0.33) in the caecum fermenters. The results suggest that below a certain body size, long MRTs cannot be achieved even though coprophagy is performed; this supports the assumption of a potential body size limitation for herbivory on the lower end of the body size range. However, above a 500 g-threshold, there is no indication of a substantial general increase of MRT with BM. We therefore consider ingesta retention in mammalian herbivores an example of a biological, time-dependent variable that can, on an interspecific level, be dissociated from a supposed obligatory allometric scaling by the morphophysiological design of the digestive tract. We propose that very large body size does not automatically imply a digestive advantage, because long MRTs do not seem to be a characteristic of very large species only. A comparison of the relative DMI (g/kg(0.75)) with MRT indicates that, on an interspecific level, higher intakes are correlated to shorter MRTs in caecum, colon and non-ruminant foregut fermenters; in contrast, no significant correlation between relative DMI and MRT is evident in ruminants.     

Consumption of flavonoid-rich foods and increased plasma antioxidant capacity in humans: cause, consequence, or epiphenomenon?

Increased fruit and vegetable consumption is associated with a decreased incidence of cardiovascular diseases, cancer, and other chronic diseases. The beneficial health effects of fruits and vegetables have been attributed, in part, to antioxidant flavonoids present in these foods. Large, transient increases in the total antioxidant capacity of plasma have often been observed after the consumption of flavonoid-rich foods by humans. These observations led to the hypothesis that dietary flavonoids play a significant role as antioxidants in vivo, thereby reducing chronic disease risk. This notion, however, has been challenged recently by studies on the bioavailability of flavonoids, which indicate that they reach only very low concentrations in human plasma after the consumption of flavonoid-rich foods. In addition, most flavonoids are extensively metabolized in vivo, which can affect their antioxidant capacity. Furthermore, fruits and vegetables contain many macro- and micronutrients, in addition to flavonoids, that may directly or through their metabolism affect the total antioxidant capacity of plasma. In this article, we critically review the published research in this field with the goal to assess the contribution of dietary flavonoids to the total antioxidant capacity of plasma in humans. We conclude that the large increase in plasma total antioxidant capacity observed after the consumption of flavonoid-rich foods is not caused by the flavonoids themselves, but is likely the consequence of increased uric acid levels.     

Do sodium and potassium forms of Na,K-ATPase differ in their secondary structure?

Infrared spectroscopy in the amide I region of purified membrane-bound Na,K-ATPase preparation shows that Na+- and K+-bound forms of the enzyme have almost the same secondary structure. No difference is detected in the beta-structure (pleated sheets) content. This is contrary to the statement of the recent paper (Gresalfi, T. J., and Wallace, B. A. (1984) J. Biol. Chem. 259, 2622-2628) where a similar preparation was examined by circular dichroism spectroscopy and it was claimed that net 7% of protein peptide groups undergo a beta-sheet to alpha-helix conformational change upon Na,K-ATPase conversion from the K+ to the Na+ form. The discrepancy of the results is most likely caused by the particulate nature of the enzyme preparations used that could lead to optical artifacts in CD but not in IR measurements. A thorough comparison of IR spectra of these enzyme forms has revealed a very minor spectral difference which could suggest conformational perturbations, if any, of a much lower scale and another type than that claimed by Gresalfi and Wallace. The K+ form tends to absorb slightly more in the region of the alpha-helix band. This could reflect some distortion or a transition to a random coil structure of a small fraction of alpha-helical segments (less than or equal to 2% protein peptide groups) upon the enzyme conversion from the K+ to the Na+ form.     

Where Do We Go from Here? Prevalence of Trachoma Three Years after Stopping Mass Distribution of Antibiotics in the Regions of Kayes and Koulikoro,Mali

Objectives

A national survey in 1997 demonstrated that trachoma was endemic in Mali. Interventions to control trachoma including mass drug administration (MDA) with azithromycin were launched in the regions of Kayes and Koulikoro in 2003. MDA was discontinued after three annual rounds in 2006, and an impact survey conducted. We resurveyed all districts in Kayes and Koulikoro in 2009 to reassess trachoma prevalence and determine intervention objectives for the future. In this paper we present findings from both the 2006 and 2009 surveys.

Methods

Population-based cluster surveys were conducted in each of the nine districts in Koulikoro in 2006 and 2009, whilst in Kayes, four of seven districts in 2006 and all seven districts in 2009 were surveyed. Household members present were examined for clinical signs of trachoma.

