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Introduction: Sarcopenia is a term for the loss of skeletal muscle mass, quality and performance associated with normal aging that can lead to frailty in the elderly. Contributing to physical frailty are skeletal muscle atrophy, declining strength, increasing fatigability and susceptibility to injury. For healthy elderly persons, the decrease in muscle mass results from a loss in the total number of fibers per muscle as well as a decrease in the mean cross sectional area of the remaining fibers. The aim of this study was to determine age-related changes in three human muscles with different function and location in the body. Materials and methods: The cross sectional area and the percentage of fibers type I, type IIA and type IIX were studied in vastus lateralis, deltoid and external intercostal muscle. Muscle samples were obtained from 30 male subjects, aged 20–80 years. Fiber types were defined immunohistochemically, using monoclonal antibodies specific for each type of fibers. Results: All muscle fiber types showed the reduction in the fiber size in all three examined muscles. In all muscles the proportion of type I and type IIA was changed, but not in type IIX. With increasing age results showed the increase in proportion of type I, while proportion of type IIA fibers decreased, with vastus lateralis muscle being the most affected. Conclusions: These results suggest that age-related muscle atrophy is not a general phenomenon, and does not affect all muscles equally.

Objective: Diabetes (lat. Diabetes mellitus DM) is now one of the most common diseases worldwide, affecting 415 million people. Abnormal glutamate homeostasis may contribute to the pathogenesis of diabetes with its direct and indirect effects on the integrity of the β-cells of pancreatic islets of Langerhans. The increased concentration of glutamate is present in all tissues with DM type 1 and type 2 DM. Polyphenols are associated with antioxidant, antihypertensive, anti-inflammatory, hypoglycemic and hypocholesterolemic effects on the human body. This study is based on the experimental therapeutic effect of olive leaves polyphenols in DM type 1. Oleuropein is a majority polyphenol in olive leaf extract (lat. Olea europaea L.). Material and methods: Type 1 DM was chemically induced in male Wistar rats by intraperitoneal injection of streptozotocin. The purpose of this study was to determine the level of glutamate in the liver of diabetic rats and to compare it to the level of glutamate in the liver of diabetic rats post-treated with different concentrations of polyphenols olive leaf extract (512 mg/kg, 768 mg/kg and 1024 mg/kg). The concentration of glutamate was determined spectrophotometrically in 20% liver homogenate while polyacrylamide gel, sodium dodecyl sulphate (SDS-PAGE) was used to separate the proteins and to determine the difference in the expression of the same in the experimental groups. Results: The increased levels of glutamate were established in the liver of diabetic rats. In groups of animals with short diabetes (8 days), glutamate concentration was proved to be lower compared to a group of animals with long-term diabetes (30 days). Furthermore, the results showed a significantly lower concentration of glutamate in the liver of post-treated animals with olive leaf extract in a dose-dependent manner.

Objective: The iliopsoas of the rat is composed of the psoas major muscle and the iliacus muscle and is generally considered to be a fast muscle. However, previous studies of the fiber type composition of the muscle showed different results. Therefore, the aim of our study was to examine the fiber type composition of the rat iliopsoas muscle with regard to its segmental origin. Materials and methods: At four different levels of its origin, using monoclonal antibodies for myosin heavy chain identification, the percentage and muscle fiber cross sectional areas (CSA) of Type I, IIA, IIB and IIX muscle fibers were identified. Results: The rat iliopsoas muscle had a predominance of fast muscle fiber types, with the highest percentage and the largest CSA of the fastest IIB muscle fibers. Moreover, our results showed changes in percentages, as well as the CSA-s of muscle fiber types in cranio- caudal direction in the psoas major muscle. There was a decrease of percentages and the CSA-s of type IIB muscle fibers from the cranial towards the caudal part. The results were published in the Journal of Anatomy (J Anat. 2017 ; 230(4):542-548.). Conclusions: The rat iliopsoas muscle is composed of all four muscle fiber types. It is a fast muscle with a predominance of fast type IIB muscle fibers with the largest CSA-s. The composition of the rat psoas major muscles changes in a cranio-caudal direction thus pointing to the more postural role of the caudal part of the muscle.

Kortikosteroidi izazivaju pojavu steroidnog oblika šećerne bolesti mnogobrojnim mehanizmima. Kortikosteroidi povećavaju otpornost na inzulin u stanicama jetre i stanicama drugih tkiva kao što su masno i mišićno tkivo. Imaju štetno djelovanje na beta-stanice gušterače, smanjenja unosa glukoze u stanice i indukcije apoptoze. Čimbenici rizika za razvoj steroidne šećerne bolesti su doza i dugotrajnost primjene kortikosteroida, dob, obiteljska anamneza šećerne bolesti, debljina, rasa i prethodno postojanje šećerne bolesti. Pristup pacijentima sa steroidnom šećernom bolešću je dijeta, tjelovježba i samokontrola glukoze u plazmi. Ako glikemija natašte ili nakon obroka naraste preko 11,1 mmol/l preporučena je primjena inzulina., The mechanism of corticosteroids-induced diabetes mellitus is multifactorial. Corticosteroids induce hepatic and extrahepatic insulin resistance. Corticosteroids have direct harmful effects on insulin-secreting beta cells of the pancreas, decreasing in glucose transport into the beta cells and inducing apoptosis. The risk of corticosteroids-induced diabetes increases with the corticosteroids dosage, duration of therapy, age, family history of diabetes mellitus, obesity, ethnicity and high blood glucose concentrations before corticosteroids therapy. Treatment of patients with corticosteroids-induced hyperglycaemia is diet, exercise and self-monitoring of blood glucose level. Patients with persistent fasting and daytime blood glucose concentrations over 11.1 mmol/L, the treatment with insulin is recommended.

