Theoretical and Computational Analysis of the Thermal Quasi-Geostrophic Model.

This work involves theoretical and numerical analysis of the thermal quasi-geostrophic (TQG) model of submesoscale geophysical fluid dynamics (GFD). Physically, the TQG model involves thermal geostrophic balance, in which the Rossby number, the Froude number and the stratification parameter are all of the same asymptotic order. The main analytical contribution of this paper is to construct local-in-time unique strong solutions for the TQG model. For this, we show that solutions of its regularised version α-TQG converge to solutions of TQG as its smoothing parameter α→0 and we obtain blow-up criteria for the α-TQG model. The main contribution of the computational analysis is to verify the rate of convergence of α-TQG solutions to TQG solutions as α→0, for example, simulations in appropriate GFD regimes.

The potential effects and interactions of oxidative stress and trace minerals on fresh and frozen semen in bulls - a review.

Reproduction is one of the most important factors determining successful cattle farming systems. Management practices, such as nutritional supplementation, can influence the reproductive performance of cattle. The objective of this literature review is to determine the potential value of injectable trace mineral administration on fresh and cryopreserved semen quality of bulls. A search of keywords related to the topic was performed on published articles and textbooks. The search was narrowed to the 40 most relevant references. Several studies have demonstrated a positive association between trace mineral supplementation and bull semen quality. Moderate amounts of reactive oxygen species (ROS) play an important role in normal spermatogenesis, but oxidative stress (OS), as experienced with adverse environmental conditions or disease, can contribute to idiopathic male infertility by negatively impacting spermatogenesis. Trace minerals such as selenium, copper, zinc, and manganese have been demonstrated to have antioxidant effects in mammals. Due to the complexity of oral ingested trace mineral bioavailability, injectable trace mineral supplementation prior to physiological periods with known deficiencies or increased requirement can benefit the animal. The potential benefits of injectable trace mineral supplementation to minimise oxidative damage to spermatogenesis in breeding bulls need further investigation. Positive results from such studies can lead to the implementation of injectable trace mineral supplementation strategies prior to the breeding season to minimise the detrimental effects of OS and can improve semen quality.

Oocyte quality and viability in Nguni and Hereford cows exposed to different levels of dietary protein.

High blood urea nitrogen (BUN) concentration decreases fertility in ruminants. Nguni cattle are reported to maintain BUN concentration more efficiently than other beef breeds. Our objectives were to determine if BUN concentration differed between Nguni and Hereford cows exposed to a high protein ration, and if breed or BUN and serum protein concentrations at the time of oocyte pick-up affected oocyte quantity, quality, and viability. Twelve Nguni and 10 Hereford cows were randomized into high or normal BUN-inducing diets in a crossover design. Ultrasound-guided oocyte pick-up was performed twice weekly; oocytes were counted, visually graded and the viable oocytes were pooled by treatment and breed for in vitro maturation, fertilization, and culture. Nguni cows on the highest protein ration achieved lower mean BUN concentration than Herefords (P < 0.05), and Nguni cows reached BUN concentrations above 20 mg/dL less frequently than Herefords (P = 0.03). Donor BUN concentration above 20 mg/dL at the time of oocyte pick-up, but not breed, independently decreased the number of good quality oocytes harvested. Increasing weighted mean serum albumin of donor cows was independently associated with the number of oocytes that cleaved by day 2 and that reached morula stage by day 7 (P = 0.01). In conclusion, Nguni cows reached the critical threshold of 20 mg/dL BUN less frequently than Herefords; BUN of donor cows above 20 mg/dL negatively affected visual oocyte quality independent of breed, and increasing serum albumin of donor cows improved viability of bovine oocytes.

Dynamics of non-holonomic systems with stochastic transport.

This paper formulates a variational approach for treating observational uncertainty and/or computational model errors as stochastic transport in dynamical systems governed by action principles under non-holonomic constraints. For this purpose, we derive, analyse and numerically study the example of an unbalanced spherical ball rolling under gravity along a stochastic path. Our approach uses the Hamilton-Pontryagin variational principle, constrained by a stochastic rolling condition, which we show is equivalent to the corresponding stochastic Lagrange-d'Alembert principle. In the example of the rolling ball, the stochasticity represents uncertainty in the observation and/or error in the computational simulation of the angular velocity of rolling. The influence of the stochasticity on the deterministically conserved quantities is investigated both analytically and numerically. Our approach applies to a wide variety of stochastic, non-holonomically constrained systems, because it preserves the mathematical properties inherited from the variational principle.