Results

Overall, 29,179 persons from 2,528 compounds, in 260 clusters were examined in 2006 and 32,918 from 7,533 households in 320 clusters in 2009. The prevalence of TF in children aged 1–9 years in Kayes and Koulikoro was 3.9% (95%CI 2.9–5.0%, range by district 1.2–5.4%) and 2.7% (95%CI 2.3–3.1%, range by district 0.1–5.0%) respectively in 2006. In 2009 TF prevalence was 7.26% (95%CI 6.2–8.2%, range by district 2.5–15.4%) in Kayes and 8.19% (95%CI 7.3–9.1%, range by district 1.7–17.2%) in Koulikoro among children of the same age group. TT in adults 15 years of age and older was 2.37% (95%CI 1.66–3.07%, range by district 0.30–3.54%) in 2006 and 1.37% (95%CI 1.02–1.72%, range by district 0.37–1.87%) in 2009 in Kayes and 1.75% (95%CI 1.31–2.23%, range by district 1.06–2.49%) in 2006 and 1.08% (95%CI 0.86–1.30%, range by district 0.34–1.78%) in 2009 in Koulikoro.

Conclusions

Using WHO guidelines for decision making, four districts, Bafoulabe in Kayes Region; and Banamba, Kolokani and Koulikoro in Koulikoro Region, still meet criteria for district-wide implementation of the full SAFE strategy as TF in children exceeds 10%. A community-by-community approach to trachoma control may now be required in the other twelve districts. Trichiasis surgery provision remains a need in all districts and should be enhanced in six districts in Kayes and five in Koulikoro where the prevalence exceeded 1.0% in adults. Since 1997 great progress has been observed in the fight against blinding trachoma; however, greater effort is required to meet the elimination target of 2015.     

Azathioprine, a clastogen in human somatic cells? Analysis of chromosome damage and SCE in lymphocytes after exposure in vivo and in vitro

Chromosomal analyses in lymphocytes of 28 patients with multiple sclerosis were carried out before, during and after Azathioprine (Aza) therapy. Only a higher incidence of gaps was found in treated patients than in a group of healthy persons but not in comparison with untreated patients. Similarly, no significant clastogenic effect was observed in vitro after short-term and long-term treatment of unstimulated and stimulated lymphocytes with concentrations of 1--100 microgram Aza per ml. Treatment of cultures with 0.0001--4.0 microgram/ml did not yield increased SCE frequencies. The absence of any significant clastogenic effect of therapeutic doses of Aza on human somatic cells is deduced from an evaluation of previously published data and from the present results.     

Do anthocyanins function as antioxidants in leaves? Imaging of H2O2 in red and green leaves after mechanical injury

Purified anthocyanin extracts show strong antioxidant properties in vitro, but it is not known whether they can scavenge reactive oxygen in living cells. The oxidative responses in red and green portions of Pseudowintera colorata leaf laminae were compared by the real‐time imaging of H₂O₂ in cells after mechanical injury. An oxidative burst was elicited almost immediately from chloroplasts in the palisade mesophyll, as evidenced using the fluorochromes dichlorofluorescein and scopoletin. H₂O₂ accumulated in green lamina regions for 10 min, and then decreased slowly. By contrast, red regions recovered rapidly, and maintained consistently low levels of H₂O₂. Infusion of cells with N‐acetyl‐l ‐cysteine accelerated the depletion of H₂O₂ from green regions. Wounded leaves ultimately developed a localized necrotic lesion and an intense anthocyanic band. The red regions were enriched in anthocyanins, flavonols, dihydroflavonols, and hydroxycinnamic acids. Only the anthocyanins were suitably located to account for the enhanced rates of H₂O₂ scavenging. The data support the hypothesis that red cells have elevated antioxidant capabilities in vivo.     

Azathioprine,a clastogen in human somatic cells? Analysis of chromosome damage and SCE in lymphocytes after exposure in vivo and in vitro

Chromosomal analyses in lymphocytes of 28 patients with multiple sclerosis were carried out before, during and after Azathioprine (Aza) therapy. Only a higher incidence of gaps was found in treated patients than in a group of healthy persons but not in comparison with untreated patients. Similarly, no significant clastogenic effect was observed in vitro after short-term and long-term treatment of unstimulated and stimulated lymphocytes with concentrations of 1–100 μg Aza per ml. Treatment of cultures with 0.0001–4.0 μg/ml did not yield increased SCE frequencies. The absence of any significant clastogenic effect of therapeutic doses of Aza on human somatic cells is deduced from an evaluation of previously published data and from the present results.