UVOD M. iliopsoas u svinje građen je od dva mišića, m. psoas major koji polazi sa ventralne površine zadnjeg rebra (dio bliži kralješku) i poprečnih nastavaka svih lumbalnih kralježaka te m. iliacus koji polazi s ilijačne kosti. O građi ovog mišića postoji dosta kontroverznih rezultata. Imunohistokemijom nisu dokazana vlakna tipa IIB, slično kao i kod čovjeka iako neki autori navode da se izoforme teškog lanca miozina mogu ispoljavati istovremeno i na vlaknima tipa IIX i na vlaknima tipa IIB te takva vlakna nazivaju hibridnim vlaknima. S jedne strane, svinja je važna u medicinskim istraživanjima zbog sličnosti sa čovjekom, a s druge strane, za razliku od čovjeka koji hoda uspravno, svinja je veliki četveronožac. Stoga je cilj našeg istraživanja bio utvrditi građu m. iliopsoas u svinje i povezati je sa funkcijom. Ispitivanje je bilo provedeno na domaćim svinjama, teškim 120 kg žive vage. Iz lijevog i desnog m. psoasa i m. iliacusa uzimani su uzorci veličine 1x1x1 cm. Tipizacija mišićnih vlakana tipa I, IIA i IIX učinjena je imunohistokemijskom metodom bojanja teških lanaca miozina. ReZULTATI Rezultati su pokazali da je m.iliopsoas po distribuciji mišićnih vlakana brzi mišić odnosno mišić sa dinamičkom funkcijom. M. iliacus građen je pretežno od brzih vlakana tipa IIX koja su zastupljena više od 50%. Vlakna tipa IIA i tipa I zastupljena su u manjem postotku. Sličnu građu pokazuje i m.psoas major u kojem također prevladavaju brza vlakna tipa IIX. Najveću površinu poprečnog presjeka u m. ilacus imaju vlakana tipa IIX. Površine mišićnih vlakana tipa IIA i tipa I su podjednake i značajno manje od vlakana tipa I. U m. psoas major također brza vlakna tipa IIX imaju najveću površinu, spora vlakna tipa I nešto su manja, a najmanju površinu imaju brza vlakna tipa IIA. ZAKLJUČAK Usporedbom m.psoas major i m.iliacus utvrđeno je da u oba mišića prevladavaju brza vlakna (vlakna tipa IIA i IIX) što govori u prilog njegove dinamičke funkcije. Površina sva tri tipa mišićnih vlakana značajno je veća u m.iliacusu u odnosu na m.psoas major s time da najveću površinu poprečnog presjeka imaju vlakna tipa IIX.

The Reg genes have been well documented to play an important role in various tissue regeneration processes, cell proliferation, differentiation and carcinogenesis. However, the regulation of Reg gene expression in rat skeletal muscles regeneration process is poorly understood. Therefore, the aim of this investigation was to reveal the transcriptional activation of Reg genes in the skeletal muscles in response to injury. In 2-month-old Wistar rats three experimental models of muscle and nerve injury were used. The regeneration process was induced in both types of muscles, slow (m. soleus, SOL) and fast (m. extensor digitorum longus muscle, EDL) by injection of 1 ml of local anesthetic, by denervation while in third experimental model muscle denervation and injection of local anesthetic were combined. The RT-PCR analysis was performed for detection of Reg1, Reg3G and Reg4 mRNA in regenerating muscles. RT-PCR analysis showed that Reg1 gene is expressed in EDL and SOL with peak expression 12 hours after injury and signal intensity was constant till 7th day in all three experimental models. Reg3G can be detected over a longer period in comparison with Reg1 and signal intensity was lower. There was no detectable Reg4 mRNA expression in either SOL or EDL muscle. Our investigation revealed presence of Reg1 and Reg3G genes in regenerating skeletal muscle and its expression is up regulated with regeneration process. Reg1 and Reg3G genes expression was induced in both skeletal muscles at a considerably early time point after muscle injury. Our results imply that Reg genes have a significant role at the early stage of skeletal muscle regeneration.

Increased oxidative stress and impaired antioxidant defence are important mechanisms in the pathogenesis of diabetic neuromyopathy. Since diabetes mellitus type 1 decreases muscle regeneration capacity the present study was designed to determine the influence of alpha-lipoic acid (LA), a potent biological antioxidant, on the process of regeneration of diabetic rat skeletal muscles. The regeneration process was provoked in streptozotocin induced diabetic rats in slow (m.soleus, SOL) skeletal muscles by intramuscular injection of myotoxin bupivacain. At intervals of ten days and four weeks, muscle histochemical and morphometrical analysis (fibre cross areas and fibre type distribution) was performed. Changes induced by diabetes are evident in redistribution of muscle fibers and in significant level of atrophy. After four weeks of diabetes, type II muscle fibers showed higher percentage in respect to control group. Slow muscle treated with LA showed a complete recovery with significant higher percentage of type I fibers. Muscle atrophy is present in all fiber types of slow skeletal muscle. Treatment with LA induced significant increase in cross-sectional area in both fiber types. Our results have shown that treatment with LA can reduce changes in the morphological properties caused by diabetes mellitus in slow rat skeletal muscles during the process of regeneration. Treatment with LA during four weeks has shown effects on the redistribution of muscle fibers, and can prevent atrophy in slow diabetic skeletal muscle.

