Activity of Lysosomal Enzymes During Protein Malnutrition and Progesterone Supplementation in the Mouse.

This study investigated the effects of protein malnutrition and progesterone supplementation on the activities of a spectrum of lysosomal enzymes in tissue fragments of mouse liver and kidney. The working hypothesis was that the known anti-stress action of progesterone could have to do with the inhibition of lysosomes which are engaged in apoptotic and oxidative stress-induced responses. The study investigated the effects of exogenous progesterone in chronically (3 weeks) protein-malnourished (10% protein) mice on the activities of lysosomal hydrolases in liver and kidney tissues. Progesterone was injected intraperitoneally in a dose of 2 µg/g body mass dissolved in a vehicle volume of 10 µL/g body mass during the final 3 days of exposure to either low 10% or standard 16% protein content in the chow. After euthanizing the animals, tissue fragments of liver and kidney assayed for the content of lysosomal enzymes. The results demonstrated the stimulating effect of protein malnutrition on lysosomal activities. We further found, contrary to our hypothesis, that progesterone supplementation during both standard and low-protein conditions enhanced lysosomal activities, particularly acting in concert with protein malnutrition in kidney tissue. The effects were selective concerning both lysosomal enzymes and tissues and of highly variable magnitude. Nonetheless, we believe we have shown that progesterone assists protein malnutrition in stimulation of lysosomal enzymes, which suggests the possibility of the hormone's engagement in cleansing the cellular milieu in disorders consisting of accumulation of toxic molecules.

Epidemiological Aspects of Low Back Pain.

Low back pain (LBP) is a major health problem, particularly in the contemporary societies of highly developed countries. This study seeks to define the influence of basic demographic and social factors, such as gender, body mass, physical activity, and the type of work, on the occurrence of lumbosacral spine pain in the early and middle-late adulthood. The study was based on a self-reported survey, using the revised Oswestry Low Back Pain Disability Questionnaire to evaluate pain symptoms, and managing everyday tasks. Physical activity was evaluated on the Minnesota Leisure Time Physical Activity Questionnaire. We found that patients in the early adulthood had a significantly lower level of disability. The older patients had a greater low back pain and motion, sleeping, and social life problems. Neither did gender nor the type of work, leisure time physical activity, or body mass appreciably affect the level of disability due to low back pain in both younger and older patient groups. We conclude that, all else unchanged from the epidemiological standpoint, wear and tear of the spine structure naturally progressing with age seems a major determinant of the appearance of low back pain.

The impact of organic vs. inorganic selenium on dairy goat productivity and expression of selected genes in milk somatic cells.

The aim of this study was to determine the effect of diet supplemented with selenized yeast (Se-yeast) on milk yield and milk composition of goats and expression of casein and mammary-gland-immune system genes in milk somatic cells (MSC). Twenty-four dairy goats in their second to fourth lactations were divided into control and experimental groups, balanced according to lactation number and breed (Polish White or Fawn Improved). Morning milk and blood samples were collected four times during lactation (on the 21st, 70th, 120th, 180th day after kidding). The control and experimental groups were fed diets with 0.7 mg inorganic Se/goat/day (sodium selenite) or 0.6 mg organic Se/goat/day (selenized yeast), respectively. Milk, fat and protein yields during lactation as well as average somatic cell count, fat, protein and lactose contents in milk were evaluated. Microelements in milk and blood serum and biochemical parameters in blood serum were determined at the beginning and the end of the experiment. The expression levels of the genes encoding αS1-casein (CSN1S1), αS2-casein (CSN1S2), κ-casein (CSN3), interleukin 8 (IL-8), serum amyloid A3 (SAA3), interleukin 1ß (IL-1ß), bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), ß2-defensin (GBD2), hepcidin (HAMP), chemokine 4 (CCL4), tumour necrosis factor α (TNFα), toll-like receptor 2 (TLR2), cathelicidin-7 (MAP34) and cathelicidin-6 (MAP28) were determined in MSC. Milk, fat, and protein yields were higher and somatic cell count (SCC expressed as natural logarithm) was lower in the milk of goats fed organic Se. The Se concentration in milk was twice as high in the organic vs. inorganic treatment groups at the end of the experiment, while there were no differences in studied biochemical parameters between groups. The transcript levels of CSN1S2 and BAC7.5 were higher and IL-8 was lower in MSC of Se-yeast treated groups. Such results may indicate better health status of mammary glands of goats treated with organic Se as well as positive impact of selenized yeast on the goat's milk composition. Differences in the IL-1ß and IL-8 transcript levels were also noted between the stages of lactation, with the highest expression at the peak of lactation (day 70), highlighting the metabolic burden at this time. We concluded that the Se-yeast supplementation improved the productivity and health status of goats and could have significant economic impact on farmer's income.

