Subtle Negative Effects Research Articles

Fine scale genetics reveals the subtle negative effects of roads on an endangered bat

The effective management of species with small and fragmented populations requires an in-depth understanding of how the effects of human-induced habitat disturbance shape the structure and gene flow at fine spatial scales. Identification of putative environmental barriers that affect individual exchange among subpopulations is imperative to prevent extinction risks. Here, we investigated how landscape affects the gene flow and relatedness structure of a population of the endangered lesser horseshoe bat (Rhinolophus hipposideros). We also assessed the effects of sexbiased dispersal on genetic relatedness. We genotyped 287 bat samples collected across southern Portugal and developed resistance surfaces for landscape variables hypothesized to affect gene flow. Then, we used spatially explicit models to fit relatedness distance through the resistance surfaces. We found genetic evidence of sex-biased dispersal and identified a significant fine scale structuring in the relatedness regarding females, the philopatric sex. Males displayed uniform levels of relatedness throughout the landscape. The results indicated less relatedness between the female´ from roosts located on proximity of roads than in roosts away from roads. Also, when analysing the sexes together the relatedness on roosts separated by highway were subtly less related in comparison to those occurring on the same side. Roads seem to be major shapers of the contemporary population structure of females, regardless of being relatively recent structures in the landscape. Furthermore, the relatedness patterns detected suggested that high tree density among roosts and continuity of forest patches in broader surrounding areas, promotes the relatedness among individuals. Landscape heterogeneity among roosts slightly decreases genetic relatedness. Nevertheless, those relationships are still weak, suggesting that population structuring driven by those factors is slowly ongoing. Thus, effective management measures should focus on issues for promoting safe road passages and suitable habitat corridors, allowing for the exchange of individuals and gene flow among lesser horseshoe bat roosts.

The response of Nassella trichotoma (serrated tussock) seeds and seedlings to different levels of fire intensity

Context Fire is an important disturbance regime in grassland communities, since it is responsible for stimulating the regeneration of many species and for maintaining levels of biodiversity. When invasive plants, such as Nassella trichotoma, establish and become widespread in a grassland community, these important fire events can be altered in intensity and frequency, which means that they are able to facilitate the establishment of the exotic species. Therefore, before fire can be recommended as a suitable control technique for invasive species, or alternatively to be integrated into grassland restoration programs, understanding the response of the seeds of exotic species to high temperatures, such as those experienced during a fire, should be well understood. Aims Our aim was to identify their response to a gradient of temperatures associated with different levels of fire intensity. We examined how increased duration of exposure affects their response, and whether seed age or seed moisture content affect the germination response of this species. Methods To gain a fuller understanding of the fire response of N. trichotoma’s seedbank, seeds were collected in 2016, 2017, 2018 and 2019 and then stored until the commencement of the experiments in 2020. Selected seeds were first subjected to an increasing temperature gradient (80°C, 100°C, 120°C, 140°C, and a control), and an increasing duration of exposure (of 1, 3, 6, and 9 min). In the second experiment, one population was selected to test these same temperatures and duration of exposure after the seeds were hydrated to 15%, 50%, or 95%. Last, seedlings were grown for 3 months under glasshouse conditions and then exposed to increasing temperatures (20°C, 60°C, 80°C, 100°C, and 120°C), and an increasing duration of exposure (3, 6, and 9 min). The seedlings were assessed 2 weeks after the heat exposure for signs of damage. Key results It was found that increased temperatures and duration of exposure had a subtle negative effect on germination parameters, including reduced total germination and increased time to 50% germination. The 140°C treatment was seen to be a significant threshold because it killed all the seeds at any duration of exposure. A significant difference among the ages of each seed lot was observed to be a factor on the tested germination metrics, with the oldest tested population (2016) demonstrating the highest germination percentage, uniformity, and rate. Seed germination percentage was significantly reduced for seeds hydrated to 95% compared with the control treatment, whereas no significant difference was observed for the seeds hydrated to 15% and 50%. For the heat treatment of the seedlings, damage to the leaves was observed in the 80°C, 100°C, and 120°C treatments, with some plants in the 120°C treatment experiencing extensive damage prior to resprouting. No seedlings were killed at the tested temperatures. Conclusions Results of this study indicated that fire may be a useful tool for reducing seedbank density by killing a high proportion of the seeds on the soil surface, or located within the top 1 cm of the soil profile, but not for seeds buried more deeply. Efficacy of fire on surface and shallow-buried seeds is improved with high seed moisture content; however, these seeds buried below this depth are still protected by the soil from the lethal effects of temperature. Implications Fire implemented before seed set could be used to effectively kill a large proportion of N. trichotoma seeds. However, for more comprehensive control, it is recommended that chemical treatment is integrated with the fire treatment to improve the overall control efficiency.

