Witold Krupski

Effects of combined maternal administration with alpha-ketoglutarate (AKG) and β-hydroxy-β-methylbutyrate (HMB) on prenatal programming of skeletal properties in the offspring

BackgroundNutritional manipulations during fetal growth may induce long-term metabolic effects in postnatal life. The aim of the study was to test whether combined treatment of pregnant sows with alpha-ketoglutarate and β-hydroxy-β-methylbutyrate induces additive long-term effects on skeletal system properties in the offspring.MethodsThe study was performed on 290 pigs obtained from 24 sows divided into 4 equal groups and subjected to experimental treatment during two weeks before delivery. The first group consisted of control sows, while the second group received alpha-ketoglutarate. The third group was treated with β-hydroxy-β-methylbutyrate and the fourth group underwent combined administration of alpha-ketoglutarate and β-hydroxy-β-methylbutyrate. Piglets obtained from sows were reared until slaughter age to perform morphometric, densitometric and mechanical analyses of femur. Serum evaluations of growth hormone, insulin-like growth factor-1, bone-specific alkaline phosphatase and osteocalcin were performed in newborns and 90-day old piglets; additionally, plasma amino acid concentration was measured in newborns.ResultsMaternal treatment with alpha-ketoglutarate and β-hydroxy-β-methylbutyrate significantly reduced fattening time and increased birth body weight, daily body weight gain, bone weight, volumetric bone mineral density, geometrical parameters and mechanical endurance of femur. These effects were associated with increased serum concentrations of growth hormone, insulin-like growth factor-1, bone-specific alkaline phosphatase and osteocalcin. Furthermore, alpha-ketoglutarate and β-hydroxy-β-methylbutyrate administered solely or in combination significantly increased plasma level of 19 amino acids.ConclusionsHormonal and amino acid evaluations in pigs indicate additive effects of AKG and HMB on systemic growth and development; however, determination of bone properties has not shown such phenomenon.

3-Hydroxy-3-methylbutyrate administration diminishes fundectomy-induced osteopenia of the lumbar spine in pigs

OBJECTIVE The aim of the study was to test the hypothesis that oral administration with 3-hydroxy-3-methylbutyrate (HMB) positively influences bone metabolism and diminishes fundectomy-induced osteopenia of the axial skeleton in pigs. The pig model was chosen because of its recognized physiologic and anatomic similarities of the gastrointestinal tract and skeletal system to those of humans. METHODS Eighteen male pigs were divided into three weight-matched groups at 40 d of life. Animals from the first and second groups were subjected to experimental fundectomy and the third group was sham operated. Starting the day after the fundectomy, the first and second groups received placebo and HMB, respectively. Animals were sacrificed at the age of 8 mo to obtain L(5) and L(6) vertebrae for analysis. The effects of HMB administration on plasma amino acids concentrations, bone mineral density, morphology, and mechanical properties of the lumbar vertebrae were determined. RESULTS The HMB treatment increased the weight of the vertebrae, bone mineral density, bone mineral content, total bone volume, trabecular bone mineral density, mean volumetric bone mineral density, calcium hydroxyapatite density in the trabecular and cortical bones, and plasma amino acid concentrations in the fundectomized pigs (P < 0.05). Mechanical strength of the spine, expressed by the values of ultimate force, Youngs modulus, ultimate stress, stiffness, and work to the ultimate force point was increased in HMB-treated pigs (P < 0.05). CONCLUSION HMB administration to fundectomized pigs improved plasma amino acids concentrations and diminished development of fundectomy-induced osteopenia of the axial skeleton.