Effects of Glutathione on Hydrolytic Enzyme Activity in the Mouse Hepatocytes.

In this study, the effect of glutathione (GSH) on the activity of hydrolytic enzymes of lysosomal, microsomal, and cytosolic fractions was studied in the mouse hepatocytes. The experiments involved 30 Swiss male mice, divided into the experimental and control groups of 15 mice each. The former group received injections of 12 µL/g of GSH solution at a dose of 100 µg/g body weight, whereas the latter received 12 µL/g of physiological saline, all given intraperitoneally daily for 7 days. Then, fragments of liver tissue were collected from the euthanized animals and processed to obtain lysosomal, microsomal, and cytosolic fractions of hepatocytes. The activity of the following enzymes was investigated in vitro: ß-glucuronidase, ß-galactosidase, ß-glucosidase, N-acetyl-hexosaminidase, lysosomal esterase and lipase, acid phosphatase, cathepsin D and L, leucine aminopeptidase, and alanine aminopeptidase. We found that GSH, administered in vivo in the mouse, in the main (73% cases), increased the in vitro activity of the majority enzymes abovementioned, although the effect was somehow variable, depending on the fraction of hepatocytes and the type of enzyme. The findings imply that GSH supplementation may intensify the rate of cellular hydrolytic degradation, i.e., the rate of disposal by the cell of unwanted materials.

The Timing of Rehabilitation Commencement After Reconstruction of the Anterior Cruciate Ligament.

One of the most common injuries of the knee joint is a rupture of the anterior cruciate ligament (ACL). Most authors believe that early rehabilitation of patients after ACL reconstruction promotes better treatment outcomes. Less is known about the influence of the time that passes from injury to surgical reconstruction. Therefore, the goal of this study was to assess the dependence of treatment outcomes of ACL on injury-to-reconstruction and reconstruction-to-rehabilitation time lags. The study included 30 patients of the mean age 34 ± 7 years with trauma-related rupture of ACL and its surgical reconstruction. The time range from ligament rupture to its reconstruction was 120-180 days and from reconstruction to rehabilitation was 1-120 days. Postsurgical rehabilitation outcomes were assessed with the Lysholm knee scale and the IKDC 2000 subjective knee evaluation form. The scales were applied before and after rehabilitation. We found distinct improvements in all physical symptoms in the damaged knee joint, regardless of the time elapsed from trauma to ACL reconstruction and from ACL reconstruction to rehabilitation. The beneficial outcomes of rehabilitation were significantly inversely associated with the time elapsing from reconstruction to rehabilitation commencement but failed to depend on the time from ACL rupture to reconstruction. We conclude that rehabilitation should start as early as possible after ACL reconstruction to optimize the beneficial outcomes in terms of functional physical recovery, whereas the injury-to-reconstruction delay is less meaningful to this end.

Robot-Assisted Body-Weight-Supported Treadmill Training in Gait Impairment in Multiple Sclerosis Patients: A Pilot Study.

This study deals with the use of a robot-assisted body-weight-supported treadmill training in multiple sclerosis (MS) patients with gait dysfunction. Twenty MS patients (10 men and 10 women) of the mean of 46.3 ± 8.5 years were assigned to a six-week-long training period with the use of robot-assisted treadmill training of increasing intensity of the Lokomat type. The outcome measure consisted of the difference in motion-dependent torque of lower extremity joint muscles after training compared with baseline before training. We found that the training uniformly and significantly augmented the torque of both extensors and flexors of the hip and knee joints. The muscle power in the lower limbs of SM patients was improved, leading to corrective changes of disordered walking movements, which enabled the patients to walk with less effort and less assistance of care givers. The torque augmentation could have its role in affecting the function of the lower extremity muscle groups during walking. The results of this pilot study suggest that the robot-assisted body-weight-supported treadmill training may be a potential adjunct measure in the rehabilitation paradigm of 'gait reeducation' in peripheral neuropathies.