Restoration of KMT2C/MLL3 in human colorectal cancer cells reinforces genome-wide H3K4me1 profiles and influences cell growth and gene expression

BackgroundThe histone 3 lysine 4 (H3K4) monomethylase KMT2C is mutated across several cancer types; however, the effects of mutations on epigenome organization, gene expression, and cell growth are not clear. A frequently recurring mutation in colorectal cancer (CRC) with microsatellite instability is a single nucleotide deletion within the exon 38 poly-A(9) repeat (c.8390delA) which results in frameshift preceding the functional carboxy-terminal SET domain. To study effects of KMT2C expression in CRC cells, we restored one allele to wild type KMT2C in the two CRC cell lines RKO and HCT116, which both are homozygous c.8390delA mutant.ResultsGene editing resulted in increased KMT2C expression, increased H3K4me1 levels, altered gene expression profiles, and subtle negative effects on cell growth, where higher dependence and stronger effects of KMT2C expression were observed in RKO compared to HCT116 cells. Surprisingly, we found that the two RKO and HCT116 CRC cell lines have distinct baseline H3K4me1 epigenomic profiles. In RKO cells, a flatter genome-wide H3K4me1 profile was associated with more increased H3K4me1 deposition at enhancers, reduced cell growth, and more differential gene expression relative to HCT116 cells when KMT2C was restored. Profiling of H3K4me1 did not indicate a highly specific regulation of gene expression as KMT2C-induced H3K4me1 deposition was found globally and not at a specific enhancer sub-set in the engineered cells. Although we observed variation in differentially regulated gene sets between cell lines and individual clones, differentially expressed genes in both cell lines included genes linked to known cancer signaling pathways, estrogen response, hypoxia response, and aspects of immune system regulation.ConclusionsHere, KMT2C restoration reduced CRC cell growth and reinforced genome-wide H3K4me1 deposition at enhancers; however, the effects varied depending upon the H3K4me1 status of KMT2C deficient cells. Results indicate that KMT2C inactivation may promote colorectal cancer development through transcriptional dysregulation in several pathways with known cancer relevance.

Changes in hypopharyngeal glands of nurse bees (Apis mellifera) induced by pollen-containing sublethal doses of the herbicide Roundup®

Decreasing pollinator populations worldwide has generated great concern and stimulated countless studies to understand the origin of colony losses. One main cause is the indiscriminate use of different pesticides, producing subtle negative effects on bee physiology and behavior. Royal jelly synthesized in the hypopharyngeal glands is an essential protein for feeding all individuals of the hive, especially the queen. Therefore, the present study aimed to determine the effect of sublethal concentrations of Roundup® on the hypopharyngeal glands of nursing workers, including its interference with the production of royal jelly. The herbicide was found to promote changes in the cellular ultrastructure of these glands, causing early degeneration of the rough endoplasmic reticulum and morphological and structural changes in the mitochondria. No changes were noted in the amount of royal jelly produced, but additional long-term studies are necessary to determine possible qualitative changes. This is the first study to evaluate the effect of Roundup® on the royal jelly-producing glands, showing that resultant alterations in these structures can trigger damage to the development and survival of bee colonies.

Effects of increasing pCO2 on life history traits and feeding of the littoral mysid Praunus flexuosus

Mysids, an important food web component in the littoral zone of coastal waters, have been neglected so far in ocean acidification research. Juveniles of the littoral mysid Praunus flexuosus were exposed in the laboratory to four pCO2 levels (530, 930, 1200, and 1600 µatm) for 5 weeks. In addition, juveniles were provided with two different food levels during the experiment. High pCO2 did not affect survival, but delayed moulting. Juvenile growth decreased and inter-moult period between the last moulting events increased with increasing pCO2 at low but not at high food supply, suggesting that high food availability is needed to prevent these negative effects of elevated pCO2. However, small individual juveniles showed lower feeding rates at high pCO2 compared to the control after prolonged exposure, suggesting decreased activity likely due to impaired metabolism. The subtle negative effects of elevated pCO2 on life history traits observed in this study suggest that P. flexuosus probably has to adapt to counteract adverse effects of predicted high pCO2, especially when food is limiting.

