Benefits and Risks of Iron Supplementation in Anemic Neonatal Pigs

Iron deficiency is a common health problem. The most severe consequence of this disorder is iron deficiency anemia (IDA), which is considered the most common nutritional deficiency worldwide. Newborn piglets are an ideal model to explore the multifaceted etiology of IDA in mammals, as IDA is the most prevalent deficiency disorder throughout the early postnatal period in this species and frequently develops into a critical illness. Here, we report the very low expression of duodenal iron transporters in pigs during the first days of life. We postulate that this low expression level is why the iron demands of the piglet body are not met by iron absorption during this period. Interestingly, we found that a low level of duodenal divalent metal transporter 1 and ferroportin, two iron transporters located on the apical and basolateral membrane of duodenal absorptive enterocytes, respectively, correlates with abnormally high expression of hepcidin, despite the poor hepatic and overall iron status of these animals. Parenteral iron supplementation by a unique intramuscular administration of large amounts of iron dextran is current practice for the treatment of IDA in piglets. However, the potential toxicity of such supplemental iron implies the necessity for caution when applying this treatment. Here we demonstrate that a modified strategy for iron supplementation of newborn piglets with iron dextran improves the piglets’ hematological status, attenuates the induction of hepcidin expression, and minimizes the toxicity of the administered iron.Iron deficiency is the most frequent cause of anemia in humans,¹ and iron deficiency anemia (IDA) is most prevalent in the neonatal period and early childhood.² Early postnatal iron deficiency is also a widespread phenomenon in other mammalian species. Indeed, IDA has long been recognized as a serious iron disorder in livestock, especially in suckling piglets.³ In contrast to humans and mice, the complex molecular regulation of body iron homeostasis⁴ has been the subject of few studies in pigs.^5,6,7 Our knowledge of iron homeostasis in newborn pigs is mostly based on physiological concepts stemming from the early 1990s. Although the pig is a major biomedical mammalian model for human studies,⁸ its contribution to understanding the molecular pathophysiological mechanisms of human iron disorders has so far been small.⁹The common reason for iron deficiency in newborn piglets is their rapid growth, particularly the increase in blood volume and the number of red blood cells (RBCs). Indeed, erythroid precursors in the bone marrow use most of the iron found in the plasma for the synthesis of hemoglobin. As sow’s milk largely provides piglets with iron below their daily requirements,¹⁰ an exogenous source of iron is essential to prevent the reduction in RBC hemoglobin level. Intramuscular administration of large amounts of iron dextran (FeDex) on days 3 to 6 postpartum is current practice in the swine industry,^3,11 and has been proven to rectify the hematological status of piglets. However, it seems unlikely that 100 to 200 mg of iron (a commonly applied dose) given in a single injection to a piglet with only about 40 to 50 mg of iron in its body at birth,¹² is efficiently metabolized and detoxified. Moreover, high parenteral intake of supplemental iron may easily perturb the tight control of systemic iron metabolic processes. In this context, iron supplementation in piglets raises the question of the role of hepcidin (Hepc)—the systemic iron-regulatory hormone¹³ in the regulation of natural intestinal iron absorption, especially under conditions of morphological and functional rebuilding of the pig small intestinal mucosa that occurs within 21 days of birth.¹⁴In this study we examined the expression of proteins involved in iron absorption and its regulation in newborn piglets during the first 2 weeks of life. We also tested a modified strategy for iron supplementation of anemic piglets and demonstrated its ability to improve their hematological status, regulate Hepc expression and minimize the toxicity of supplemental iron.     

Successes and pitfalls of chronic peritoneal dialysis in infants – a Polish nationwide outcome study

Introduction

Peritoneal dialysis (PD) is a preferred method of renal replacement therapy for end-stage renal disease in children. Recent advances have allowed chronic PD to be provided to children of all ages and sizes.

Material and methods

The study was designed as a national (10 dialysis centres), multicentre retrospective analysis of the medical history of 33 children who started chronic peritoneal dialysis in their infancy between 1993 and 2005, with a follow-up period of at least 24 months.

