Hearing evaluation of school children in Kuwait.

PURPOSE: Hearing level in some children may be abnormal due to conductive or sensorineural causes. In Kuwait, the hearing screening test is not conducted for newborns at public hospitals. We evaluated the hearing level of 159 Kuwaiti school children (age 6-12 years) at the Audiology Clinic of Ahmadi hospital to determine the extent and causes of hearing loss in these children. METHODS: Each child was evaluated by otoscopic examination, tympanogram, acoustic reflex threshold (ART), audiogram, and distortion product oto-acoustic emission (DPOAE). RESULTS: 120 children were found to have normal ear conditions, while 39 children had abnormal results. The abnormalities were 21 children with ear wax, 16 children with secretory otitis media (SOM), and 2 children with sensorineural hearing loss (SNHL). In the children with ear wax, 3 had normal hearing level, while 18 had mild conductive hearing loss in the 250-500 Hz frequency range. In the children with SOM, 3 had normal hearing level, 9 had mild conductive hearing loss in the 250-500 Hz frequency range, while 4 had moderate conductive hearing loss in the 250-2000 Hz frequency range. The children with SNHL had moderate hearing loss in the 4000-8000 Hz frequency range. CONCLUSION: Wax in the ear canal and SOM were the main factors associated with hearing loss in these Kuwaiti school children. It is necessary to establish a national program of school entry hearing screening in Kuwait.

Newborn hearing screening in Kuwait.

OBJECTIVES: The purpose of the study was to detect permanent hearing loss in a universal newborn hearing screening (UNHS) program that included 200 normal ("well baby") newborn children and 15 children at high risk for hearing loss at Ahmadi Hospital in Kuwait. PATIENTS AND METHODS: Distortion product oto-acoustic emissions (DPOAEs) were conducted in the ward on newborns at the age of 2 days. Newborns who did not pass the DPOAE the second time (at the age of 1 month) were evaluated by a brainstem evoked response audiometry (BERA) test within the age of 3 months. If the infant failed the BERA test, the test was repeated at the age of 6 months to confirm any permanent hearing loss. RESULTS: The peak latencies and interpeak intervals of BERA were established for newborns (5-90 days) at our clinic (ms), I: 1.58 + 0.11, III: 4.16 + 0.22, V: 6.70 + 0.24, I-III: 2.57 + 0.19, III-V: 2.59 + 0.24, I-V: 5.12 + 0.26. The incidence of permanent hearing loss was different across nursery levels. In the "well baby" group, 1% had profound sensorineural hearing loss, 1% had severe (70 dBnHL) sensorineural hearing loss, and 98% had normal hearing level. In the "high-risk" group, 26.67% had profound sensorineural hearing loss, 20% had moderate (60 dBnHL) sensorineural hearing loss, and 53.33% had normal hearing level. CONCLUSIONS: The newborn hearing screening program revealed that hearing loss in the "well baby" group is 2%, while in the "high risk" group it is 46.67%. The results warrant the establishment of UNHS programs in Kuwait and other countries in the Middle East to detect permanent hearing loss very early in life and provide appropriate medical treatment.

BERA in children with hearing loss and delayed speech.

OBJECTIVES: The brainstem evoked response audiometry (BERA) is an objective neurophysiological method for the evaluation of the hearing threshold and diagnosing retrocochlear lesions. The aim of the study was to investigate the hearing level in children with suspected hearing loss or pathological speech development. PATIENTS AND METHODS: The BERA diagnostic procedure was applied in 184 children ranging from 1 to 12 years of age at Ahmadi Hospital in Kuwait. RESULTS: We found profound hearing loss (deafness) in 13 children, severe hearing loss in 8 children, moderate hearing loss in 34 children, mild hearing loss in 34 children, and normal hearing level in 95 children. Out of the children suspected for hearing loss, 42% actually had some level (mild-moderate) of hearing loss. Out of the children with delayed speech, 63% had some level (mild-profound) of hearing loss which actually caused the delay in speech development; 37% had normal hearing, but inadequate verbal communication affected their language acquisition and speech development. CONCLUSIONS: These results illustrate the necessity to test children hearing even with the slightest suspicion by the parent or doctor of hearing loss. The results warrant the establishment of a hearing screening test of newborns in Kuwait to provide hearing aid to children with hearing loss.

alpha-Tocopherol modifies lead induced functional changes at murine neuromuscular junction.