The Reg genes play a major role in the regeneration of various tissues ; however, no reports have been published regarding expression of the Reg3G gene in skeletal muscle. In this study we investigated the expression of the Reg3G gene in regeneration of rat skeletal muscle and injured nerves. We used 3 experimental models of muscle and nerve injury. RT-PCR and Western blot analysis were performed for detection of Reg3G in regenerating muscle and nerve. We found transcriptional activation of the Reg3G gene in the soleus and extensor digitorum longus muscles and in their corresponding nerves after both muscle and nerve injury in different time periods, respectively. The results suggest that the Reg3G gene plays a major role in communication between injured axons and muscle and may play a significant role in skeletal muscle and peripheral nerve regeneration.

The aim of this study was to establish a 14-day time course study on innervated and denervated rat soleus and tibialis anterior muscles after treatment with salbutamol, a 2 adrenoceptor agonist to assess fiber type selectivity in their type composition. Soleus, as a slow-twitch muscle, is composed of 90% type I fibers and 10% type II fibers. On the other hand, tibialis anterior muscle is fast-twitch with 91, 6% type II fibers and 8, 4% type I fibers. We observed that during the period of denervation occurs the conversion from type I to type II muscle fibers. However, in soleus muscle of the animals which were treated with salbutamol this conversion was less expressed and an increased number of type I fibres were observed. In tibialis anterior muscle, fibres conversion was higher what caused the significant increase in number of type II fibres. Groups of healthy muscles that were treated with salbutamol, showed an increase in number in both muscles of fibres type II until 9th day of experiment. While in soleus the number of fibres type II was increasing towards the end of experiment, in tibialis anterior muscle showed a conversion in second part of experiment from type II fibers to type I. In this study, we have shown how muscles of different properties reacted differently to various stimuli.

Depression and anxiety are co-morbid condition in diabetes as disease-related psychological reactions on this chronic metabolic illness. This study was aimed to determine the occurrence of depression and anxiety in seafarer’s type 2 diabetic patients. A random sample of 52 diabetic seafarers treated with diet and oral glucose lowering agents, and 56 healthy seafarers were screened for depression with The Beck Depression Inventory (BDI) and for anxiety with State-Trait Anxiety Inventory (STAI 1, STAI 2). Depression (BDI > 18, 5) and anxiety (STAI < 28, 5) was significantly higher in the group of diabetic seafarers than in control group (more than 30%). Significant correlation was noted between depression and duration of diabetes mellitus, degree of obesity and poor glycaemic control (HbA1C>8%) and longer duration of shipping routes (over 6 months). The proportion of depression and anxiety was found higher in seafarer’s type 2 diabetic patients than in the healthy seafarers.

INTRODUCTION In 1988, Terazono et al. first isolated a gene expressed in regenerating pancreatic β cells which he named regenerating gene (Reg) (Terazono et al. 1988). Several reg and reg-related genes have been isolated from mammalian species and they constitute a multigene family, the Reg gene family. Based on the primary structure of the proteins encoded by Reg family genes, the members of the Reg family have been grouped into four subclasses, type I, II, III and IV (Okamoto 1999, Hartupee 2001). Reg1 was originally isolated from a rat regenerating islet-derived cDNA library (Terazono et al. 1988) and it acts as a growth factor for pancreatic β cells in an autocrine/paracrine manner (Okamoto 1999). Reg1 protein expression has also been found in the rat gastrointestinal tissues and kidney (Watanabe et al. 1993). Since there are no studies showing Reg1 expression in healthy or injured skeletal rat muscles, the aim of this investigation was to reveal the Reg1 gene expression in regenerating skeletal muscle. Previous studies reported that there was no expression of Reg1 gene after peripheral nerve axotomy. Therefore, in this study, we examined the expression of Reg1 gene of corresponding nerve in response to skeletal muscle injury. MATERIAL AND METHODS In two-month old Wistar rats regeneration process was induced in slow (musculus soleus, SOL) and fast (musculus extensor digitorum longus, EDL) type of muscle. Muscle regeneration was induced by injection of local anesthetic (0.5% bupivacain). Immunohistochemistry and PCR analysis where performed for detection of Reg 1 mRNA in untreated and regenerating SOL, EDL and in sciatic nerve after 6, 12, 24 hours, 3 and 7 days after regeneration process was induced. Immunohistochemical staining for anti-rat Reg 1 protein Mab. Tissue samples of rat muscles and nerves were fixed in cold acetone. The cryostat sections were incubated with an anti-rat Reg 1 protein antibody overnight at 4°C and with secondary antibody at room temperature for 30 min. Bound antibody was detected by universal immunoenzyme polymer method followed by hematoxylin staining. RT-PCR. Total RNA was extracted from samples using Promega Total RNA extraction kit (SV Total RNA Isolation system, Promega, Madison, WI, USA). 1000ng of total RNA from each sample was transcribed in cDNA using Superscript II RT (Invitrogen, Carlsbad, CA, USA). Reg1 primer sequences were 5'-TACAGCTGCCAATGTCTGGATT-3' and 5’-CAGTGTCCCAGGATTTGTAGAGA-3’. The amplification products were then separated by 3% agarose gel electrophoresis stained with Gel Red (Biotium, Hayward, CA, USA) and visualized under UV light. Results were quantified with densitometry method by «1D Image Analysis Software» (Eastman Kodak Company, Rochester, NY, USA). RESULTS Immunohistochemical analysis of SOL and EDL muscles after three days of regeneration process was induced showed Reg I positive cells. (Figure 1, 2) RT-PCR analysis showed that Reg1 gene is expressed in both types of skeletal muscles and sciatic nerve. EDL has a peek expression 12 hours after regeneration process was induced and the signal intensity was stronger but not statistically significant compeering to SOL. After 12 hours the signal intensity was rapidly decreased till the 3rd day. This time point has importance because of statistically significant difference in signal between SOL and EDL. Signal is weak but constant till 7th day. SOL has also peek expression 12 hours after injury, although the signal is more constant and is weakening until the day 7th. Sciatic nerve for instance has no detectable expression 6 hours after injury and first measurable signal has been detected after 12 hours with peek expression 24 hours after the local anesthetic has been applied. After 24 hours the signal is weakening and could not be detected 7 days after regeneration process was induced.