Subtle effects of the water soluble fraction of oil spills on natural phytoplankton assemblages enclosed in mesocosms

Four mesocosm experiments were conducted to evaluate the effect of episodic oil spills on coastal marine phytoplankton assemblages. The experimental design was selected to simulate the Prestige oil spill, which occurred in Galician coastal waters (NW Iberia) in November 2002. The empirical results indicate that no significant direct effects of the water soluble fraction of oil (20–60 μg l−1 of chrysene equivalents) on phytoplankton biomass and production were observed immediately after oil additions. Despite this, subtle negative effects on primary production were detected using a modelling approach, being the impact lower on phytoplankton communities dominated by diatoms. Consistent with the reduced direct effect of oil additions on phytoplankton biomass and photosynthesis-related variables, no indirect trophic cascading effects, previously reported in microcosm experiments, were detected. This shows that the effect of punctual inputs of the water accommodated fraction of oil on natural phytoplankton communities was very subtle, undetectable on some occasions, and of much lower magnitude than the effects recorded in microcosm experiments. This suggests that the initial composition of the phytoplankton community determines the degree of response and that the experimental approach adopted could explain the different, and sometimes contradictory, reported responses of the planktonic community to the input of oil into the marine environment.

Timing of prostaglandin F2α treatment in an estrogen-based protocol for timed artificial insemination or timed embryo transfer in lactating dairy cows

Objectives were to investigate progesterone concentrations and fertility comparing 2 different intervals from PGF2α treatment and induced ovulation in an estrogen-based ovulation synchronization protocol for timed artificial insemination (TAI) or timed embryo transfer (TET) in lactating dairy cows. A total of 1,058 lactating Holstein cows [primiparous (n=371) and multiparous (n=687)], yielding 34.1±0.33 kg of milk/d at various days in milk were randomly assigned to receive treatment with PGF2α on either d 7 or 8 of the following protocol: d 0: 2mg of estradiol benzoate + controlled internal drug release device; d 8: controlled internal drug release device removal + 1.0mg of estradiol cypionate; d 10: TAI or d 17: TET. Only cows with a corpus luteum at d 17 received an embryo and all cows received GnRH at TET. Pregnancy diagnoses were performed by detection (transrectal ultrasonography) of an embryo on d 28 or a fetus on d 60. Fertility [pregnancy per artificial insemination (P/AI) or pregnancy per embryo transfer (P/ET)] was affected by breeding technique (AI vs. ET) and time of PGF2α treatment (d 7 vs. 8) at the 28-d pregnancy diagnosis for TAI [32.9% (238) vs. 20.6% (168)] and TET cows [47% (243) vs. 40.7% (244)] and at the 60-d pregnancy diagnosis for TAI [30% (238) vs. 19.2% (168)] and TET cows [37.9% (243) vs. 33.5% (244)]. The progesterone (P4) concentration at d 10 altered fertility in TAI cows, with higher P/AI in cows with P4 concentration <0.1 ng/mL compared with cows with P4 concentration ≥0.1 ng/mL, and in ET cows, with higher P/ET in cows with P4 concentration <0.22 ng/mL compared with cows with P4 concentration ≥0.22 ng/mL. Prostaglandin F2α treatment at d 7 increased the percentage of cows with P4 <0.1 ng/mL on d 10 [39.4 (85) vs. 23.2 (54)]. Reducing the period between PGF2α and TAI from 72 to 48h in dairy cows resulted in a clear reduction in fertility in cows bred by TAI and a subtle negative effect in cows that received TET. The earlier PGF2α treatment benefits are most likely mediated through gamete transport, fertilization, or early embryo development and a more subtle effect of earlier PGF2α treatment that may be mediated through changes in the uterine or hormonal environment that manifests itself after ET on d 7.