Lead impacts neuromuscular junction and might induce skeletal muscle weakness. Antioxidants may prevent toxic actions of lead on muscle. In this study, resting membrane potentials, endplate potentials, miniature endplate potentials (MEPPs) and isometric twitch tensions were recorded to investigate effects of alpha-tocopherol (Vitamin E) on lead induced changes at murine dorsiflexor muscle. Moreover, levels of endplate nicotinic receptors were measured by receptor autoradiography. Forty rats were divided into four groups (lead alone, alpha-tocopherol, lead plus alpha-tocopherol and saline). Lead (1 mg/kg, i.p.), was administered daily for 2 weeks and alpha-tocopherol (100 mg/kg, i.p.) was given daily for 3 weeks. Lead treatment significantly reduced twitch tension (from 4.4+/-0.4 to 2.2+/-0.3 g) and delayed half time of decay. MEPP frequencies and quantal content were also significantly reduced after lead treatment. Pretreatment with alpha-tocopherol reversed twitch tension reduction (4.1+/-0.3 g) and modified lead induced delay in half time of decay. Similarly, alpha-tocopherol modified the negative actions of lead exposure on MEPP frequencies and quantal content. Receptor autoradiographic studies revealed significant increase of nicotinic receptor levels at the endplate region of flexor muscle in lead treated mice. However, animals treated with lead plus alpha-tocopherol showed significantly decreased levels of nicotinic receptors. alpha-Tocopherol appears to protect against lead induced neuromuscular dysfunction. These effects of alpha-tocopherol are possibly mediated via a free radical mechanism or modification of calcium homeostasis.

Effects of alpha-tocopherol on diabetes-induced alterations of synaptic transmission and contractile features in murine dorsiflexor muscle.

Diabetes mellitus affects skeletal muscle and free radicals may be implicated in the manifestation of diabetes complications. The present study investigated effects of alpha-tocopherol on diabetic dorsiflexor muscle via recording resting membrane potentials (RMPs), endplate potentials (EPPs), miniature endplate potentials (MEPPs) and isometric twitch tensions. Forty mice were divided randomly into two groups (n = 20). One group served as control and the other was injected once with streptozotocin (STZ) solution (60 mg/kg, i.p) to induce diabetes. The animals were then divided further into two subgroups (n = 10). Alpha-tocopherol (100 mg/kg, i.p) was administered daily to one control and one diabetic group for 3 weeks prior to recording day. Experiments were conducted 4 weeks following diabetes induction. Isometric twitch tension was measured in anaesthetized mice (2 mg/g urethane, i.p) via a transducer connected to a computer system. Resting membrane potentials and MEPPs were measured by utilizing the intracellular recording method. Compared to control, diabetic mice showed reduced twitch tension (4.4 +/- 0.4 g control vs. 2.5 +/- 0.3 g diabetic) and demonstrated delayed half time of decay. Diabetic flexor muscle also displayed significant reduction in MEPPs frequencies with no changes in RMPs. Alpha-tocopherol reversed tension reduction in diabetic mice (from 2.5 +/- 0.3 to 3.8 +/- 0.4 g), impacted delayed half time of decay and reversed reduction in MEPPs frequencies. Alpha-tocopherol exerts a protective role against diabetes-induced peripheral muscle dysfunction. This effect is probably mediated via a free radical scavenging mechanism or modification of Ca2+ homeostasis.

Effects of ascorbic acid on lead induced alterations of synaptic transmission and contractile features in murine dorsiflexor muscle.