Activities of Lysosomal Enzymes in Alloxan-Induced Diabetes in the Mouse.

The study investigated a panel of lysosomal enzymes in the liver and kidney tissues in alloxan-induced diabetes in the mouse. The mice were divided into six experimental groups receiving 10% alloxan at a dose of 50 and 75 mg/kg over a period of four, eight, and twelve days; each group was compared with controls receiving 0.9% NaCl. The findings were that diabetes induced by both doses of alloxan was accompanied by significant increases in the lysosomal activities of acid phosphatase and the glycosidases investigated: ß-glucuronidase, ß-galactosidase, ß-glucosidase, and N-acetyl-hexosaminidase. The lysosomal enzyme activity in both liver and kidney cells peaked 12 days after onset of diabetes for most enzymes, at the time when hyperglycemia and hyperinsulinemia already started abating after their peak at 8 days into the course of diabetes. The enzyme activity was in most cases higher with the higher dose of alloxan and thus higher level of glycemia. Lysosomal enzymes degrade glycoconjugates, the molecules that are present in the basement membrane of endothelial cells where they contribute to capillary wall stability. Thus, enhanced activity of these enzymes could presage the progression of diabetic microangiopathy, atherosclerosis, and the development of microvascular complications.

Effect of ghrelin on activities of some lysosomal hydrolases in rabbits.

OBJECTIVES: Observing the changes of activity of some lysosomal enzymes in blood serum of female rabbits subjected to injection of 10 microg of ghrelin/kg of body weight. METHODS: In the blood serum the activity of cathepsins D and L, alanine aminopeptidase, acid phosphatase, lysosomal lipase and lysosomal esterase was determined. RESULTS: As a result of ghrelin injection the activity of all the enzymes examined in blood serum increased markedly. CONCLUSION: Changes of lysosomal enzymes activities in the blood serum caused by the effects of ghrelin should be regarded as the response of the lysosomal system.

Further structural analysis of GnRH complexes with metal ions.

MALDI-TOF mass spectrometry, 1H NMR spectrometry, the continuous variation method and molecular modeling by MM3 calculation confirmed our earlier studies showing that gonadotropin-releasing hormone (GnRH) forms complex with copper(II) ion with the binding ratio 1:1. The copper(II) complex formed at physiological pH has a square planar configuration and GnRH complexes with nickel(II) and cobalt(II) ions are less stable than that of copper(II).

Insulin-induced changes of proteolytic activity of the lysosomal enzymes.

OBJECTIVES: Changes in the activity of alanine aminopeptidase, leucine aminopeptidase and cathepsins D and L in the liver and kidney of male and female of mice, injected with 0.4 IU/kg b.w. insulin for 4 and 8 days. METHODS: The homogenates of the liver and kidney were taken for examination. The activity of alanine aminopeptidase, leucine aminopeptidase and cathepsins D and L has been determined according to [1] method. RESULTS: The activity of alanine aminopeptidase, leucine aminopeptidase, cathepsins D and L in the liver and kidney of male and female of mice decreased in effect of insulin injections for 4 and 8 days. CONCLUSION: The changes of enzyme activities showed a stimulating effect of the insulin injection on the labilization of lysosomal membranes. The range of the reaction remained in a relationship with the kind of the organ, the type of enzyme, time over which insulin introduced operates in the organism, and with the sex.

Effect of hydrocortisone on the activity of some lysosomal enzymes in mice.

OBJECTIVES: Changes in the activity of cathepsin D and L, alanine aminopeptidase, leucine aminopeptidase, N-acetyl-beta-glucosaminidase, lysosomal arylesterase and lysosomal lipase in the liver and kidney of unselected and selected mice, subjected to 7.5 mg/kg b.w. of hydrocortisone injection for 4 and 8 days. METHODS: The homogenates of the liver and kidney were subjected to differentiated centrifuging and determination of studied enzymes. RESULTS: Injection of hydrocortisone caused an increase in the activity of all investigated lysosomal enzymes in the liver and kidney of mice. CONCLUSION: The reactions of selected mice were stronger in comparison with unselected ones. The highest increase in the activity investigated enzymes was observed after 8 days of hydrocortisone injection.