Grzegorz Witkowski

Fascicles of the adult human Achilles tendon - an anatomical study.

The Achilles or calcaneal tendon is the structural base for the biomechanical work of the ankle joint. The purpose of this study is to describe the internal structure of the human Achilles tendon. The anatomy of the Achilles tendon has been described in lower mammals in which it has three parts which can be dissected from its beginning to the insertion onto the calcaneus. The partial ruptures of each part suggest that the human Achilles tendon may also be composed of parts. The Achilles tendon is one of the most commonly torn tendons in the human body. Each segment of the Achilles tendon described by us can be ruptured separately, which can cause a partial dysfunction in flexion of the ankle joint as observed in clinical practice. We dissected 20 Achilles tendons previously fixed in 10% formaldehyde and 20 fresh-frozen Achilles tendons, paying particular attention to the relationship between the lateral and medial heads of the gastrocnemius and the soleus muscles. The layer-by-layer method and a microscope were used in our study. We found that the medial group of fibers from the medial head of the gastrocnemius muscle constitutes the posterior layer of the tendon. The lateral border of the tendon is composed of the fibers from the lateral part of the medial head of the gastrocnemius muscle. The fibers from the lateral head of the gastrocnemius muscle constitute the anterior layer of the Achilles tendon. The fibers from the soleus muscle are located in the anteromedial part of the Achilles tendon. Our findings are supported by clinical descriptions and observations of the partial rupture of the Achilles tendon.

D1 DOPAMINERGIC CONTROL OF G PROTEIN-DEPENDENT INWARD RECTIFIER K+ (GIRK)-LIKE CHANNEL CURRENT IN PYRAMIDAL NEURONS OF THE MEDIAL PREFRONTAL CORTEX

Pyramidal neurons of the medial prefrontal cortex (mPFC) exhibit dopamine-dependent prolonged depolarization, which may lead to persistent activity. Persistent activation of prefrontal cortex neurons has been proposed to underlie the working memory process. The purpose of our study was to test the hypothesis that activation of D(1) dopamine receptors leads to inhibition of G protein-dependent inward rectifier K(+) (GIRK) channels, thereby supporting the prolonged depolarization of mPFC pyramidal neurons. Experiments were performed on 3-week-old rats. GIRK-like channel currents recorded from pyramidal neurons showed the following properties at -75 mV: open probability (NPo), 2.5+/-0.3 x 10(-3); mean open time, 0.53+/-0.05 ms; and conductance, 29.9+/-1.6 pS (n=60). The channel currents were strongly inward-rectified. GIRK channel currents were reversibly inhibited by the D(1) agonists SKF 38393 (10 microM) and SKF 81297 (10 microM). This inhibition was abolished by prior application of a dopamine receptor antagonist and by application of the membrane-permeable protein kinase C inhibitors chelerythrine chloride (3 microM) and calphostin C (10 microM). It was also found that the application of D(1) dopamine receptor agonists or GIRK channel inhibitors evoked depolarization of mPFC pyramidal neurons in rats. Moreover, prior application of a GIRK channel blocker eliminated the depolarizing effect of D(1) agonists. We conclude that activation of D(1) dopamine receptors may lead to inhibition of GIRK channel currents that may, in turn, lead to the prolonged depolarization of mPFC pyramidal neurons in juvenile rats.

Opioid μ receptor activation inhibits sodium currents in prefrontal cortical neurons via a protein kinase A- and C-dependent mechanism

Opioid transmission in the medial prefrontal cortex is involved in mood regulation and is altered by drug dependency. However, the mechanism by which ionic channels in cortical neurons are controlled by mu opioid receptors has not been elucidated. In this study, the effect of mu opioid receptor activation on voltage-dependent Na(+) currents was assessed in medial prefrontal cortical neurons. In 66 out of 98 nonpyramidal neurons, the application of 1 microM of DAMGO ([D-Ala(2), N-Me-Phe(4), Gly(5)-OL]-enkephalin), a specific mu receptor agonist, caused a decrease in the Na(+) current amplitude to approximately 79% of that observed in controls (half blocking concentration = 0.094 microM). Moreover, DAMGO decreased the maximum current activation rate, prolonged its time-dependent inactivation, and shifted the half inactivation voltage from -63.4 mV to -71.5 mV. DAMGO prolonged the time constant of recovery from inactivation from 5.4 ms to 7.4 ms. The DAMGO-evoked inhibition of Na(+) current was attenuated when GDP-beta-S (0.4 mM, Guanosine 5-[beta-thio]diphosphate trilithium salt) was included in the intracellular solution. Inhibitors of kinase A and C greatly attenuated the DAMGO-induced inhibition, while adenylyl cyclase and kinase C activators mirrored the DAMGO inhibitory effect. Na(+) currents in pyramidal neurons were insensitive to DAMGO. We conclude that the activation of mu opioid receptors inhibits the voltage-dependent Na(+) currents expressed in nonpyramidal neurons of the medial prefrontal cortex, and that kinases A and C are involved in this inhibitory pathway.