Lead is a common environmental toxin that affects neuromuscular junction and potentially might cause muscle weakness. Antioxidants like ascorbic acid may protect against lead induced myopathy. The present study measured isometric twitch tensions (evoked either directly by muscle stimulation or indirectly by nerve stimulation) to study effects of ascorbic acid on lead induced alterations at murine dorsiflexor skeletal muscle. Resting membrane potentials (RMPs), endplate potentials (EPPs) and miniature endplate potentials (MEPPs) were also recorded. Forty animals were divided into four groups of n = 10 each. (10 control, 10 lead alone, 10 ascorbic acid alone, 10 lead treated plus ascorbic acid). Lead (1 mg/kg) i.p, was administered daily for 2 weeks before the recording day and ascorbic acid (200 mg/kg, i.p) was given daily for 3 weeks prior to the experiment day. Lead treatment reduced twitch tension significantly (from 4.3 +/- 0.5 g to 2.7 +/- 0.2 g) and delayed half time of decay compared to the control. Similarly MEPPs frequencies were reduced following lead treatment. Application of ascorbic acid prevented twitch tension reduction in lead treated mice (3.3 +/- 0.3 g) and reversed lead induced delay in half time of decay. The negative actions of lead treatment on MEPPs frequencies were also modified with ascorbic acid. It appears that ascorbic acid exerts a protective role against lead induced peripheral nerve and muscle dysfunction. This effect of ascorbic acid on lead induced neuromyopathy is probably mediated via a free radical scavenging mechanism or modification of Ca(2+) homeostasis.

The impact of vitamin C on diabetes induced alterations at murine neuromuscular junction.

Physiological functions of skeletal muscle are compromised in diabetes. This may involve free radical mechanisms and may be reversed by antioxidants. We have studied effects of vitamin C on twitch tension, resting membrane potential (RMP) and miniature endplate potentials (MEPPs) frequencies in dorsiflexor muscle of diabetic murine. Forty mice were divided randomly into 2 groups (n = 20 each). One group served as control and the other was injected once with streptozotocin (STZ) solution (60 mg/kg, i.p.) to induce diabetes. The animals were then divided further into two subgroups (n = 10 each). Vitamin C (200 mg/kg, i.p.) was administered daily to one control and one diabetic group for three weeks prior to recording day. Experiments were conducted four weeks following diabetes induction. Isometric twitch tension (evoked directly by muscle stimulation and indirectly by nerve stimulation) was measured in urethane anesthetized (2 mg/g, i.p.) mice via a transducer connected to computer system. Utilizing intracellular recording method, resting membrane potential RMP and MEPPs frequencies were also measured. Compared to control, diabetic mice showed reduced twitch tension (4.2 +/- 0.5 g control versus 2.6 +/- 0.2 g diabetic) and demonstrated delayed half time of decay. Diabetic flexor muscle also displayed significant reduction in MEPPs frequencies with no changes in RMP. Vitamin C reversed tension reduction in diabetic mice (from 2.6 +/- 0.2 g to 3.9 +/- 0.3 g), impacted delayed half time of decay and increased MEPPs frequencies. Vitamin C improves diabetes-induced nerve and muscle dysfunction possibly via a free radical scavenging mechanism.

Reduced potassium currents in old rat CA1 hippocampal neurons.

Potassium currents are an important factor in repolarizing the membrane potential and determining the level of neuronal excitability. We compared potassium currents in CA1 hippocampal neurons dissociated from young (2-3 months old) and old (26-30 months old) Sprague-Dawley rats. Whole-cell patch-clamp techniques were used to measure the delayed rectifier (sustained) and the A-type (transient) potassium currents. The delayed rectifier current was smaller in old (548 +/- 57 pA) than in young (1193 +/- 171 pA) neurons. In the absence of extracellular calcium, the delayed rectifier current was also smaller in old (427 +/- 41 pA) than in young (946 +/- 144 pA) neurons. The cell membrane capacitance was unchanged in old (13.3 +/- 1.2 pF) compared to young (13.6 +/- 1.2 pF). Therefore, the reduction in the delayed rectifier current was not due to a change in membrane surface area. Moreover, activation and inactivation of the delayed rectifier current were unchanged in old compared to young neurons. The slope of the current-voltage relation, however, was smaller in old (B = 5.03) than in young (B = 9.62) neurons. Similarly, the A-current was smaller in old (100 +/- 16 pA) than in young (210 +/- 44 pA) neurons in the presence of extracellular calcium. This reduction of potassium currents could account for the prolongation of action potentials reported previously for old rat CA1 hippocampal neurons. The age-related reduction in potassium current indicates plasticity in neuronal function that can impact communication in the hippocampal neural network during aging.