Effects of dietary calcium content and vitamin D source on skeletal properties in growing turkeys

1. The aim of the study was to investigate the effects of feeding fast growing turkeys with differentiated dietary calcium (Ca) content, and the partial replacement of vitamin D3 in the feed with 25-hydroxycholecalciferol (25(OH)D3), on skeletal properties. 2. One-day-old Big-6 male turkeys (n = 1008) were randomly divided into 4 groups, and two subgroups were created within each group. The groups were differentiated with 4 levels of Ca provision in the feed, namely 85% of the National Research Council (NRC) recommendation (Group Ca1); 95% as above (Group Ca2); 105% as above (Group Ca3); and 115% as above (Group Ca4). The first subgroup received the recommended dosage of cholecalciferol (vitamin D3 subgroup) in the feed, while in the second subgroup (Hy-D subgroup), half of the dosage of cholecalciferol was replaced with 25(OH)D3. At the ages of 4, 8, 12 and 20 weeks, 7 turkeys from each subgroup were randomly selected and killed to obtain the right tibia for densitometric, geometric and mechanical analyses. 3. This study showed advantageous effects of increased calcium supply in the diet on skeletal system properties, that were increased and produced the most desirable traits in turkeys receiving 95%, 105% and 115% of the NRC calcium recommendation. Benefits resulting from administration of 25(OH)D3 in the diet were also obtained in the skeletal formation of turkeys, and the most advantageous effects were present in the group receiving 105% of recommended dietary Ca. 4. Effects on the metabolic response of the skeleton of turkeys to manipulation of dietary calcium content and vitamin D3 source were the most evident in the groups between 4 and 12 weeks of life, and demonstrated a limited ability to induce a positive influence on bone properties at advanced stages of the production cycle by alteration of these dietary factors.

Anatomical classification of the shape and topography of the stomach

The aim of the study was to present the classification of anatomical variations of the stomach, based on the radiological and historical data. In years 2006–2010, 2,034 examinations of the upper digestive tract were performed. Normal stomach anatomy or different variations of the organ shape and/or topography without any organic radiologically detectable gastric lesions were revealed in 568 and 821 cases, respectively. Five primary groups were established: abnormal position along longitudinal (I) and horizontal axis (II), as well as abnormal shape (III) and stomach connections (IV) or mixed forms (V). The first group contains abnormalities most commonly observed among examined patients such as stomach rotation and translocation to the chest cavity, including sliding, paraesophageal, mixed-form and upside-down hiatal diaphragmatic hernias, as well as short esophagus, and the other diaphragmatic hernias, that were not found in the evaluated population. The second group includes the stomach cascade. The third and fourth groups comprise developmental variations and organ malformations that were not observed in evaluated patients. The last group (V) encloses mixed forms that connect two or more previous variations.

The long‐term effect of α‐ketoglutarate, given early in postnatal life, on both growth and various bone parameters in pigs

The long-term effect of alpha-ketoglutarate (AKG) given for 21-24 days post-partum, on the skeleton of commercial pigs, was investigated. In experiment A, 12 pigs were given AKG [0.1 g/kg of body weight (b.w.) per day per os], while 12 controls were administered vehicle. At day 169, the left and right femur, humerus and sixth ribs were analysed for mechanical and geometrical properties and quantitative computed tomography. In experiment B, 32 piglets were divided equally into an AKG group (0.3 g/kg of b.w. per day) or a control group. Blood, taken at days 24 and 53 was analysed for plasma 17 beta-oestradiol. The main bone effect of AKG was to increase bone length in the sixth rib (7.3%, p < 0.01), ultimate strength (23%, p < 0.05), Young s modulus (52%, p < 0.001) and maximum elastic strength (31%, p = 0.056) compared with controls. In both experiments, AKG preferentially increased the growth of female piglets, whilst for male piglets AKG had the opposite effect. In addition, AKG elevated plasma 17 beta-oestradiol levels compared to those of controls at the end of the period of treatment (20%, p = 0.002). It is concluded that AKG has long-term effects on rib properties when given early in postnatal life whilst it elevates plasma 17 beta-oestradiol levels only so long as it is being administered.