AIM The family of muscle specific basic helix-loop-helix (bHLH) regulatory factors consists of MyoD, Myf-5, myogenin and MRF4. These factors contribute to the transcriptional activation of muscle genes during commitment and differentiation. A distinct role for the bHLH myogenic factors initially was suggested by their different temporal expression patterns in the somites and limbs during embryonic development. The aim of this study was to determinate time course expression of MyoD and myogenin during regeneration in muscles of different developmental origin and different rat strains. MATERIALS AND METHODS All experiments were performed on male Wistar rats and male Sprague-Dawley rats. Histochemical, immunohistochemical and Western blot analysis were performed on m. tibialis anterior and m. masseter. Regeneration process of skeletal muscles was induced by injection of bupivacain. RESULTS AND CONCLUSIONS We have demonstrated that muscle regeneration caused a transient upregulation of MyoD and myogenin, but after reestablishment of normal muscle architecture there was no expression of MyoD and myogenin. Time course expression of MyoD and myogenin during regeneration in muscles of different developmental origin was similar. The main conclusion of our investigation is that the role of MyoD and myogenin is potentially as important in managing an adaptive and regenerative response of muscle fibers.

Introduction Proportion of muscle fibres in the muscle depends on its primary function. Psoas muscle has complex dynamic as well as postural function. It connects lumbar spine with lower limb and therefore acts in both, the spine and the leg. The aim of presented study was to determine the distribution of different fiber types in the human psoas muscle by using immunohistochemistry and explore muscles composition relating to its complex function. In addition, we wanted to compare the composition of the muscle with respect to the left and right side, as to determine difference according to dominant side of the body. Methods Muscle samples were collected from 30 adult males. Serial transverse sections (5µm) of the samples were cut by cryomicrotome. Type I, IIA and IIX muscle fibres were typed using myosin heavy chain (MHC) identification. Serial sections were analyzed using light microscope with magnitude of 100x. Results Our study showed predominance of type IIA muscle fibers (48.34%) in the human psoas major muscle, type I fibres were present at 40.72% and type IIX fibres were present at 10.94%. The biggest cross-sectional area (CSA) had type I fibres (2571.12µm2), type IIA fibres had a CSA of 2025.44µm2 and type IIX fibres had the smallest CSA (1706.77µm2). There were no differences between composition of the left and right psoas muscle. Conclusion We concluded that fibre type composition of the psoas muscle indicates its dynamic and postural functions which supports the fact that it is main flexor of hip joint (dynamic function) and stabilizer of lumbar spine, sacroiliac and hip joint (postural function).

Muscle fibers are dynamic structures capable of altering their phenotype under various pathological conditions. The aim of the present study was to investigate the influence of long-lasting diabetes mellitus on the process of muscle regeneration in the skeletal muscle. Wistar rats were made diabetic by a single intraperitoneal injection of streptozotocin (STZ). The regeneration process in the skeletal muscle was induced in slow (m. soleus, SOL) and fast (m. extensor digitorum longus, EDL) muscles by injection of local anesthetic (bupivacaine). Skeletal muscles were analyzed 10 days, 4 and 8 weeks after bupivacaine treatment. Diabetes mellitus has changed morphological properties of both slow and fast skeletal muscles during the process of regeneration. These changes are evident in redistribution of muscle fibers and significant level of atrophy. All fiber types of diabetic fast muscles showed stronger atrophy than muscle fibers in slow muscles which have more oxidative metabolism. The changes of redistribution of muscle fibers depend on duration of diabetes and affect all types of muscle fibers.

Sažetak Cilj: Cilj je ovoga rada bio utvrditi koji mišići sudjeluju pri izvođenju pojedinih pilates vježbi. Uz to smo željeli ustanoviti ima li pilates utjecaja na posturu, disanje i kondiciju te na poboljšanje medicinskih problema vezanih uz lokomotorni sustav. Metode: Istraživanje je provedeno na skupini od 30 ispitanica u dobi od 20 do 50 godina, koje vježbaju isključivo pilates metodom dva do tri puta u tjednu. Za vježbanje pilatesa koristili smo švicarsku loptu te utege od pola kilograma. Setove vježbi podijelili smo u tri skupine: vježbe za gornje ekstremitete, vježbe za trup i leđa te vježbe za donje ekstremitete. Rezultati: 41% ispitanica ima između 30 i 40 godina, 43% ispitanica ima visoku stručnu spremu, 63% ispitanica odlučilo se za ovu metodu vježbanja zbog rekreacije i održavanja tjelesne kondicije, a njih 37% zbog medicinskog problema. O djelovanju pilatesa na poboljšanje njihova zdravstvenog stanja, čak 83% ispitanica odgovorilo je pozitivno, 77% ispitanica izjasnilo se da vježbe pozitivno djeluju na njihovo raspoloženje i koncentraciju, čak 90% ispitanih žena misli da su vježbe poboljšale njihovo držanje tijela. Zaključak: Svakom pilates vježbom aktivira se velik broj mišića, razvija se snaga i fleksibilnost i učvršćuju se mišići odgovorni za potporu koštano-zglobnih struktura., Summary Aim: The aim of our study was to determine which muscles are active during the performance of the Pilates exercise. Beside that, we were investigated whether Pilates exercise influence on body posture, respiration and physical condition. Also we were studied the effect of this exercises on different pathophysiological condition of the musculoskeletal system Methods: The study was performed on 30 females aged 20-50 years old who practiced Pilates two to tree times per week. For Pilates performance they used swish ball and weights of a half of a kilogram. The sets of exercises were divided into three groups: exercises for the upper extremities, exercises for the trunk and back and exercises for the lower extremities. Results: The results of the survey show that 33% of the subjects were aged 20-30 years, data show that 43% of subjects had university education, 63% of the subjects took up the exercise method in order to maintain physical fitness and recreation, 37% due to some medical conditions. The answers to the question about the effects of exercises on the improvement of their health were positive in as much as 83%, 77% of the subjects believed that the exercises had a positive influence on their mood and concentration, as much as 90% of the surveyed women thought that exercises improved their posture. Conclusion: Each Pilates exercise activate great number of muscles, straight the muscle power and flexibility and as a results the muscles responsible for supporting skeletal structure were stronger.