Results

The nutritional status of the infants was unsatisfactory. The mean SDS of body weight at the start was –2.0, at 1 year of age –1.7. Only 40% of infants were adequately nourished at 1 year of age. Long-term follow-up analysis showed that 12 children received a kidney transplant, 13 were still on dialysis (4 changed method) and 6 died (mortality rate in the first year of life of 9%). In 2 children we observed an improvement of renal function. We observed a relatively high (1/8.8 patient-months) peritonitis rate in the analysed children when compared to 1 : 22 patient-months in all children undergoing PD in Poland.

Conclusions

The results of our survey have shown that the management of dialysed infants is still a challenge for the medical team and families, but long-term results of the therapy are encouraging.     

A novel founder CHEK2 mutation is associated with increased prostate cancer risk

Variants in the CHEK2 have been found to be associated with prostate cancer risk in the United States and Finland. We sequenced CHEK2 gene in 140 Polish patients with prostate cancer and then genotyped the three detected variants in a larger series of prostate cancer cases and controls. CHEK2 truncating mutations (IVS2 + 1G>A or 1100delC) were identified in 9 of 1921 controls (0.5%) and in 11 of 690 (1.6%) unselected patients with prostate cancer [odds ratio (OR) = 3.4; P = 0.004]. These mutations were found in 4 of 98 familial prostate cases (OR = 9.0; P = 0.0002). The missense variant I157T was also more frequent in men with prostate cancer (7.8%) than in controls (4.8%), but the relative risk was more modest (OR = 1.7; P = 0.03). I157T was identified in 16% of men with familial prostate cancer (OR = 3.8; P = 0.00002). Loss of the wild-type CHEK2 allele was not observed in any of prostate cancers from five men who carried CHEK2-truncating mutations. Our results provide evidence that the two truncating mutations of CHEK2 confer a moderate risk of prostate cancer in Polish men and that the missense change appears to confer a modest risk.     

Influence of home-based telemonitored Nordic walking training on autonomic nervous system balance in heart failure patients

Introduction

Rehabilitation positively affects the modulation of the autonomic nervous system (ANS). There are no papers evaluating the influence of Nordic walking training (NW) on ANS activity among chronic heart failure (CHF) patients. The aim of study was to assess the influence of NW on ANS activity measured by heart rate variability (HRV) and heart rate turbulence (HRT) in CHF patients and its correlation with physical capacity improvement measured by peak oxygen consumption (peak VO₂ [ml/kg/min]) in the cardiopulmonary exercise treadmill test (CPET).

Material and methods

The study group comprised 111 CHF patients (NYHA class II–III; ejection fraction (EF) ≤ 40%). Patients were randomized (2 : 1) to 8-week NW (five times weekly) at 40–70% of maximal heart rate (training group – TG) (n = 77), or to a control group (CG) (n = 34). The effectiveness of NW was assessed by changes (delta (Δ)) in peak VO₂, HRV and HRT as a result of comparing these parameters from the beginning and the end of the programme.

Results

Eventually, 36 TG patients and 15 CG patients were eligible for HRV and HRT analysis. In the TG low/high frequency ratio (LF/HF) decreased (1.9 ±1.11 vs. 1.7 ±0.63, p = 0.0001) and peak VO₂ increased (16.98 ±4.02 vs. 19.70 ±4.36 ml/kg/min, p < 0.0001). Favourable results in CG were not observed. The differences between TG and CG were significant: Δpeak VO₂ (p = 0.0081); ΔLF/HF (p = 0.0038). An inverse correlation was found between the decrease in ΔLF/HF and the increase in Δpeak VO₂ (R = –0.3830, p = 0.0211) only in the TG. Heart rate variability did not change significantly in either group.

Conclusions

Nordic walking positively affects the parasympathetic-sympathetic balance in CHF patients, which correlates with the improvement in Δpeak VO₂. No significant influence of NW on HRT was observed.     