Transcriptome profiling of Zymomonas mobilis under furfural stress

Furfural from lignocellulosic hydrolysates is the prevalent inhibitor to microorganisms during cellulosic ethanol production, but the molecular mechanisms of tolerance to this inhibitor in Zymomonas mobilis are still unclear. In this study, genome-wide transcriptional responses to furfural were investigated in Z. mobilis using microarray analysis. We found that 433 genes were differentially expressed in response to furfural. Furfural up- or down-regulated genes related to cell wall/membrane biogenesis, metabolism, and transcription. However, furfural has a subtle negative effect on Entner-Doudoroff pathway mRNAs. Our results revealed that furfural had effects on multiple aspects of cellular metabolism at the transcriptional level and that membrane might play important roles in response to furfural. This research has provided insights into the molecular response to furfural in Z. mobilis, and it will be helpful to construct more furfural-resistant strains for cellulosic ethanol production.

Soybean Flour and Wheat Germ Proportions in Artificial Diet and Their Effect on the Growth Rates of the Tobacco Budworm,Heliothis virescens

Soybean flour and wheat germ are the two most important protein components of wheat germ-based insect artificial diets. The effect of modifying the proportion of these two ingredients in a Noctuidae-specific diet was investigated utilizing the tobacco budworm Heliothis virescens (F.) (Lepidoptera: Noctuidae), with the goal of developing a suboptimal diet that, without drastically affecting this insect's growth and reproductive rates, could manifest subtle negative effects in this insect. The original diet formula contained 2.51% protein. When the proportions of soybean flour and wheat germ were changed to 2.15% protein the net reproductive rate of the first generation was significantly lower. In the second generation, the net reproductive rate, development time, percent female survivorship, fertility, intrinsic rate of increase, finite rate of increase and female longevity were significantly lower in both the 2.15% and 2.26% protein diets. The survival rate of immatures to the adult stage was 1% in the 2.05% protein diet in the first generation. Interestingly, females exposed to these suboptimal diets produced a significantly higher number of eggs but the survival of their larvae was significantly reduced. It is evident from these results that modifications to the protein content and the nutrient composition profile of the original wheat germ-based insect artificial formula can be used to produce subtle negative effects on the growth of tobacco budworm.

Clinical profile of sheep fed non-conventional feeds containing phenols and condensed tannins

A study was carried out to investigate the effects of feeding low quality non-conventional feeds (NCF) containing phenols and condensed tannins on the clinical profiles of sheep. Thirty-two Omani sheep were fed one of four diets with two base roughages, urea-treated palm frond (UTPF) and Rhodesgrass hay (RGH) and two concentrates, commercial concentrate (CC) and a by-products concentrate (BC) for 120 days. Haematological, serum biochemical and urine analyses were used to assess sheep health. Non-conventional feeds (urea-treated palm frond and by-products concentrate) contained higher levels of polyphenols and condensed tannins than conventional feeds (Rhodesgrass hay and commercial cubes). Feeds based on urea-treated palm frond had lower dry matter, crude protein, acid detergent fibre, neutral detergent fibre, gross energy ( P < 0.001) and ash ( P < 0.05) digestibility coefficients than those based on Rhodesgrass hay. Animals fed NCF had lower feed intake ( P < 0.001) and lower body gain ( P < 0.001) than those fed conventional ones. They also produced larger volumes of faeces ( P < 0.01) which contained higher levels of nitrogen ( P < 0.001) and had lower viscosity values of intestinal content ( P < 0.001). Rumen liquor of NCF-fed animals had higher pH and lower ammonia–nitrogen levels ( P < 0.01). Animals fed urea-treated palm frond plus by-products concentrate had lower lymphocyte ( P < 0.01), monocyte ( P < 0.05) and eosinophil ( P < 0.05) counts by the end of the trial than those fed Rhodesgrass hay based diets. The urea-treated palm frond and by-products concentrate fed animals had lower blood urea nitrogen (BUN) levels ( P < 0.05), higher ( P < 0.01) alanine aminotransferase (ALT) and lower serum iron ( P < 0.001) than those fed Rhodesgrass hay based diets. There was a trend of increasing blood, leukocytes and specific gravity in the urine of NCF-fed animals. This experiment implies that feeding low quality non-conventional feeds containing antinutritional factors for relatively long periods might produce subtle negative effects on the physiology and chemistry of the digestive system and blood parameters which might negatively affect sheep health and make them more susceptible to diseases.