Objective: The Reg I gene is expressed during regeneration of various tissues but there are no studies showing its expression in skeletal muscle. The aim of our study was to explore the expression of Reg I gene in healthy skeletal muscles and to define the time course of Reg I gene up- and down-regulation during muscle regeneration after muscle injury in the rat slow and fast skeletal muscle. Material and methods: In 2-month-old Wistar rats, the regeneration process was induced in slow (musculus soleus, SOL) and fast (musculus extensor digitorum longus, EDL) types of skeletal muscle. Muscle degeneration/regeneration was induced by injection of local anesthetic (0.5% bupivacain). The immunohistochemistry and western blot analysis were performed for detection and quantification of Reg I protein in regenerating skeletal muscles after 1, 2, 3, 4, 5, 6, 7, 10 and 15 days of regeneration. Results: Reg I gene was not expressed in adult, healthy SOL and EDL muscles while its expression was induced during the process of muscle regeneration (in both SOL and EDL). The peak of Reg I expression was on third day following the start of the regeneration process. The Reg I gene was down-regulated by day 5 and disappears by day 6-7. Conclusions: The Reg I gene is not expressed in normal rat skeletal muscle. The rapid switching in Reg I gene expression takes place during the early stages of muscle regeneration in the rat SOL and EDL muscles.

Background: Microdialysis continuous glucose monitor (GlucoDay) provides assessment of blood glucose dynamics. Patients and Methods: In 52 type 1 diabetics on two times/day NPH and pre-meal aspart insulin randomized in two groups: the continuation of NPH (n = 26), or once daily glargine (n = 26), 48-h GlucoDay registrations were started on the beginnig and after 4 months of the following. Results: At baseline, time spent in the euglycemic range (glucose between 3.9 and 8.0 mmol/l) was for the NPH group 37.96 ± ; 6.81% and 35.83 ± ; 6.24% for the glargine group. At end point, time in the euglycemic range increased in both groups (51.02 ± ; 7.22% versus 57.29 ± ; 10.27%). Time spent in the hypoglycemic range (glucose below 3.9 mmol/l) was 9.98 ± ; 2.57% for the first group, and 10.24 ± ; 3.55% for the second group at baseline. At end point, time in the hypoglycemic range decreased in both groups (8.00 ± ; 2.13% versus 6.59 ± ; 2.04%). Conclusion: The analysis of the GlucoDay data gave us other informations about glycemia than hemoglobin A1c and self-monitoring of blood glucose, such us peakless activity profile and the lower percentage time spent in the hypoglycemic range in the glargine treated group.

The organic hyperinsulinism was detected in a 52 year-old man but with no localization of pancreatic functional endocrine tumor using the standard noninvasive imaging studies (ultrasonography, computed tomography). An insulinoma is a rare tumor, typically less than 2 cm in diameter. The period of 15 years with symptoms of hypoglycaemia was passed before the tumor was evident with imaging studies (endoscopic ultrasound and abdominal computed tomography). The operative and pathologic findings confirm the diagnosis of insulinoma as well-differentiated functional endocrine tumor with benign biologic behaviour.

Background and Aims: Morphological changes of diabetic skeletal muscles are induced by diabetic neuropathy. The important role in the progression diabetic neuromyopathy have increased oxidative stress and impaired antioxidant defense. It is well known that alpha-lipoic acid (LA), as potent biological antioxidant, used in therapy of diabetic neuropathy. The aim of our study was to determine the influence of alpha-lipoic acid on the diabetic skeletal muscle, using the model of muscle rengeneration. Material and methods Diabetes were induced in two-months-old male Wistar rats by a single intraperitoneal injection of streptozotocin (STZ). Regeneration process was induced in slow (m.soleus, SOL) and fast (m.extensor digitorum longus, EDL) muscles by injection of local anesthetic (bupivacain). All groups of rats (control, untreated diabetic and treated diabetic) were treated with LA intraperitoneal injection every day (50 mg/kg). Skeletal muscles were analysed (fibre cross areas and fibre type distribution) 10 days, 4 and 8 weeks after bupivacaine treatment. Results: M. Soleus After 4 weeks of diabetes the percentage of type I fibers in SOL muscles was significantly lower in respect to control group, while the percentage of type I and IIA of muscle fibers in the diabetic group treated with LA was similar to these in control group. The fibre cross areas of type IIA diabetic muscle fibers are significant smaller in respect to normal muscles. After 4 weeks of treatment, the fibre cross areas of type I and type IIA diabetic muscle fibers were significantly higher. M. Extensor Digitorum Longus Diabetic untreated muscles, 4 weeks after regeneration, showed statisticaly increased percentage of type IIX fibers. In diabetic EDL muscles treated with LA were statisticaly more fibers type I and IIB. The cross areas of all fibre types in diabetic EDL muscles were significantly smaller than in control muscles. In diabetic group treated with LA the fibre cross areas were significantly higher than in diabetic group. Conclusions: The antioxidant therapy can prevent the canges in fiber type distribution and cross sectional areas of diabetic regenerating slow and fast skeletal muscles.