Human papillomavirus infection predicts a better survival rate in patients with oropharyngeal cancer

IntroductionSquamous cell carcinoma is the most common malignant tumour occurring in the head and neck region. It is now understood that (human papillomavirus (HPV)- positive and HPV-negative diseases are two very different clinical entities associated with different outcomes. We decided to assess p16 expression status in patients with oropharyngeal cancer and retrospectively evaluate the outcomes of the treatment.Material and methodsThe evaluated group consisted of 98 consecutive patients with squamous cell carcinoma of the oropharynx treated in a combined way in Holycross Cancer Centre in Kielce in 2006–2014. For all patients p16 status was assessed based on the biological material. In 51 patients HPV infection was diagnosed. The Kaplan-Meier method was used to produce survival curves using the log-rank test and the Cox proportional hazard model was used to determine the risk factors. The following risk factors were included: HPV status (positive, negative), sex, age, smoking, histopathological grade of the tumour, clinical stage, and systemic therapy application. For HPV-positive and HPV-negative patients independent analyses were done including aforementioned factors, excluding HPV status.ResultsThe observation time for HPV-positive patients was significantly longer (p = 0.0008). Fifty-eight patients died, 40 patients are alive. Number of deaths in HPV-negative patients was statistically significantly higher (p = 0.0222). A statistically significant difference in the disease-free survival probability and overall survival probability between HPV-positive and HPV-negative patients was found (p = 0.0045 and p = 0.0037 respectively). For disease-free survival a statistically significant factor of the risk of recurrence was HPV infection (p = 0.0169). For HPV-positive patients, age (p = 0.0199) and smoking (p = 0.0353) were statistically significant risk factors of recurrence. For HPV-negative patients significant risk factors of recurrence were clinical stage (p = 0.0114) and systemic therapy application (p = 0.0271). For overall survival for the entire group statistically significant risk factors were absence of HPV infection (p = 0.0123), male sex (p = 0.0426), and age (p = 0.0311). For HPV-positive patients, age (p = 0.0096) and smoking (p = 0.0387) were statistically significant risk factors of death. For HPV-negative patients significant risk factors of death were clinical stage (p = 0.0120) and systemic therapy application (p = 0.0460).ConclusionsOur data show that HPV infection is a predictor of better disease-free and overall survival in patients with oropharyngeal cancer. For HPV-positive oropharyngeal cancer patients weekly given cisplatin with concurrent radiotherapy can be an alternative to three weekly given cisplatin considering effectiveness and early toxicity.     

Physical activity and angiotensin-converting enzyme gene polymorphism in mild hypertensives

It has been suggested that the insertion(I) allele of the I/deletion(D) polymorphism of the angiotensin converting enzyme (ACE) gene is associated with endurance exercise and increased physical conditioning in response to this type of exercise. To investigate the association between the ACE I/D polymorphism and physical activity status in 355 never treated, stage I hypertensives (265 men, 90 women, mean age: 33 +/- 9 years), in whom power exercise is contraindicated, participants of the HARVEST study. Physical activity was assessed using a standardized questionnaire. BMI and age did not vary among genotypes. None of active subjects performed power oriented exercises. ACE I/D frequencies (II-18%, ID-55%, DD-27%) were in Hardy-Weinberg equilibrium. Sedentary lifestyle was more common among DD than II hypertensives (76% in DD, and 48% in II, Chi(2) = 13.9, P = 0.001). In stepwise MANOVA using age, marital status, profession, sex, and ACE genotype as predictors of physical activity, marital status (F = 24.4, P < 0.0001) and ACE genotype (F = 16.03, P < 0.0001) contributed to more than 50% of the variance in physical activity status of the population. Our results suggest that the ACE I/D polymorphism may be a specific genetic factor associated with physical activity levels in free-living borderline and mild hypertensive subjects.     

Non-linear relationships between the BMI and physical fitness in Polish adolescents

Background: Comparisons of physical fitness in normal weight and overweight/obese youth generally highlight the negative consequences of an elevated BMI. In contrast, several studies of children and adolescents highlight the importance of variation in indicators of physical fitness across the full spectrum of BMIs from low through high.

Aim: The purpose of the study was to evaluate relationships between specific physical fitness items and the BMI among youth.

Subjects and methods: Height, weight and six physical fitness tests were measured in 1239 males and 903 females, aged 13–16?years; BMI (kg/m²) was calculated. Fitness tests were compared by weight status using sex-specific MANCOVAs, controlling for age. Sex-specific quadratic regressions of each fitness item on the BMI were also calculated.

Results: The sprint, standing long jump and shuttle run indicated better performances in normal than in thin and overweight/obese boys; the latter groups did not differ. Thin and normal weight boys performed better than overweight/obese boys in sit-ups and distance run. Among girls, sit-ups differed as follows: normal?>?thin?>?overweight/obese. Thin and normal weight girls performed better than overweight/obese girls in the jump, distance run and shuttle run. Normal weight girls were faster in the sprint than the overweight/obese. The quadratic regressions indicated significant non-linear relationships between the BMI and all fitness items among boys and in four items among girls.