PCBs and cognitive functioning of Mohawk adolescents

This paper reports on the relationships between the cognitive functioning and PCB current body burdens of adolescents in the Mohawk Nation of Akwesasne where there is concern about industrial pollution of the environment. Three cognitive tests (Woodcock Johnson-Revised, Test of Memory and Learning, and Ravens Progressive Matrices) provide 13 subtests that allow a variety of cognitive outcomes to be assessed. A summary measure of PCB level was created from the congeners detected in at least 50% of the participants. The most notable finding was the significant negative relationship between PCB levels and two separate measures of long term memory. There was also a negative relationship with a measure of comprehension and knowledge. Significant relationships were not large, but provide evidence of subtle negative effects of PCB exposure. This research was supported by NIEHS grants ES04913-10 and ES10904-05.

Characterization of the detachable Rho-dependent transcription terminator of the fimE gene in Escherichia coli K-12.

The fim genetic switch in the chromosome of Escherichia coli K-12 is an invertible DNA element that harbors the promoter for transcription of the downstream fim structural genes and a transcription terminator that acts on the upstream fimE regulatory gene. Switches oriented appropriately for structural gene transcription also allow fimE mRNA to read through, whereas those in the opposite orientation terminate the fimE message. We show here that termination is Rho dependent and is suppressed in a rho mutant or by bicyclomycin treatment when fimE mRNA is expressed by the fimE gene, either from a multicopy recombinant plasmid or in its native chromosomal location. Two cis-acting elements within the central portion of the 314-bp invertible DNA switch were identified as contributors to Rho-dependent termination and dissected. These fim sequence elements show similarities to well-characterized Rho utilization (rut) sites and consist of a boxA motif and a C-rich and G-poor region of approximately 40 bp. Deletion of the boxA motif alone had only a subtle negative effect on Rho function. However, when this element was deleted in combination with the C-rich, G-poor region, Rho function was considerably decreased. Altering the C-to-G ratio in favor of G in this portion of the switch also strongly attenuated transcription termination. The implications of the existence of a fimE-specific Rho-dependent terminator within the invertible switch are discussed in the context of the fim regulatory circuit.

Subthalamic nucleus stimulation affects striato-anterior cingulate cortex circuit in a response conflict task: a PET study.

The subthalamic nucleus (STN) has generally been considered as a relay station within frontal-subcortical motor control circuitry. Little is known about the influence of the STN on cognitive networks. Clinical observations and studies in animals suggest that the STN participates in non-motor functions which can now be probed in Parkinson's disease patients with deep brain stimulation of the STN, allowing selective and reversible modulation of this nucleus. Using PET, we studied changes in regional cerebral blood flow (rCBF) associated with a response conflict task (Stroop task) in Parkinson's disease patients ON and OFF bilateral STN stimulation. The Stroop task requires subjects to name the font colour of colour words (e.g. "blue") printed in an incongruent colour ink (e.g. yellow). During STN stimulation, impaired task performance (prolonged reaction times) was associated with decreased activation in both right anterior cingulate cortex (ACC) and right ventral striatum. Concomitant increased activation in left angular gyrus indicative of ongoing word processing during stimulation is consistent with an impairment to inhibit habitual responses. ACC and ventral striatum are part of the ACC circuit associated with response conflict tasks. The decreased activation during STN stimulation in the ACC circuit, while response conflict processing worsened, provides direct evidence of STN modulating non-motor basal ganglia-thalamocortical circuitry. Impairment in ACC circuit function could account for the subtle negative effects on cognition induced by STN stimulation.

Environmental exposure to polychlorinated biphenyls (PCBs) and dioxins

Polychlorinated biphenyl's (PCBs) and dioxins are environmental pollutants. Calculated on a body weight basis, prenatally as well as postnatally through breast‐feeding, large amounts are transferred from mother to the child. Formula is free of these substances. Considering their potential developmental neurotoxicity, we investigated long‐term effects of perinatal exposure to PCBs and dioxins on neurological and cognitive development. Methods. A group of 418 infants were followed from birth up to 6 years of age. Half of them were fully breast fed (BF) for at least 6 weeks. Prenatal PCB exposure was measured from cord and maternal blood. Postnatal exposure was reflected by PCB and dioxin levels in breast and formula milk and plasma PCB concentrations at 42 months of age. Both neurological and cognitive development were taken as outcome variables at 18, 42 months and at 6 years of age. At 18 and 42 months neurological condition was evaluated according to Hempel and Touwen and at 6 years of age according to Touwen. Condition was evaluated in terms of optimality. Separately, the fluency of movements was scored. Cognitive abilities were measured at 18 months by means of the Bayley Scales of Infant Development, at 42 months of age by the Kaufman Assessment Battery for Children (K‐ABC) and at 6 years of age by the McCarthy Scales. Results. At 18 months of age cognitive development was not affected by either pre‐ or postnatal exposure to the measured PCBs and dioxins. However, neurological examination showed an adverse effect of prenatal exposure to the measured pollutants on the neurological optimality score. At 42 months of age we found negative associations between prenatal PCB exposure and cognitive development. However, no effect was demonstrated of postnatal exposure to the measured pollutants. Neurological development was not affected by either pre‐ or postnatal exposure to PCBs and dioxins. At 6 years of age the preliminary results showed no evidence that cognitive and neurological development are affected by pre‐ and postnatal exposure to these pollutants. Despite a higher PCB exposure from breast milk we found at 18 months, 42 months of age, and at 6 years of age a beneficial effect of breast feeding on the quality of movements, in terms of fluency. Conclusion. These data support the evidence obtained from similar studies in the USA that prenatal exposure to PCBs may have subtle negative effects on early neurological and cognitive development of the child. Further studies are required to evaluate these findings in other parts of the world. Our studies showed consistently a beneficial effect of breast‐feeding on brain development from 18 months up to school age.