Stochastic partial differential fluid equations as a diffusive limit of deterministic Lagrangian multi-time dynamics.

In Holm (Holm 2015 Proc. R. Soc. A471, 20140963. (doi:10.1098/rspa.2014.0963)), stochastic fluid equations were derived by employing a variational principle with an assumed stochastic Lagrangian particle dynamics. Here we show that the same stochastic Lagrangian dynamics naturally arises in a multi-scale decomposition of the deterministic Lagrangian flow map into a slow large-scale mean and a rapidly fluctuating small-scale map. We employ homogenization theory to derive effective slow stochastic particle dynamics for the resolved mean part, thereby obtaining stochastic fluid partial equations in the Eulerian formulation. To justify the application of rigorous homogenization theory, we assume mildly chaotic fast small-scale dynamics, as well as a centring condition. The latter requires that the mean of the fluctuating deviations is small, when pulled back to the mean flow.

Macroscopic and microscopic evaluation of Eurasian lynx (Lynx lynx) female tubular reproductive organs in relation to ovarian structures.

Although monitoring wild animals in the field is essential for estimations of population size and development, there are pitfalls associated with field monitoring. In addition, some detailed data about reproductive physiology can be difficult to obtain in wild live animals. Studying reproductive organs from the Eurasian lynx killed at hunting or found dead could be used as a valuable addition to other field data. We evaluated reproductive organs from 39 Eurasian lynx females (Lynx lynx) killed in Sweden during the hunting seasons in 2009, 2010, and 2011. According to notes on ovarian structures, the animals were categorized as being in one of four different reproductive stages: juvenile (n = 10), follicular stage (n = 8), luteal stage (n = 11), and anestrus (n = 10). Corpora lutea were classified as fresh CL from the present season or as luteal bodies from previous cycles. Microscopic evaluations were blindly coded while the outer measurements of the vagina and uterus were taken at the time of organ retrieval. The width of the endometrium, myometrium, outer width of the uterine horns, and the diameter of the vagina differed significantly with the reproductive stage (P < 0.001) and were largest in the follicular and luteal phases. The number of endometrial glands evaluated blindly coded on a subjective scale was significantly associated with the reproductive stage (P < 0.0001) and was significantly higher in the luteal phase than that in any other reproductive stages (P < 0.05). Cornification of the vaginal epithelium was only observed in females in the follicular stage or in females with signs of a recent ovulation. In conclusion, both macroscopic and histologic measurements are useful for a correct classification of the reproductive stage when evaluating reproductive organs in the Eurasian lynx killed during the hunting season. Routine evaluation of reproductive organs has a potential to be a useful additional tool to field studies of live lynx to monitor their reproduction.

Effects of broiler breeder management on pullet body weight and carcass uniformity.

An experiment was conducted to study the effect of broiler breeder feeding management practices on pullet performance, BW uniformity, and carcass traits during rearing (to 22 wk of age). At 3 wk of age, 1,200 Ross 308 breeder pullets were assigned to one of 5 treatments: 1) control: standard mash diet, fed daily; 2) high fiber: mash diet containing 25% lower nutrient density, fed daily; 3) scatter: standard diet in pellet form scattered on litter, fed daily; 4) skip-a-day: standard mash diet, fed on alternate days; or 5) grading: standard mash diet, fed daily (birds sorted into low, average, and high BW groups every 4 wk). Birds on the high fiber treatment consumed more feed (P<0.0001) and had the highest feed conversion ratio (FCR; P<0.004) but the lowest ME to gain and CP to gain ratios (P≤0.002). Skip-a-day treatment pullets consumed more ME and CP than birds in any other treatment (P<0.001). Grading yielded the highest BW uniformity at 22 wk of age (CV=6.2%), while control and high fiber treatment groups were least uniform (CV>15%; P<0.0001). Skip-a-day feed restriction produced birds with the significantly lowest breast muscle and highest liver weight compared to all other treatments (P<0.05). Variation in shank length, chest width, and breast muscle was lowest in the grading treatment, whereas the CV for fat pad and liver was lowest in the skip-a-day treatment. In this trial, broiler breeder target BW profiles were achieved using combinations of quantitative and qualitative feed restriction, or preemptive management practices. Qualitative diet dilution and skip-a-day management did little to increase flock uniformity relative to the control during the most intense period of feed restriction (7 to 19 wk). Scatter feeding increased flock uniformity to a small degree, whereas grading yielded the highest increase in BW and carcass trait uniformity.