Conclusion: Performances on fitness tests varied with weight status. Relationships between performances and the BMI were curvilinear except for the distance run and flexibility in girls.     

Evaluation of lymphocytes CD4+ and CD8+ and expression of ZAP-70 kinase on CD3+ and CD19+ lymphocytes in obese patients undergoing laparoscopic cholecystectomy

Background

Obesity has become a global epidemic and a leading metabolic disease in the world. Laparoscopic surgeries may influence the function of the immunologic system. The percentages of CD4+ and CD8+ T lymphocyte cells have been described as prognostic factors for patients undergoing abdominal surgeries. This study aimed to evaluate the changes in CD4+ and CD8+ T lymphocyte cells, the ratio of CD4+ to CD8+ cells, and the ZAP-70 kinase expression on T CD3+ and B CD19+ cells in obese and normal-weight individuals undergoing laparoscopic cholecystectomy (LC).

Methods

The study group consisted of 46 asymptomatic patients with gallstones shown by ultrasound examination but without signs of any gallbladder complications. The patients underwent planned LC. Blood samples were obtained at three times, and the percentages of studied cells were measured by flow cytometry. Patients were enrolled to two groups: N group (body mass index [BMI], ≤25 kg/m²) and O group (BMI, ≥30 kg/m²). For statistical analysis, the Mann–Whitney U test and the Wilcoxon matched-pairs signed-ranks test were used. All p values lower than 0.05 were considered significant.

Results

The percentage of CD4+ T cells did not differ between the N and O groups before or after the surgery. Only in the N group did the percentage of CD4+ lymphocytes increase from 0 to 48 h. A higher percentage of CD8+ lymphocytes was observed in the O group postoperatively than in the N group. Differences of ZAP-70 kinase expression in the O group were observed at 24 and 48 h of the study. Decreased expression of ZAP-70 kinase was shown in the N group at both 0–24 and 24–48 h. In the O group, this tendency was noted at 24–48 h.

Conclusions

Immunologic activation after LC was confirmed in both weight groups. However, higher modulation, more typical for open surgeries, was observed in the obese group.     

Multigene adenoviral therapy for the attenuation of ischemia-reperfusion injury after preservation for cardiac transplantation

OBJECTIVE: The protective effect of adenovirus-mediated ex vivo multigene transfer with superoxide dismutase, a free radical scavenger, and nitric oxide, a vasodilator with anti-inflammatory properties, was examined in the rat heart during experimental ischemia-reperfusion mimicking preservation for cardiac transplantation. METHODS: Donor rat hearts (n = 6 per group) were perfused with solution containing adenoviral vector carrying genes for beta-galactosidase (group A), endothelial nitric oxide synthase (group B), manganese superoxide dismutase (group C), or both endothelial nitric oxide synthase and manganese superoxide dismutase (group D). Hearts were then implanted heterotopically into the abdomens of recipient rats. Four days later, transplanted hearts were collected, connected to a Langendorff perfusion apparatus, and subjected to 6 hours of ischemia followed by 1 hour of reperfusion. Cardiac function was evaluated with an intraventricular balloon at the beginning of Langendorff perfusion and after ischemia-reperfusion. RESULTS: Effective gene transfection was confirmed with X-gal staining in group A hearts. Positive immunoreactivity for endothelial nitric oxide synthase, manganese superoxide dismutase, or both was present predominantly in cardiomyocytes in group B, C, and D hearts. Percentage recovery of preischemic left ventricular developed pressure was 62.1% +/- 7.36% in group A; recoveries were increased to 79.6% +/- 6.4%, 86.8% +/- 9.1%, and 79.4% +/- 6.2% in groups B, C, and D, respectively. CONCLUSION: These results indicate that adenoviral gene transfer of manganese superoxide dismutase and endothelial nitric oxide synthase can attenuate myocardial ischemia-reperfusion injury, with the former providing the most significant protection. Combined overexpression of manganese superoxide dismutase and endothelial nitric oxide synthase did not enhance myocardial recovery any further.