Background and Aims: Muscle fibers are dynamic structures capable of altering their phenotype under various physiological and pathological conditions. Skeletal muscle is composed of different fiber types (type I, IIA, IIX and IIB) with unique metabolic and contractile characteristics. Diabetes mellitus changes histochemical and morphometrical characteristics of skeletal muscle because of the influence on many neurogenic and myogenic factors. The progression of diabetes mellitus is concomitant with decrease of the regenerative capacity of skeletal muscle and treatment with insulin can prevent further atrophy of slow and fast skeletal muscles. This study was intended to explore the effects of type 1 diabetes and its treatment with insulin plus antioxidant (alpha-lipoic acid - LA) on the the process of skeletal muscle regeneration because oxidative stress have been implicated in pathogenesis of diabetic myopathy. Material and methods: Diabetes was induced by a single streptozotocin (STZ) 65 mg/kg with i.p. injection. Two-months-old male Wistar rats were randomized to control group, untreated diabetes group, diabetes group treated with insulin and diabetes group treated with insulin plus LA (50 mg/kg once daily). Regeneration process was induced in slow (m.soleus, SOL) and fast (m.extensor digitorum longus, EDL) muscles by injection of local anesthetic (bupivacain). The morphometric analysis (fibre cross areas and fibre type distribution) of skeletal muscles was performed after 10 days and 4 weeks. Results: Soleus muscle. In control group fiber, after 4 weeks of regeneration type distribution was similar to normal SOL without the regeneration. In the same period diabetic muscles, the percentage and the fibre cross areas of type I and IIA, did not reach values of control and normal SOL without the process of regeneration. In the group treated with insulin and insulin plus LA, the morphometric analysis of muscle was similar to these in control group. Extensor digitorum longus After 4 weeks of experiment control muscles were completely regenerated. In diabetic untreated muscles after 4 weeks of diabetes the percentage of type I and IIA fibers was greater. The treatment with insulin and insulin plus LA prevented changes in fiber type distribution during all investigated periods. Insulin therapy, in all experimental periods, resulted in a significant increase in fibre cross areas all fibre types. Additional therapy with LA leaded to better correction of the cross areas all fibre types like in control group and normal SOL without the process of regeneration. Conclusion: The additional use of antioxidant therapy with insulin has the potential benefit in the treatment of diabetic myopathy.

Background: Recent studies had shown that early phases of diabetes development is associated with oxidative stress and atherosclerosis. In type 2 diabetes atherosclerosis development have been related with high ferritin and low folate levels. The aim of this study is to investigate contribution of these two parameters on development of atherosclerosis in early phases of diabetes (pre-diabetes). Patients and Methods: This study included over random sample of 52 patients: 17 male (34%) i 34 females (66%). They were separated in two groups: group N – patients with normoglycaemia and group D - patients with prediabetes (45%). Results: 52 patients were examined ; mean age of group N=63, 42&plusmn ; 10, 9 years ; group D=64, 54&plusmn ; 8, 9 years. Values of HBA1C were in group N=4, 67%&plusmn ; 0, 3 ; in group D=4, 96%&plusmn ; 0, 4 ; ferritin in group N=74, 1μ g/mL&plusmn ; 18, 2 ; in group D=134, 1μ g/mL&plusmn ; 16, 2. The levels of folate were in group N=10, 9μ g/mL&plusmn ; 6, 2 ; in group D=8, 1μ g/mL&plusmn ; 3, 2. The results indicated statisticaly higher index ferritin / folat (p

Background and Aims: The progression of diabetes mellitus is concomitant with decrease of the regenerative capacity of skeletal muscle and treatment with insulin can prevent futher atrophy of slow and fast skeletal muscles. The aim of this study was to explore the effectss of type 1 diabetes and its treatment with insulin plus antioxidant (alpha-lipoic acid - LA) on the the process of skeletal muscle regeneration because oxidative stress have been implicated in pathogenesis of diabetic myopathy. Material and methods: Diabetes was induced by streptozotocin. Wistar rats were divided in four groups: control, diabetes, diabetes treated with insulin and diabetes treated with insulin plus LA. Regeneration process was induced by injection of local anesthetic. Analysis od fibre cross areas and fibre type distribution was performed after 10 days and 4 weeks. Results: Soleus muscle. After 4 weeks of regeneration , fiber type distribution of control group was similar to normal soleus without the regeneration. After 4 weeks of regeneration , the percentage and the fibre cross areas of type I and IIA of diabetic muscles, did not reach values of control and normal soleus without the process of regeneration. In the group treated with insulin and insulin plus LA, the morphometric analysis of muscle was similar to these in control group. Extensor digitorum longus muscle. After 4 weeks of experiment control muscles were completely regenerated. In diabetic untreated muscles after 4 weeks of diabetes the percentage of type I and IIA fibers was greater. The treatment with insulin and insulin plus LA prevented changes in fiber type distribution during all investigated periods. Insulin therapy, in all experimental periods, resulted in a significant increase in fibre cross areas all fibre types. Additional therapy with LA restored the cross areas all fibre types. Conclusion: The additional use of antioxidant therapy with insulin have the potential benefit in the treatment of diabetic myopathy.