Evaluation of pre-breeding reproductive tract scoring as a predictor of long term reproductive performance in beef heifers.

In a 7-year longitudinal study 292 Bovelder beef cows in a restricted breeding system in South Africa were observed from 1 to 2 days before their first breeding season, when reproductive tract scoring (RTS, scored from 1 to 5) was performed, until weaning their 5th calves. The objective was to determine whether pre-breeding RTS in heifers is a valid tool to predict long-term reproductive performance. Outcomes measured were failure to show oestrus during the first 24 days of the first 50-day AI season (24-day anoestrus), failure to become pregnant during each yearly artificial insemination (AI) season (reproductive failure), number of days from the start of each AI season to calving, and number of years to reproductive failure. The effect of RTS on each outcome was adjusted for year of birth, pre-breeding age, BW and body condition score (BCS), and for 24-day anoestrus, bull, gestation length, previous days to calving and previous cow efficiency index, the latter two in the case of the 2nd to the 5th calving season. During their first breeding season, heifers with RTS 1 and 2 combined were more likely to be in anoestrus for the first 24 days (OR=3.0, 95% CI 1.5, 6.4, P=0.003), and were also more likely to fail to become pregnant even after adjusting for 24-day anoestrus (OR=2.1, 95% CI 1.1, 3.9, P=0.025), compared to those with RTS 4 and 5 combined. Animals with RTS 1 and 2 combined were at increased risk of early reproductive failure compared to those with RTS 4 and 5 combined (HR=1.4, 95% CI 1.0, 1.9, P=0.045) although RTS was not associated with calving rate or days to calving after the second calving season. Low RTS at a threshold of 1 had consistent specificity of ≥94% for both 24-day anoestrus and pregnancy failure, however its predictive value was lower in the age cohort with a higher prevalence of anoestrus. We conclude that RTS is a valid management tool for culling decisions intended to improve long-term reproductive success in a seasonal breeding system, by excluding heifers that are likely to fail to become pregnant or likely to calve late during their first calving season.

Optimization of in vitro inhibition of HT-29 colon cancer cell cultures by Solanum tuberosum L. extracts.

Secondary metabolites in potato have been reported to possess bioactive properties, including growth inhibition of cancer cells. Because potatoes are widely consumed globally, potential health benefits may have broad application. Thus we investigated growth inhibition of HT-29 colon cancer cell cultures by extracts from 13 diverse genetic breeding clones. Extracts from three pigmented selections (CO97226-2R/R, CO97216-1P/P, CO04058-3RW/RW) inhibited growth of in vitro HT-29 cell cultures more effectively than other clones tested. While inhibition was highest from pigmented selections and pigmented tuber tissue sectors, not all pigmented breeding lines tested had appreciable inhibitory properties. Thus, inhibition was not uniquely linked to pigmentation. Immature tubers had the highest inhibitory properties, and in most cases mature tubers retained very low inhibition properties. Flowers and skins inhibited strongly at lower extract concentrations. An extract consisting of 7.2 mg mL⁻¹ cell culture medium was the lowest effective concentration. While raw tuber extracts inhibited most effectively, a few clones at higher concentrations retained inhibition after cooking. Heated whole tubers retained higher inhibition than heated aqueous extracts. While all aqueous extracts from the two tuber selections (CO97216-1P/P and CO97226-2R/R) inhibited HT-29 cell cultures, inhibition was significantly enhanced in purple pigmented tubers of CO97216-1P/P prepared cryogenically as liquid nitrogen powders compared to extracts from freeze dried samples. Upregulation of caspase-3 protease activity, indicative of apoptosis, was highest among the most inhibitory clone samples. The unique sectorial red pigment expressing selection (CO04058-3RW/RW) provided a model system that isolated expression in pigmented sectors, and thus eliminated developmental, environmental and genetic confounding.

A new application of pelvis area data as culling tool to aid in the management of dystocia in heifers.