Humoral response against glial derived antigens in Parkinson's disease

To check whether glial cells have the ability to elicit adaptive immune response in Parkinsons disease and whether a change in this immune response can be observed over time. There is an increasing evidence that glial cells are involved in the neurodegenerative process in PD, in addition to neuronal structures. Measurement of autoantibodies against proteins of oligodendrocytes may serve as an indirect method to assess the level of glial cells activation or degeneration under in vivo conditions. Serum samples from 26 PD patients were collected twice, at baseline and after mean of 13 months. In addition, serum samples from 13 healthy controls matched for age and gender were assessed at one time point. IgG and IgM autoantibodies against myelin-oligodendrocyticglycoprotein (MOG), myelin basic protein (MBP), myelin-associated glycoprotein (MAG) and proteolipoprotein (PLP) were measured in all investigated subjects by a commercially available ELISA system (Mediagnost, Germany). In a group of PD significant decrease of IgG titers was observed for anti-MAG autoantibodies over the investigated time period (p<0.05). For IgM antibodies, we observed statistically significant decrease in anti-MAG autoantibodies in the follow-up period (p<0.05) and increase in anti-MBP and anti-PLP autoantibodies (p<0.05). All antibody titers differed significantly between healthy control subjects and PD patients. Our study provides the evidence for the presence of humoral response against some glial derived antigens in PD. The increasing levels of anti MBP IgG and IgM might point to the value of this marker for monitoring disease progression.

Fundectomy-Evoked Osteopenia in Pigs Is Mediated by the Gastric-Hypothalamic-Pituitary Axis

The aim of the study was to determine the effects of gastric impairment in pigs on the axial and peripheral skeletal system properties and to test the hypothesis that fundectomy-evoked osteopenia is related to disturbed gastric-hypothalamic-pituitary axis function. Forty-day-old male piglets were subjected to experimental fundectomy (FX group, n = 6) to induce osteopenia, while sham operation was performed in the controls (SHO group, n = 6). At the age of 8 months, serum samples were collected, and the animals were sacrificed to obtain lumbar vertebrae (L1–L6) and right humerus for analysis. Using quantitative computed tomography (QCT) and dual-energy x-ray absorptiometry (DEXA) methods, bone mineral density and bone mineral content of the vertebrae and humerus were measured. The compression and three-point bending tests were applied to determine mechanical properties of lumbar vertebrae and humerus, respectively. Furthermore, geometric properties of humerus were assessed. Serum concentrations of ghrelin, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and selected macro- and microelements were also determined. Performed fundectomy decreased body weight in pigs by 66% compared with pair-fed sham operated pigs (P < 0.0001). Bone weight, bone mineral density, and bone mineral content of the lumbar vertebrae and humerus were significantly decreased in the fundectomized pigs (P < 0.01). Mechanical parameters of the lumbar spine and humerus were decreased after the fundectomy, as well. Serum concentrations of ghrelin, GH, and IGF-1 were lowered by 74.4%, 90.6%, and 54.6% in the fundectomized pigs, respectively (all P < 0.001). Moreover, the serum concentrations of calcium, magnesium, iron and copper in the fundectomized animals were significantly decreased by 15.5%, 45.3%, 26.7%, and 26.2%, respectively (P ≤ 0.05). In conclusion, the results obtained showed that both the disturbed gastric-hypothalamic-pituitary axis function and impaired mineral metabolism are associated with development of postfundectomy osteopenia of axial and peripheral skeleton in pigs.

Can Antibodies Against Glial Derived Antigens be Early Biomarkers of Hippocampal Demyelination and Memory Loss in Alzheimer’s Disease?

BACKGROUND Alzheimers disease (AD) is known to exhibit well characterized pathologies including the extracellular accumulation of amyloid plaques, intra-axonal presence of neurofibrillary tangles, and glial hypertrophy. Nevertheless, the nature of myelin pathology in AD has not been well studied. Recent studies on animal models of AD, however, revealed focal demyelination within amyloid-β plaques in hippocampus. OBJECTIVES In a view of this finding, we decided to assess humoral response against proteins of myelin sheath in AD, in the hope of identifying early biomarkers of memory loss and neuropathological process characteristic of AD. METHODS We assessed antibodies levels against proteins of the myelin sheath: myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP), myelin-associated glycoprotein (MAG), and proteolipoprotein (PLP) in sera of 26 AD patients and 26 healthy controls, using commercially available ELISA system (Mediagnost, Germany). RESULTS In the AD patient subgroup, significantly higher titers were observed for all types of assessed IgG autoantibodies compared to healthy control subjects (anti-MOG, anti-MAG, anti-MBP, anti-PLP). The titers of most of the investigated IgM antibodies were also higher in AD patients (p <  0.05), with the exception of anti-MAG IgM antibodies (p >  0.05). CONCLUSION The study provides the evidence for the significantly increased production of autoantibodies against proteins of myelin sheath in AD. These results can be of importance in the light of emerging data from animal models of AD, indicating early demyelination of hippocampal region. Further studies on larger population are necessary to confirm whether these autoantibodies could serve as early biomarkers of AD in humans.