Early morphological remodeling of neuromuscular junction in a murine model of diabetes.

Although skeletal muscle weakness is documented in diabetes, the time course for its development is not established. The present study examined the dorsiflexor muscle from animals that had been diabetic for 2 wk. Adult male c57BL mice were injected once with streptozotocin (STZ) to induce diabetes (60 mg/kg ip). Two weeks later, resting membrane potential and miniature end-plate potentials were recorded, and electron microscopy was utilized for ultrastructural evaluations. After STZ-induced diabetes, both resting membrane potential and miniature end-plate potentials were reduced. Nerve terminals showed less synaptic vesicles and had degenerated mitochondria. Furthermore, in the intramuscular nerves, disorganization of microtubules and neurofilaments was evidenced. Myelin-like figures were present in intramuscular nerves, neuromuscular junctions, and muscle fibers. At the muscle level, mitochondria were swollen, with disorganization of their cristae, disruption of T tubules, and myofibers with more deposition of glycogen granules. The present results revealed early STZ-induced nerve and muscle alterations. Observed ultrastructural modifications resemble those of motoneuron disorders and aging processes. These changes are possibly related to alterations in Ca(2+) mobilization across muscle membrane. Other mechanisms such as free radical-mediated actions may also be implicated in STZ-induced effects on skeletal muscle.

Cadmium modulates diabetes-induced alterations in murine neuromuscular junction.

Skeletal muscle function is compromised in diabetes mellitus and exposure to heavy metals may further complicate neuromuscular impairments. The present study investigated the effects of cadmium on diabetes induced dorsiflexor muscle dysfunction in C57 BL adult male mice. Forty mice were divided randomly into 2 groups (n=20 each). One group served as control and the other was injected once with i.p. streptozotocin (STZ) solution (60 mg/kg) to induce experimental diabetes. Each group was then divided into two sub-groups (n=10) of which one received 5 mM cadmium. Utilizing intracellular recording method, resting membrane potential (RMP) and miniature endplate potentials (MEPPs) were measured in dorsiflexor muscle obtained from urethane-anaesthetized (2 mg/g, i.p.) four weeks diabetic and matched control mice. Comparative analyses of isometric contractile characteristics of in situ dorsiflexor muscle were also conducted in both groups. In control mice, flexor muscle exposure to 5 mM cadmium for 10 min resulted in significant reduction in MEPPs frequencies and isometric twitch tensions without affecting RMP. In STZ-diabetic mice, the same exposure did not modify resting membrane potential and further decreased MEPPs frequencies and isometric twitch tensions. Current results indicated that cadmium probably via a Ca2+ antagonist and chelating activity at nerve terminals exacerbates diabetes complications.

Progression of visual evoked potential abnormalities in multiple sclerosis and optic neuritis.

To investigate visual signal processing in patients with optic neuritis, suspected multiple sclerosis, confirmed multiple sclerosis (MS), and optic neuritis combined with MS, pattern reversal visual evoked potentials (VEPs) were recorded in patients and normal subjects. The amplitude and latency of the first positive peak, P100 were determined to assess electrical conduction in patients as compared to normal subjects. Suspected MS patients did not differ from normal subjects in peak latencies or amplitudes. The P100 amplitude was reduced in optic neuritis, confirmed MS and optic neuritis combined with confirmed MS. The P100 and N145 latencies were prolonged in optic neuritis patients and confirmed MS patients as compared to normal subjects. The main characteristic of optic neuritis was P100 amplitude reduction, and of confirmed MS was P100 latency delay. There was a progression of the P100 latency delay and of the P100 amplitude decrement in optic neuritis, confirmed MS, and optic neuritis combined with confirmed MS. These results indicate a progression of demyelination in optic neuritis, confirmed MS, and optic neuritis combined with confirmed MS.