Although fetomaternal disproportion is the major cause of dystocia in heifers, pelvis area (PA) is not recommended as a culling tool due to its relatively low importance and genetic correlation with calf birth weight (BWT), the most important factor associated with dystocia. The objective of this observational study of 484 limited bred yearling beef heifers was to compare the effects of different methods of adjustment of PA data for culling to select against dystocia. Multivariable analyses were used to determine predictors of PA, calf BWT, and dystocia. Hypothetical culling rates of 10 and 20% were then applied after ranking heifers by each of the following: unadjusted PA, PA adjusted to 365 d of age by subtracting 0.27 cm(2) per day of age difference between each heifer's age and 365 d (APA), PA:prebreeding BW ratio (PA:BW), PA adjusted to the median BW of the group using the regression coefficient of PA on BW within age group (BWPA), and PA similarly adjusted to the median lean BW (LBWPA). Dam parity, sire, prebreeding age, prebreeding BW, and prebreeding BCS were associated with PA whereas dam parity, sire, own BWT, PA, AI bull, and calf gender were associated with calf BWT (P < 0.05). Dam parity, calf BWT, and either BWPA or LBWPA were the only independent predictors of dystocia (P < 0.05). Adjusting PA to BW or lean BW (LBW) improved the sensitivity and specificity to predict dystocia. After hypothetical culling by PA, retained heifers were heavier and had a higher calving rate and calves tended to be heavier at birth compared to culled heifers, but dystocia rates were not different. Culling by APA resulted in similar effects, except that dystocia rate tended to be lower in retained heifers. Culling by PA:BW resulted in lower dystocia rate in retained than in culled heifers, but retained heifers had lower prebreeding BW than culls. Culling by BWPA and LBWPA resulted in lower proportions with dystocia and a tendency towards higher calving rates in the retained heifers, without affecting the prebreeding BW or calf BWT. It is concluded that pelvimetry is a useful culling tool to aid in the management of dystocia in yearling heifers and that adjustment of PA to median BW or LBW within age group improves its accuracy and avoids the undesirable side effects.

Seminal transmission of lumpy skin disease virus in heifers.

It is known that lumpy skin disease virus (LSDV) can be shed in bull semen following infection and also that artificial insemination (AI) poses a biosecurity risk. However, it is not known whether the use of LSDV infected semen in AI poses a biosecurity risk. The aim of this study was to investigate whether LSDV, transmitted through semen, can infect cows and their embryos. Two controlled trials were performed simultaneously. Eleven young beef heifers, naïve to LSDV, were synchronized using an OvSynch protocol and inseminated on Day 0 with fresh semen spiked with a field strain of LSDV on day 0. Six of the heifers were superovulated on Day 1 using pregnant mare serum gonadotropin, and embryos were flushed from these heifers on Day 6. Blood and serum samples were collected from Day 4 until Day 27 to determine the presence of LSDV by PCR and virus isolation, and the presence of antibodies against LSDV by SNT. The first clinical signs of LSD were noticed on Day 10, followed by severe generalized LSD in three heifers and mild LSD in two more heifers. Two heifers were humanely euthanized due to severe unresponsive stranguria. LSDV was detected by PCR, virus isolation or electron microscopy in blood, embryos and organs of experimentally infected animals; and eight heifers had seroconverted by Day 27. Two control animals were not affected. This is the first report of experimental seminal transmission of LSDV in cattle.

Collection, processing and testing of bone, corneas, umbilical cord blood and haematopoietic stem cells by European Blood Alliance members.

BACKGROUND AND OBJECTIVES: A questionnaire study was carried out in collaboration with the European Blood Alliance (EBA) Tissues and Cells (T&C) working group. The aim was to assess the level of involvement and commonality of processes on the procurement, testing and storage of bone, corneas, umbilical cord blood (UCB) and haematopoietic stem cells (HSC) in order to identify different practices and to explore whether recommendations can be made for harmonization. MATERIALS AND METHODS: An online questionnaire was used for data collection in 2011, and 43 replies were received covering 71 product answers from 13 countries. RESULTS AND CONCLUSIONS: Estimated percentages of tissue and cell banking covered by EBA member blood banks as a proportion of all collections of each individual country varied markedly. There were also major differences in the amounts of products collected and discarded and in proportions tissues provided for grafting. However, discarding of certain collections also reflects the practice of increasing the likelihood of the very best units being used for transplantation. Harmonization of possible practices should focus on matching supply with demand and on identifying the most efficient operators. This could allow for the development of practices for minimizing unnecessary collections.