Interrelationships between morphometric, densitometric, and mechanical properties of mandible in 6-month-old male pigs

Abstract The aim of the study was to evaluate interrelationships between final body weight, and morphometric, densitometric, and mechanical properties of the mandible in 6-month-old Polish Large White pigs exposed to dexamethasone and nanocalcium. The study was performed on 27 males, castrated on the 28th d after weaning, and reared until the age of 6 months, after which the animals were slaughtered, and the mandible was obtained. The pigs were divided into four experimental groups: animals given per os nanopartical calcium, animals injected with dexamethasone, animals given both nanopartical calcium per os and dexamethasone injections, and animals injected with placebo. After the slaughter, morphological properties of the mandible such as bone weight and length were determined. Using computed tomography technique, volumetric bone mineral density (vBMD) of the cortical bone (Cd), mean volumetric bone mineral density (MvBMD), and total bone volume (Bvol) of whole mandible were measured. Areal bone mineral density (BMD) and bone mineral content were evaluated with the use of dual-energy X-ray absorptiometric method. Using three-point bending test, mechanical parameters such as maximum elastic strength (Wy) and ultimate strength (Wf) of mandible were estimated. Pearson’s correlation coefficient (r) was determined between all the investigated variables. The obtained results showed a significantly positive correlation between body weight and mandible weight, mandible length, Bvol, Cd, BMD, BMC, Wy, and Wf. However, statistically insignificant correlations of MvBMD and body weight, mandible weight, mandible length, and Bvol were observed. Furthermore, Bvol and Cd were not found to be significantly correlated. In conclusion, this study showed numerous positive correlations between final body weight and densitometric, morphometric, and mechanical properties of the mandible. This bone of pigs may be used as an attractive model for further investigation on metabolic response of the skeleton to physiological, nutritional, toxicological, and pharmacological factors influencing bone tissue metabolism.

Diagnostic difficulties resulting from morphological image variation in spondylodiscitis MR imaging.

Summary Background: Spinal infection (discitis; spondylodiscitis) presents a wide spectrum of pathologies. The method of choice for spondylodiscitis imaging is magnetic resonance (MR). It provides detailed anatomical information, especially concerning epidural space and spinal cord. The main aim of this article is the description and evaluation of spondylodiscitis morphological variation visible in magnetic resonance imaging. Material/Methods: In this article we retrospectively analysed the patients diagnosed at the Department of Radiology of the Provincial Hospital No 2 in Rzeszów between October 2009 and October 2011. The subjects involved a group of five women aged 41–74 (mean 56.3 years) and eight men aged 46–69 (mean 61,3 years). All patients had spondylodiscitis symptoms. All patients underwent MRI examination before and after the contrast enhancement. In three patients additional CT examination was performed. Results: Following the MRI procedure all patients were diagnosed with typical symptoms of spondylodiscitis. It also revealed a number of pathologies resulting from morphological spondylodiscitis variation. Other pathologies found on the MR images of the study group patients involved epidural intra-canal spinal pathological masses causing spinal cord compression, lung abscess, pyothorax, paravertebral abscesses and epidural empyemas, abscess between adjacent vertebral bodies, abscesses beneath anterior longitudinal ligament, and iliopsoas muscle abscesses. In all cases a destruction of vertebral bodies with end plates loss restriction and cortical layer discontinuity was observed. Moreover, one person was diagnosed with pathological vertebral body fractures and liquefactive necrosis of the vertebral body. Conclusions: Spondylodiscitis manifests itself in a great number of morphological variations visible on the radiological images. Apart from ordinary features of vertebral bodies and discs, progressive spinal destruction is observed together with reactive bone changes and soft tissue infiltration. The latter leads to a number of complications e.g. abscesses or even fistulas and also to the formation of obstacles in radiological images. The knowledge of radiological images together with overall evaluation of clinical and laboratory features enables a proper diagnosis.