Water deprivation reveals early neuromyopathy in diabetic mice.

The effects of water deprivation on peripheral nerve and muscle function were investigated in flexor digitorum superficialis muscle of control and diabetic mice. Twenty mice (30 g average body weight) were injected once with streptozotocin solution (200 mg/kg) to induce experimental diabetes and another 20 mice of similar body weight served as controls. Two weeks later, comparative analyses of in situ muscle isometric contractile characteristics were performed by direct muscle stimulation and indirect nerve stimulation (at 1, 5 and 30 Hz) in urethane-anesthetized (2 mg/g, i.p.) control and diabetic mice. One day prior to the experiments, 10 control and 10 diabetic mice were deprived of water. The study contained four groups: hydrated (H) control, dehydrated (DH) control, H diabetic and DH diabetic. There were no significant differences in synaptic delay or twitch tension between H control and DH control. Comparing H control and H diabetic groups, no differences were noticed in synaptic delay or twitch tension; except at 30 Hz where twitch tension was reduced in H diabetic mice. Significant differences were observed when comparing DH control and DH diabetic mice. DH diabetic showed a significant increase in synaptic delay (from 7.4 to 9.3 ms) and a significant decrease in twitch tension evoked either by indirect nerve or by direct muscle stimulation (from 4.4 g to 1.9 g and from 4.4 g to 2.3 g, respectively). These results revealed that water deprivation enhances diabetes effects at the neuromuscular junction and at the muscle leading to further complication of neuromyopathy.

Potassium current in Drosophila neurons is increased by either dunce mutation or cyclic AMP.

In the Drosophila mutant dunce, short-term memory is deficient and intracellular cyclic adenosine monophosphate (cAMP) concentration is elevated. We examined the effect of increased cAMP concentration on the potassium current. The conventional whole-cell technique was applied to cultured "giant" Drosophila neurons derived from cell-division arrested embryonic neuroblasts. Potassium membrane currents were measured from: 1) control wild-type neurons, 2) wild-type neurons with dibutyryl cAMP and theophylline in the culture media for 2 days (db-cAMP-treated), and 3) dunce neurons. Delayed-rectifier potassium current was greater in both dunce neurons and db-cAMP-treated wild-type neurons than in control wild-type neurons. This result indicates that the neuronal potassium current is increased by the long-term increase of cAMP. Conceivably, altered neuronal excitability in the dunce mutant could disrupt the processing of neural signals necessary for learning and memory.

Modulation of membrane currents by cyclic AMP in cleavage-arrested Drosophila neurons.

A number of Drosophila learning mutants have defective intracellular second-messenger systems. In an effort to develop techniques that will allow direct measurement of the effects of these mutations on whole-cell neuronal membrane currents, the perforated-patch whole-cell (PPWC) technique has been applied to cleavage-arrested cultured embryonic Drosophila neurons. This technique permits the measurement of membrane currents without disturbing the intracellular environment. As a result of the maintenance of the intracellular environment, Drosophila neuron currents are found to be much more stable than when measured using the conventional whole-cell (CWC) patch-clamp technique. Ca2+ channel currents, which typically 'wash out' within a few minutes of the beginning of CWC recording, are stable for the duration of the seal (tens of minutes) when measured using the PPWC technique. Since the learning mutations dunce and rutabaga disrupt cyclic AMP signalling, the action of externally applied dibutyryl cyclic AMP (db-cAMP) and theophylline on Ca2+ and K+ channel currents were studied. db-cAMP and theophylline enhanced the Ba2+ current, carried by Ca2+ channels, but had no effect on the K+ current in the cleavage-arrested neurons. However, the large variability and reduction in density of Ba2+ and K+ currents raise questions about the suitability of using these cleavage-arrested cells as models for Drosophila neurons.