Skeletal muscle is an extremely heterogeneous tissue composed of four fibre types with different sensitivity to insulin action. Diabetes mellitus type 1 is associated with neuromyopathy. Increased oxidative stress and impaired antioxidant defence are important mechanisms in the pathogenesis of this diabetic complication. Regeneration capacity of skeletal muscle during diabetes mellitus is reduced. Since alpha-lipoic acid (LA) is potent biological antioxidant the aim of present study was to determine the influence of LA on the process of muscle regeneration. Two-months-old male Wistar rats were randomised to control group, untreated diabetes group and diabetes group treated with LA. Diabetes was induced by i.p. injection of streptozotocin (STZ). Regeneration process was induced in slow (m.soleus, SOL) and fast (m.extensor digitorum longus, EDL) skeletal muscles by injection of local anaesthetic, bupivacain. At intervals of 10 days and 4 weeks, morphometric analysis of skeletal muscles was performed. The percentage of type I fibres in diabetic SOL muscles was significantly lower in respect to control group after 4 weeks of diabetes. In the diabetic group treated with LA the percentage of type I and IIA are similar to those in control group. In diabetic soleus muscle treated with LA, the fibre cross areas of type I and type IIA were significantly higher after 4 weeks of treatment in respect to diabetic muscles. Diabetic untreated EDL muscles, 4 weeks after regeneration, showed statistically increased percentage of type IIX fibres. In diabetic EDL muscle treated with LA there was statistically more fibres of type I and IIB. In diabetic EDL muscle, the cross areas of all fibre types were significantly smaller than in control muscles. In diabetic group treated with LA the fibre cross areas were significantly higher than in diabetic group. Our results suggest that treatment with LA can restore fibre type distribution and cross sectional areas induced by diabetes mellitus in regenerating slow and fast skeletal muscles.

Background and Aims: Morphological changes of diabetic skeletal muscles are induced by diabetic neuropathy. Increased oxidative stress and impaired antioxidant defense are important machanisms in the pathogenesis of diabetic neuromyopathy. Since it has been demonstrated that alpha-lipoic acid (LA) is a potent biological antioxidant the aim of our study was to determine the influence of alpha-lipoic acid on the the process of muscle regeneration. Material and methods:Two-months-old male Wistar rats were made diabetic by a single intraperitoneal injection of streptozotocin (STZ). Regeneration process was induced in slow (m.soleus, SOL) and fast (m.extensor digitorum longus, EDL) muscles by injection of local anesthetic (bupivacain). All groups of rats (control, untreated diabetic and treated diabetic) were treated with LA (50 mg/kg). Skeletal muscles were analysed (fibre cross areas and fibre type distribution) 10 days, 4 and 8 weeks after bupivacaine treatment. Results: M. Soleus - After 4 weeks of diabetes the percentage of type I fibers in SOL muscles was significantly lower in respect to control group, while the percentage of type I and IIA of muscle fibers in the diabetic group treated with LA was similar to these in control group. The fibre cross areas of type IIA diabetic muscle fibers are significant smaller in respect to normal muscles. After 4 weeks of treatment, the fibre cross areas of type I and type IIA diabetic muscle fibers were significantly higher. M. Extensor Digitorum Longus - Diabetic untreated muscles, 4 weeks after regeneration, showed statisticaly increased percentage of type IIX fibers. In diabetic EDL muscles treated with LA were statisticaly more fibers type I and IIB. The cross areas of all fibre types in diabetic EDL muscles were significantly smaller than in control muscles. In diabetic group treated with LA the fibre cross areas were significantly higher than in diabetic group. Conclusions: These results suggest that treatment with LA can restore fiber type distribution and cross sectional areas induced by diabetes mellitus in regenerating slow and fast skeletal muscles.

Background and aims. Insulin is a major anabolic hormone in the body, and its deficiency causes diabetes mellitus. It is known that skeletal muscle requires insulin for normal growth and development. Adult skeletal muscle is composed of muscle fibres that differ in their functional and molecular properties. This diversity is based on the pattern of expression of specific myosin heavy chain (MHC) isoforms. Using monoclonal antibodies specific to MHC isoforms, we can distinct two major fibers types: slow contracting fibers, called type I, containing slow MHCs, and three types of fast contracting fibers, called IIA, IIX and IIB muscle fibers. The aim of present study was to investigate the effect of diabetes mellitus type I on skeletal muscle during the four-week period. Material and methods. Two-month old male Wistar rats were randomized to control group and diabetes group. Diabetes was induced by a single streptozotocin (STZ) 65 mg/kg with i.p. injection. At interval of 4 weeks, morphometric analysis (fibre cross areas and fibre type distribution) of skeletal muscles was performed. Results. In diabetic muscles, comparing to normal muscles, the fibre cross areas of all four fibre types (I, IIA, IIX and IIB) were significantly smaller than in normal muscles (P