Quasiconservation laws for compressible three-dimensional Navier-Stokes flow.

We formulate the quasi-Lagrangian fluid transport dynamics of mass density ρ and the projection q=ω·∇ρ of the vorticity ω onto the density gradient, as determined by the three-dimensional compressible Navier-Stokes equations for an ideal gas, although the results apply for an arbitrary equation of state. It turns out that the quasi-Lagrangian transport of q cannot cross a level set of ρ. That is, in this formulation, level sets of ρ (isopycnals) are impermeable to the transport of the projection q.

Left-right difference in fetal liver oxygenation during hypoxia estimated by BOLD MRI in a fetal sheep model.

OBJECTIVE: The purpose of this study was to measure differences in oxygenation between the left and right sides of the fetal liver during varying oxygenation levels. METHODS: Eight ewes carrying singleton fetuses at gestational age 125 days (term, 145 days) were included in the study. Under anesthesia the ewes were ventilated with gas containing different levels of oxygen, thereby subjecting the fetuses to hyperoxia (mean ± SD maternal arterial partial pressure of oxygen (pO2), 23.2 ± 8.2 kPa) and hypoxia (mean maternal arterial pO2, 7.1 ± 0.5 kPa). Changes in oxygenation within the fetal liver were assessed by blood oxygen level-dependent (BOLD) magnetic resonance imaging (MRI). RESULTS: During hyperoxia there was no difference between the BOLD signal in the left and right sides of the fetal liver; mean change in BOLD (ΔBOLD)(hyperox), -0.9 ± 3.7%. During hypoxia, however, the decrease in the BOLD signal was more pronounced in the right side as compared with the left side, thereby creating a significant increase in the left-right difference in the BOLD signal; mean ΔBOLD(hypox), 5.2 ± 2.2% (P = 0.002, paired t-test). The left-right difference was directly proportional to the degree of hypoxia (R2 = 0.86, P = 0.007). CONCLUSIONS: To our knowledge, this is the first study demonstrating differences in oxygenation between the left and right sides of the fetal liver during hypoxia, a difference that can be explained by increased ductus venosus shunting. Thus, the BOLD MRI technique is a promising non-invasive tool that might be useful for the future monitoring of the human fetus.

Extreme events in solutions of hydrostatic and non-hydrostatic climate models.

Initially, this paper reviews the mathematical issues surrounding hydrostatic primitive equations (HPEs) and non-hydrostatic primitive equations (NPEs) that have been used extensively in numerical weather prediction and climate modelling. A new impetus has been provided by a recent proof of the existence and uniqueness of solutions of viscous HPEs on a cylinder with Neumann-like boundary conditions on the top and bottom. In contrast, the regularity of solutions of NPEs remains an open question. With this HPE regularity result in mind, the second issue examined in this paper is whether extreme events are allowed to arise spontaneously in their solutions. Such events could include, for example, the sudden appearance and disappearance of locally intense fronts that do not involve deep convection. Analytical methods are used to show that for viscous HPEs, the creation of small-scale structures is allowed locally in space and time at sizes that scale inversely with the Reynolds number.

Fatal congenital anaplasmosis associated with bovine viral diarrhoea virus (BVDV) infection in a crossbred calf.

Clinical disease resulting from the vertical transmission of Anaplasma marginale has only been reported on 5 occasions despite studies demonstrating successful in utero transmission. During the reported experimental induction of congenital anaplasmosis in calves, the outcome was variable but mostly led to inapparent or mild infection. There are previous case reports of fatal congenital anaplasmosis following natural infection. The clinical findings in a 2-day-old calf presented to the Onderstepoort Veterinary Academic Hospital with clinical signs of congenital anaplasmosis, which was unresponsive to treatment, are described. Subsequent post mortem diagnostic tests revealed that this calf was co-infected with bovine viral diarrhoea virus (BVDV). It is postulated that immunosuppression resulting from BVDV infection predisposed to severe, fatal anaplasmosis in this calf.

The value of reproductive tract scoring as a predictor of fertility and production outcomes in beef heifers.