Endosulfan Exposure Disrupts Pheromonal Systems in the Red-Spotted Newt: A Mechanism for Subtle Effects of Environmental Chemicals

Because chemicals introduced into the environment by humans can affect both long-term survivorship and reproduction of amphibians, discovering the specific mechanisms through which these chemicals act may facilitate the development of plans for amphibian conservation. We investigated the amphibian pheromonal system as a potential target of common environmental chemicals. By treating female red-spotted newts, Notophthalmus viridescens, to a commonly used insecticide, endosulfan, we found that the pheromonal system is highly susceptible to low-concentration exposure. The impairment of the pheromonal system directly led to disrupted mate choice and lowered mating success. There were no other notable physiologic or behavioral changes demonstrated by the animals at the insecticide concentrations administered. Our findings suggest that the amphibian pheromonal system is one of the systems subject to subtle negative effects of environmental chemicals.

Endosulfan exposure disrupts pheromonal systems in the red-spotted newt: a mechanism for subtle effects of environmental chemicals.

Because chemicals introduced into the environment by humans can affect both long-term survivorship and reproduction of amphibians, discovering the specific mechanisms through which these chemicals act may facilitate the development of plans for amphibian conservation. We investigated the amphibian pheromonal system as a potential target of common environmental chemicals. By treating female red-spotted newts, Notophthalmus viridescens, to a commonly used insecticide, endosulfan, we found that the pheromonal system is highly susceptible to low-concentration exposure. The impairment of the pheromonal system directly led to disrupted mate choice and lowered mating success. There were no other notable physiologic or behavioral changes demonstrated by the animals at the insecticide concentrations administered. Our findings suggest that the amphibian pheromonal system is one of the systems subject to subtle negative effects of environmental chemicals.

Effect of passive smoking in pregnancy on neonatal nucleated red blood cells.

We evaluated whether the absolute nucleated red blood cell (RBC) count is elevated in term, appropriate for gestational age (AGA) infants born to women exposed to passive smoking in pregnancy. We compared absolute nucleated RBC counts taken during the first 12 hours of life in 2 groups of term, vaginally delivered infants, 1 group born to mothers who were routinely exposed to tobacco smoke during pregnancy (n = 55) either at home or at the workplace, and the other to mothers who were not routinely exposed to any tobacco smoke (n = 31). We excluded infants of women with conditions known to elevate nucleated RBC counts. There were no differences between groups in birth weight, maternal age, gravidity, parity, maternal analgesia during labor, 1- and 5-minute Apgar scores. Gestational age was minimally higher in the control group (39.6 +/- 1.1 vs 39.2 +/-.8 weeks). The median absolute nucleated RBC count in the passive smoking group was 357 x 10(6)/L (range: 0-5091 x 10(6)/L) versus 237 x 10(6)/L (range: 0-1733 x 10(6)/L) in nonsmoking controls. Stepwise regression analysis that included Apgar scores, gestational age, and the passive smoking status (yes/no) as independent variables showed significant correlation of absolute nucleated RBC count only with the passive smoking status. At birth, term AGA infants born to mothers exposed to passive smoking have increased circulating absolute nucleated RBC counts compared with those of controls. We speculate that passive smoking in pregnancy should be avoided, because it may have subtle negative effects on fetal oxygenation.