Background and Aims: Skeletal muscle is composed of different fiber types (type I, IIA, IIX and IIB) with unique metabolic and contractile characteristics. Muscle fibers are dynamic structures capable of altering their phenotype under various physiological and pathological conditions such as: altered hormonal profiles, increased or decreased neuromuscular activity, aging, etc. It is well known that skeletal muscle has the remarkable ability to complete a rapid and extensive regeneration process in response to pathological stimulus. The aim of the present time-course study was to investigate the influence of long-lasting diabetes mellitus type 1 on the process of muscle regeneration in fast and slow rat skeletal muscle. Material and Methods: Two-months-old male Wistar rats were randomly assigned to control and diabetic group. Rats were made diabetic by a single intraperitoneal injection of streptozotocin (STZ). Regeneration process was induced in slow (m.soleus, SOL) and fast (m.extensor digitorum longus, EDL) skeletal muscles by injection of local anaesthetic (bupivacain) in both groups. Fiber type distribution and fiber type areas were analysed after 10 days, 4 and 8 weeks of bupivacaine treatment in both SOL and EDL muscles. Results Changes in fiber type distribution Ten days after bupivacaine treatment control SOL muscle is composed of 89% of type I fibers and 11% of type II A fibers. In contrast, the percentage of type I fibers in diabetic SOL muscles was lower in respect to control group after 4 weeks of diabetes, while type I fibers became predominant after 8 weeks of regeneration. Ten days after bupivacaine treatment control EDL muscle is composed of 9% of type I fibers, 24% of type II A, 10% of type II X, and of 57% of type II B fibers. After 8 weeks of regeneration diabetic EDL muscles were composed of fibers with mainly oxidative metabolism i.e. type I and type IIA fibers. Changes in fiber cross areas of diabetic muscles Diabetic muscles, both SOL and EDL, showed a considerable degree of muscle atrophy at all time-point of muscle regeneration comparing to normal muscles. After 8 weeks of diabetes type I fibers showed statistically significant reduction in cross-sectional areas, while type IIA fibers completely disappeared. In diabetic EDL muscles the cross areas of all fiber types were significant smaller than in normal muscles in all investigated periods of regeneration with exception of type I fibers after 10 days of diabetes. Conclusions We have found that diabetes mellitus type 1 have changed the morphological properties in slow (SOL) and fast (EDL) muscles during the process of regeneration. These changes are evident in redistribution of muscle fibers and significant level of muscle atrophy. In diabetic EDL muscles all fiber types showed stronger atrophy than muscle fibers in slow muscles which have more oxidative metabolism. The changes of redistribution of muscle fibers depend on duration of diabetes.

Diabetic neuromyopathy is charactized with muscle weakness and muscle wasting. Skeletal muscle requires insulin for normal growth and development. It is composed of district fiber types with unique metabolic characteristics and heterogenity of insulin action (slow-contracting fibers, called type I, and three different types of fast-contracting fibers, called IIA, IIX and IIB muscle fibers). It has been shown that progression of diabetes mellitus is concomitant with decrease of the regenerative capacity of skeletal muscle. The aim of present study was to investigate the influence of insulin during the long term diabetes on the process of slow and fast skeletal muscle regeneration. Two-months-old male Wistar rats were randomised to control group, untreated diabetic group and diabetic group treated with insulin. We used a single streptozotocin (STZ) 65 mg/kg with i.p. injection for inducing experimental diabetes. Muscle regeneration was induced by injection of local anesthetic, bupivacain in slow (m.soleus, SOL) and fast (m.extensor digitorum longus, EDL) skeletal muscles. Morphometric analysis (fibre cross areas and fibre type distribution) of skeletal muscles was performed at intervals of 10 days, 4 weeks and 8 weeks. Soleus muscle. Four weeks after the process of regeneration started, in control group fiber type distribution was similar to normal SOL without the regeneration. In contrast, in diabetic muscles after eight weeks of regeneration, the percentage of fiber types did not reach values of control group and normal SOL without the process of regeneration. In the diabetic group treated with insulin fiber type distribution after four weeks was similar to these in control group. In diabetic soleus muscles, during all investigated intervals, the fibre cross areas of type I and IIA were significantly smaller than in normal muscle. In diabetic soleus muscle treated with insulin, the fibre cross areas of type I and type IIA were significantly higher than in diabetic muscles after 10 days of treatment, but this effect was diminished after 8 weeks. Extensor digitorum longus After 8 weeks of experiment control muscles were completely regenerated. In diabetic untreated muscles in all observed subgroups, fibers type distribution was changed. After 8 weeks of diabetes the percentage of type I and IIA fibers, which have predominant oxidative metabolism, was greater. The treatment with insulin prevented changes in fiber type distribution during all investigated periods. In diabetic EDL muscle, the cross areas of all fibre types were significantly smaller than in control muscles with an exception of fibre type I after 10 days. In diabetic group treated with insulin the fibre cross areas were significantly higher, than in diabetic group during the all experimental periods. Diabetes mellitus is associated with evident decrease of the regenerative capacity of skeletal muscle. Treatment with insulin in long-term diabetes mellitus can prevent atrophy of regenerative slow and fast skeletal muscles.

Normal aging in humans is associated with progressive decrease in skeletal muscle mass and strength (sarcopenia) which contributes to frailty and falls. The age associated changes in body composition result from lower levels of anabolic hormones, oxidative damage, neuromuscular alterations and a general decrease in muscle protein turnover. In this review we discuss the potential mechanisms and physical activity as prevention and treatment of sarcopenia.

Excitation-transcription coupling, namely the process whereby plasma membrane depolarization leads to gene activation or inactivation, is still a black box for most muscle genes. Muscle regeneration is a useful model system to ask basic questions concerning the triggering signals and the transduction pathways involved in activity-dependent gene regulation. We report ongoing research in our laboratory concerning (1) myosin heavy chain changes in regenerating muscle in the presence and absence of the nerve, as well as changes induced by electrical stimulation, (2) identification of activity response elements in the promoter of a slow myosin light chain gene, and (3) potential approaches to define the transduction pathways induced by neural or electrical activity and implicated in muscle gene regulation.

The regulation of a slow muscle gene, the myosin light chain 1 slow/ventricular gene, has been studied by in vivo transfection into regenerating rat skeletal muscle. Constructs containing portions of the myosin light chain 1 slow/ventricular promoter linked to reporter genes were injected into fast and slow muscles 3 days after muscle injury by bupivacaine injection, and reporter gene activity was analysed after 10 days. We report that a sequence in the 5' flanking region of the myosin light chain 1 slow/ventricular gene is able to direct slow muscle-specific regulation of reporter genes, and that the expression of the transgene, like that of the corresponding endogenous gene, is dependent on intact nerve. This study validates the use of regenerating muscle as a model for studying muscle gene regulation and is the first demonstration of a myosin gene promoter regulated by nerve activity.