Expression and kinetic properties of Na(+) currents in rat cardiac dorsal root ganglion neurons.

The expression and properties of voltage-gated Na(+) currents in cardiac dorsal root ganglion (DRG) neurons were assessed in this study. Cardiac DRG neurons were labelled by injecting the Fast Blue fluorescent tracer into the pericardium. Recordings were performed from 138 cells. Voltage-dependent Na(+) currents were found in 115 neurons. There were 109 neurons in which both tetrodotoxin-sensitive (TTX-S, blocked by 1 microM of TTX) and tetrodotoxin-resistant (TTX-R, insensitive to 1 microM of TTX) Na(+) currents were present. Five cells expressed TTX-R current only and one cell only the TTX-S current. The kinetic properties of Na(+) currents and action potential waveform parameters were measured in neurons with cell membrane capacitance ranging from 15 to 75 pF. The densities of TTX-R (110.0 pA/pF) and TTX-S (126.1 pA/pF) currents were not significantly different. Current threshold was significantly higher for TTX-R (-34 mV) than for TTX-S (-40.4 mV) currents. V(1/2) of activation for TTX-S current (-19.6 mV) was significantly more negative than for TTX-R current (-9.2 mV), but k factors did not differ significantly. V(1/2) and the k constant for inactivation for TTX-S currents were -35.6 and -5.7 mV, respectively. These values were significantly lower than those recorded for TTX-R current for which V(1/2) and k were -62.3 and -7.7 mV, respectively. The action potential threshold was lower, the 10-90% rise time and potential width were shorter before than after the application of TTX. Based on this we drew the conclusion that action potential recorded before adding tetrodotoxin was mainly TTX-S current dependent, while the action potential recorded after the application of toxin was TTX-R current dependent. We also found 23 cells with mean membrane capacitance ranging from 12 to 35 pF (the smallest labelled DRG cells found in this study) that did not express the Na(+) current. The function of these cells is unclear. We conclude that the overwhelming majority of cardiac dorsal root ganglion neurons in which voltage-dependent Na(+) currents were present, exhibited both TTX-S and TTX-R Na(+) currents with remarkably similar expression and kinetic properties.

Voltage-dependent k+ currents in rat cardiac dorsal root ganglion neurons

We have assessed the expression and kinetics of voltage-gated K(+) currents in cardiac dorsal root ganglion (DRG) neurons in rats. The neurons were labelled by prior injection of a fluorescent tracer into the pericardial sack. Ninety-nine neurons were labelled: 24% small (diameter<30 microm), 66% medium-sized (diameter 30 microm>.48 microm) and 10% large (>48 microm) neurons. Current recordings were performed in small and medium-sized neurons. The kinetic and pharmacological properties of K(+) currents recorded in these two groups of neurons were identical and the results obtained from these neurons were pooled. Three types of K(+) currents were identified:a) I(As), slowly activating and slowly time-dependently inactivating current, with V(1/2) of activation -18 mV and current density at +30 mV equal to 164 pA/pF, V(1/2) of inactivation at -84 mV. b) I(Af) current, fast activating and fast time-dependently inactivating current, with V(1/2) of activation at two mV and current density at +30 mV equal to 180 pA/pF, V(1/2) of inactivation at -26 mV. At resting membrane potential I(As) was inactivated, whilst I(Af), available for activation. The I(As) currents recovered faster from inactivation than I(Af) current. 4-Aminopiridyne (4-AP) (10 mM) and tetraethylammonium (TEA) (100 mM) produced 98% and 92% reductions of I(Af) current, respectively and 27% and 66% of I(As) current, respectively. c) The I(K) current that did not inactivate over time. Its V(1/2) of activation was -11 mV and its current density equaled 67 pA/pF. This current was inhibited by 95% (100 mM) TEA, whilst 4-AP (10 mM) produced its 23% reduction. All three K(+) current components (I(As), I(Af) and I(K)) were present in every small and medium-sized cardiac DRG neuron. We suggest that at hyperpolarized membrane potentials the fast reactivating I(As) current limits the action potential firing rate of cardiac DRG neurons. At depolarised membrane potentials the I(Af) K(+) current, the reactivation of which is very slow, does not oppose the firing rate of cardiac DRG neurons.