In this study, 272 beef heifers were studied from just before their first breeding season (October 15, 2003), through their second breeding season, and until just after they had weaned their first calves in March, 2005. This study was performed concurrently with another study testing the economic effects of an estrous synchronization protocol using PG. Reproductive tract scoring (RTS) by rectal palpation was performed on the group of heifers 1 d before the onset of their first breeding season. The effect of RTS on several fertility and production outcomes was tested, and the association of RTS with the outcomes was compared with that of other input variables such as BW, age, BCS, and Kleiber ratio using multiple or univariable linear, logistic, or Cox regression. Area under the curve for receiver operating characteristic analysis was used to compare the ability of different input variables to predict pregnancy outcome. After adjustment for BW and age, RTS was positively associated with pregnancy rate to the 50-d AI season (P < 0.01), calf weaning weight (r = 0.22, P < 0.01), and pregnancy rate to the subsequent breeding season (P < 0.01), and negatively associated with days to calving (r = 0.28, P < 0.01). Reproductive tract scoring was a better predictor of fertility than was Kleiber ratio and similar in its prediction of calf weaning weight. It was concluded from this study that RTS is a predictor of heifer fertility, compares well with other traits used as a predictor of production outcomes, and is likely to be a good predictor of lifetime production of the cow.

The economic effects of an estrus synchronization protocol using prostaglandin in beef heifers.

We estimated the effect of estrus synchronization on reproduction, production and economic outcomes in 272 beef heifers randomly allocated to a synchronized Test group or an unsynchronized Control group. The Test group received AI upon estrus detection for 6 days followed by PGF2 treatment of heifers that had not shown estrus by day 6 (PGF/6). In both groups AI was continued for 50 days, followed by a 42-day bull breeding period. Heifers were followed through their second breeding season and until they had weaned their first calves. Synchronization resulted in a reduction in median days to first insemination (8 vs. 11 in the Test and Control groups, respectively, P<0.01) and median days to calving of calves born to AI (14 vs. 20, P=0.04). There was no significant difference in pregnancy rate to the AI period (60.0% vs. 51.8%, P=0.18), final pregnancy rate (82.2% vs. 83.2%, P=0.87) or pregnancy rate to the subsequent breeding season (96.0% vs. 95.0%, P=1.00). Although mean calf weaning mass was not significantly different (207.0 kg vs. 201.4 kg, P=0.32), the total mass of calves weaned in this study was 14,843 kg vs. 13,060 kg and the benefit: cost ratio for synchronization was 2.8. It was therefore concluded that a PGF/6 protocol may affect the total mass of calves weaned by changing days to calving, weaning rate, the ratio of male: female calves born and/or the birth mass of calves.

The occurrence of dermatosparaxis in a commercial Drakensberger cattle herd in South Africa.

Dermatosparaxis is a heritable collagen dysplasia causing skin extensibility and fragility. In Belgian Blue cattle this mutation has been described as a 3 base pair (bp) change followed by a 17bp deletion in the gene coding for procollagen 1 N-Proteinase (pNPI). An outbreak in a commercial Drakensberger herd in South Africa followed the introduction in late 2000 of a 3-year-old bull that developed skin lesions in 2001 and was culled in 2002. Some of his offspring were similarly affected, 1 of which was kept as a breeding bull after his sire's death. Two affected calves were referred to the Onderstepoort Veterinary Academic Hospital in October 2005. Detailed examination revealed only skin abnormalities limited to the lateral extremities of the thorax, abdomen and pelvis, viz. either acute lacerations of varying sizes, slow healing defects or thin scars in chronic cases. During a subsequent farm visit, 13 animals with similar wounds were seen in the herd of 146 animals. Electron microscopic examination of skin biopsies revealed haphazard arrangement and loose packing of dermal collagen fibrils within collagen fibres. The fibrils showed size variation and slightly irregular outlines on cross-section, consistent with mild dermatosparaxis. DNA samples of affected calves were analysed using primers designed to amplify the region of the pNPI gene that contained the mutation described in Belgian Blue cattle, but this mutation could not be demonstrated in any of the animals tested. It is concluded that a form of dermatosparaxis with a different gene mutation from that described in Belgian Blue cattle exists in Drakensberger cattle in South Africa. This possibly also explains the milder and more delayed clinical signs and the milder dermal collagen ultrastructural abnormalities.