Depolarization reverses age-related decrease of spontaneous transmitter release.

The effect of increasing extracellular Ca concentration on spontaneous transmitter release was studied at soleus nerve terminals of young (10 mo) and old (24 mo) C57BL/6J mice depolarized by high extracellular K concentration ([K]o). By using intracellular recording, miniature end-plate potentials (MEPPs) were first recorded in a normal [K]o Krebs solution. Subsequently, MEPPs were recorded in high [K]o Krebs solutions with four different Ca concentrations: Ca-free/ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and 0.5, 1.5, and 2.5 mM Ca. In both the normal [K]o Krebs and the Ca-free-high [K]o Krebs solutions, MEPP frequency was lower at old than at young nerve terminals. In the three high [K]o Krebs solutions with Ca, MEPP frequency was progressively higher at old than at young nerve terminals with higher Ca concentrations. Periodic oscillations were observed in MEPP frequency of depolarized nerve terminals. The period of oscillation was inversely proportional to spontaneous transmitter release. These results demonstrate that when the nerve terminal is depolarized, permeability of the terminal membrane to Ca increases because of opening of voltage-dependent Ca channels. In the present study resting MEPP frequency was lower at old than at young terminals. On depolarization, MEPP frequency became higher at old than at young terminals. The study demonstrates that voltage-dependent Ca entry increases during aging at the soleus nerve terminal.

Effect of exercise on physiological age-related change at mouse neuromuscular junctions.

To determine the effect of endurance exercise on physiological age-related change at the mouse neuromuscular junction (NMJ), synaptic function was studied for extensor digitorum longus (EDL) and soleus muscles of three C57BL/6J mouse groups, 1) young adult control (YC: 10 months), 2) old control (OC: 20 months), and 3) old mice which exercised (OE: 20 months) since young-adulthood. Electrophysiological properties were studied with intracellular recording techniques. Safety margin was studied by measuring indirect isometric twitch tension in different calcium concentrations. With sedentary aging, EDL and soleus quantal contents increased. Following aging combined with 10 months of exercise, the EDL quantal contents in OE and YC animals were similar. In contrast, soleus quantal content was greater in OE than in YC animals. Determined safety margins were OC greater than YC = OE for EDL, and OC = YC = OE for soleus. This is the first study to indicate that physiological age-related changes at NMJs of EDL and soleus muscles are affected differently by endurance exercise. Exercise prevented all physiological age-related changes in EDL NMJs but not in soleus NMJs, this suggests that EDL changes are associated with inactivity during aging, while soleus changes are "fundamental" age changes.

Aging increases calcium influx at motor nerve terminal.

To determine whether increased transmitter release from soleus nerve terminals of old C57BL/6J mice is caused by an altered Ca2+ regulation, the time course of post-tetanic potentiation of miniature endplate potential (MEPP) frequency was used as an indicator of the kinetics of Ca2+ metabolism in young (10 months) and old (24 months) mice. Post-tetanic potentiation properties were studied in either (1) 0.2 mM Ca2+, 5.0 mM Mg2+ Krebs; or (2) Ca2(+)-free/EGTA Krebs to eliminate Ca2+ influx, and thereby isolated Ca2+ buffering. In the 0.2 mM Ca2+ Krebs, the time constants of decay of augmentation (TA) and potentiation (TP) were longer in old (TA = 10.3 +/- 1.0 sec, TP = 195.3 +/- 5.4 sec) than in young (TA = 7.0 +/- 0.7 sec, TP = 78.8 +/- 6.6 sec) nerve terminals. Evoked transmitter release was measured in 0.4 mM Ca2+, 2.75 mM Mg2+ Krebs. Quantal content of the endplate potential was positively correlated with TA (r = 0.95) and with TP (r = 0.98). In the Ca2(+)-free/EGTA Krebs, there was no difference in post-tetanic potentiation properties between young and old terminals. These results suggest that Ca2+ influx into the soleus nerve terminal increases with aging. This may explain, at least in part, the increased quantal content observed at old terminals.