Voltage-dependent Ca2+ currents in rat cardiac dorsal root ganglion neurons

This study presents the kinetic and pharmacological properties of voltage-gated Ca(2+) currents in anatomically defined cardiac dorsal root ganglion (DRG) neurons in rats. The neurons were labelled by prior injection of fluorescent tracer Fast Blue into the pericardial sack. There were three distinct groups of neurons with respect to cell size: small (27% of total; cell capacitance <30 pF), medium (65% of total; capacitance 30-80 pF) and large neurons (8% of total; capacitance >80 pF). The properties of Ca(2+) currents were tested in small and medium-sized neurons. In large neurons currents could not be adequately controlled and were not analysed. Ca(2+) currents did not completely inactivate during 100 ms depolarising voltage steps. The activation thresholds in small (-36.9+/-1.3 mV) and medium (-39.0+/-1.3 mV) size neurons were similar. Current densities were 105.8 pA/pF in small and 97.4 pA/pF in large neurons and also did not differ. The kinetic properties of activation and inactivation did not differ between small and medium-sized cardiac DRG neurons. At membrane potentials between -50 and -60 mV (the expected resting membrane potential in these neurons) 55 to 70% of Ca(2+) currents in small and medium-sized neurons were available for activation. Both, small and medium-sized neurons expressed similar proportions of L (7.5%), N (25%) and P/Q (36%) type Ca(2+) currents. We conclude that small and medium-sized cardiac DRG neurons are homogeneous with respect to the expression and properties of voltage-gated Ca(2+) currents. Voltage-gated Ca(2+) currents probably play an important role in action potential generation in cardiac DRG neurons due to their availability for activation at resting membrane potential, their high density and voltage threshold close to the threshold for voltage-gated Na(+) currents.

Effects of ATP and GTP on voltage-gated K+ currents in glandular and muscular sympathetic neurons.

This study assesses the effects of ATP and GTP on the kinetic properties of voltage-gated K+ currents in anatomically identified postganglionic sympathetic neurons innervating the submandibular gland and the masseter muscle in rats. Three types of K+ currents were isolated: the I(Af) steady-state inactivating at more hyperpolarized potentials, I(As) steady-state inactivating at less hyperpolarized potentials than I(Af) and the I(K) current independent of membrane potential. The kinetic properties of these currents were tested in neurons with ATP (4 mM) and GTP (0.5 mM) or without ATP and GTP in the intracellular solution. In glandular and muscular neurons in the absence of ATP and GTP in the intracellular solution, the current density of I(Af) was significantly larger (142 pA/pF and 166 pA/pF, respectively) comparing to cells with ATP and GTP (96 pA/pF and 100 pA/pF, respectively). The I(As) was larger only in glandular neurons (52 pA/pF vs. 37 pA/pF).Conversely, I(K) current density was smaller in glandular and muscular neurons without ATP and GTP (17 pA/pF and 31 pA/pF, respectively) comparing to cells with ATP and GTP (57 pA/pF and 58 pA/pF, respectively). In glandular (15.5 nA/ms vs. 6.9 nA/ms) and muscular (10.9 nA/ms vs. 7.5 nA/ms) neurons, the I(Af) activated faster in the absence of ATP and GTP. Half inactivation voltage of I(Af) in glandular (-110.0 mV vs. -119.7 mV) and muscular (-108.4 vs. -117.3 mV) neurons was shifted towards depolarization in the absence of ATP and GTP. We suggest that the kinetic properties of K+ currents in glandular and muscular sympathetic neurons change markedly in the absence of ATP and GTP in the cytoplasm. Effectiveness of steady-state inactivated currents (I(Af) and I(AS)) increased, while effectiveness of steady-state noninactivated currents decreased in the absence of ATP and GTP. The effects were more pronounced in glandular than in muscular neurons.

Midbrain and bilateral paramedian thalamic stroke due to artery of Percheron occlusion.

INTRODUCTION Bilateral thalamic strokes are rare manifestations of posterior circulation infarcts. Usually the etiology is cardioembolic or small vessel disease combined with individual anatomical predisposition. The symptoms include a variety of neurological deficits depending on thalamic structure involvement, such as paresthesias or numbness, hemiparesis with increased reflexes and Babinski sign, third cranial nerve palsy, speech and cognition disturbance, memory impairment and stupor. Neuroimaging usually reveals ischemic loci in adequate thalamic nuclei. CASE PRESENTATION We report a case of 61-year-old man, active smoker (25/per day, 50 pack-years) with untreated hypertension who presented at admission consciousness impairment (Glasgow Coma Scale score 9 points), left pupil dilatation without reaction to light, left eye deviation downwards and outwards, vertical gaze paralysis and left-sided hemiplegia. Initial brain computed tomography (CT) was normal. Brain magnetic resonance with diffusion weighted imaging and fluid attenuation inversion recovery sequences (MR DWI/FLAIR) performed on admission showed ischemic changes in bilateral thalami, which were confirmed in routine MRI. Thrombosis of basilar artery and cerebral venous was excluded in CT angiography. Further diagnostic assessment revealed hyperlipidemia, paroxysmal atrial fibrillation and renal cancer with hepatic metastases. CONCLUSION Bilateral thalamic stroke due to artery of Percheron occlusion is a rare presentation of stroke, which can be overlooked in routine CT scan. If diagnosed, it requires further evaluation for stroke risk factors, especially cardiovascular disorders associated with increased embolic risk.

An unusual presentation of Listeria monocytogenes rhombencephalitis

We describe a case of 52-year-old woman with a medical history of Crohns disease presented abrupt fever, asymmetrical multiple cranial nerve palsies and focal neurological symptoms localized to the brainstem. The patient was initially diagnosed with ischaemic stroke, because of acute clinical course and results of neuroimaging. Cerebrospinal fluid analysis revealed mild infection with negative Gram staining and culture. Final diagnosis of Listeria monocytogenes brainstem infection (rhombencephalitis) was set up on the basis of further clinical course and positive blood cultures. Listerial rhombencephalitis should be kept in mind in immunocompromised adult patients who develop fever, asymmetrical multiple cranial nerve palsies and focal neurological symptoms localized to the brainstem even without typical neuroimaging, cerebrospinal fluid findings and negative cultures. Early diagnosis and adequate antibiotic treatment is of crucial importance.

Enzymatic replacement therapy in patients with late-onset Pompe disease – 6-Year follow up

INTRODUCTION Late-onset Pompe disease (LOPD) is a progressive metabolic myopathy, affecting skeletal muscles, which, if untreated, leads to disability and/or respiratory failure. The enzyme replacement therapy (ERT) improves muscle strength and respiratory function and prevents disease progression. We present a 6-year follow-up of 5 patients with LOPD treated with ERT. METHODS Five patients with LOPD received ERT: two started treatment in 2008, other two in 2010 and one in 2011. All patients received recombinant human alpha-glucosidase in dose 20mg/kg intravenously every two weeks. Physical performance was assessed in 6-minute walk test (6MWT) and spirometry was performed to examine FVC and FEV1. Liver enzymes, CK levels were also assessed. RESULTS The walking distance in 6MWT increased by average 16.9±2.26% in the first three years of treatment. Similar changes were detected in spirometry: the most significant FVC increase was observed in two patients with the highest FVC values before treatment, which increased to normal values adjusted for age and sex in three years of treatment, that is by 28% and 34%. In two other patients FVC reached 88% and 76% of predicted values. ERT also improved the liver and muscle enzymes levels. CONCLUSION The improvements of exercise tolerance and FVC were observed in all patients in the first three years of treatment and were the most pronounced in the longest-treated patients and with the least severe neurological and respiratory symptoms. Our research suggests that early start of the ERT results in higher improvement of respiratory and ambulation functions.