Regulation of cytokine expression in human plasmacytoid dendritic cells by prostaglandin I2 analogues.

Plasmacytoid dendritic cells (pDCs) are critical in controlling adaptive immunity, but the mechanisms governing cytokine expression remain incompletely defined. Analogues of prostaglandin (PG)I(2), such as iloprost, can modulate functions of myeloid dendritic cells, but their involvement in the regulation of human pDCs remains unknown. To this end, the regulatory role of PGI(2) analogues on cytokine expression in pDCs was investigated. Circulating pDCs were magnetically sorted with BDCA-4 cell isolation kits from human peripheral blood mononuclear cells and treated with varying concentrations of iloprost with or without the addition of Toll-like receptor agonists, or an I prostanoid (IP) receptor antagonist, CAY10449. The levels of tumour necrosis factor (TNF)-alpha, interferon (IFN)-alpha and interleukin (IL)-10 were measured by ELISA. Iloprost induced IL-10 expression, but suppressed CpG oligodeoxynucleotide- (or imiquimod-) induced TNF-alpha and IFN-alpha production in pDCs. This effect was reversed by the addition of CAY10449. Forskolin, a cyclic adenosine monophosphate activator, conferred a similar modulating effect to that noted in iloprost-treated pDCs, although a higher concentration of forskolin was required to exert the same effect. Iloprost enhanced interleukin-10 and suppressed Toll-like receptor-mediated tumour necrosis factor-alpha and interferon-alpha production of human plasmacytoid dendritic cells via the I prostanoid receptor and, in part, the cyclic adenosine monophosphate pathway.

Effects of formoterol and salmeterol on the production of Th1- and Th2-related chemokines by monocytes and bronchial epithelial cells.

It is unknown whether formoterol and salmeterol, two long-acting beta(2)-adrenoreceptor agonists, have regulatory functions in the production of T-helper cell (Th) type 2- and Th1-related chemokines by monocytes and bronchial epithelial cells. In the present study, the effects of formoterol and salmeterol on lipopolysaccharide (LPS)-induced expression of the Th2-related chemokine macrophage-derived chemokine (MDC; CCL22) and the Th1-related chemokine interferon-gamma-inducible protein (IP)-10 (CXCL10) were investigated in a monocytic cell line, THP-1, and in human primary monocytes. In addition, their effects on the expression of the Th2-related chemokine thymus- and activation-regulated chemokine (TARC; CCL17) were evaluated in an epithelial cell line, BEAS-2B. Formoterol enhanced MDC but suppressed IP-10 production in monocytes induced by LPS. Higher doses of salmeterol were required to enhance LPS-induced MDC expression in THP-1 cells. Formoterol and salmeterol could significantly suppress TARC expression in BEAS-2B cells. These effects could be reversed by a selective beta(2)-adrenoreceptor antagonist, ICI-118551. Formoterol- and LPS-induced MDC expression was inhibited by budesonide. Both long-acting beta(2)-adrenoreceptor agonists suppressed thymus- and activation-regulated chemokine expression in bronchial epithelial cells mediated via beta(2)-adrenoreceptors. Formoterol at physiological concentrations could suppress lipopolysaccharide-induced T-helper cell type 1-related chemokine (interferon-gamma-inducible protein-10) but enhance T-helper cell type 2-related chemokine (macrophage-derived chemokine) expression in human monocytes. Long-acting beta(2)-adrenoreceptor agonists may increase T-helper cell type 2-related chemokine expression in monocytes and T-helper cell type 2 recruitment and, therefore, long-acting beta(2)-adrenoreceptor agonist monotherapy may not be an appropriate therapeutic option for asthma.

Superior mesenteric artery syndrome associated with hereditary motor and sensory neuropathy type II--a case report.

A 14-year-old girl with superior mesenteric artery (SMA) syndrome associated with hereditary motor and sensory neuropathy (HMSN) type II is reported. The initial presentations of HMSN type II were developmental delay and gait disturbance at 2 years of age. All deep tendon reflexes were absent. Nerve conduction velocities and left sural nerve biopsy all revealed axonal changes. Recently, she suffered from intermittent bilious vomiting and epigastralgia for 6 months. That caused body weight loss from 40 kg to 28 kg. Abdominal echography showed narrowed superior mesenteric artery angle. Upper gastrointestinal series revealed obstruction of third portion of duodenum. Accordingly, SMA syndrome was diagnosed. To the best of our knowledge, this case is the first report of SMA with HMSN type II in the world. When a child with chronic neurological disease presents with intermittent vomiting, SMA should be considered as a disease entity of differential diagnosis.

Preparation and properties of SBS-g-DMAEMA copolymer membrane by ultraviolet radiation.

A styrene-butadiene-styrene triblock copolymer (SBS) membrane was prepared by solvent casting. Grafting of dimethyl amino ethyl methacrylate (DMAEMA) to this SBS membrane was subsequently conducted by ultraviolet radiation-induced graft copolymerization without degassing to obtain a SBS-g-DMAEMA copolymer membrane. The graft copolymer was characterized by infrared spectroscopy and scanning electron microscopy. The degree of grafting and the mechanical properties of SBS and SBS-g-DMAEMA were measured. Contact angle, water content, and protein absorption of fibrinogen and albumin experiments were also performed to evaluate the biocompatibility of SBS-g-DMAEMA graft copolymer membranes. It was found that the degree of grafting was related to the irradiation time, DMAEMA concentration, and temperature. The tensile strength of the SBS-g-DMAEMA membrane increased with an increase in the degree of grafting. By using Kaelble's equation and the contact angle data, the surface tension of SBS-g-DMAEMA was determined. It was found that with an increase in the degree of grafting, the surface tension and water content of SBS-g-DMAEMA membrane increased, whereas the contact angle decreased. The amount of absorption of albumin and fibrinogen decreased with an increase in amount of grafting. However, there was a minimum for the adsorption of proteins in the SBS-g-DMAEMA membrane.

Respiratory distress and selective muscle involvement in central core disease: report of a case.

A 10-year-old girl who had suffered life-threatening respiratory distress in late infancy and mild muscle weakness in the extremities was found to have central core disease (CCD). Her developmental milestones were markedly delayed and she became ambulant at the age of 6 years. On physical examination, she had a high-arched palate, nasal tone vocalization, a positive Gowers' sign, waddling gait, and decreased deep tendon reflexes. On magnetic resonance imaging (MRI) examination, the thigh, calf, arm, and paravertebral muscles were selectively involved, especially in the paravertebral, biceps brachii, gluteus maximus and medius, vastus lateralis and medialis, sartorius, adductor magnus, tibialis anterior, soleus, lateral head of gastrocnemius, and peroneus longus muscles. Two biopsy specimens from the left biceps brachii and rectus femoris muscles showed the common histochemical findings of CCD, including type 1 fiber atrophy and predominance, and core structures. Proliferation of adipose tissue was only present in the biceps brachii muscle. The patient improved clinically in muscle strength after three year follow-up. Muscle MRI may provide a non-invasive evaluation of gross muscle involvement in CCD.

Human lung fibroblasts express multiple means for enhanced activity of bradykinin receptor pathways.

Human lung fibroblasts represent important targets for the biologic activities of bradykinin (BK). We have identified multiple mechanisms in these cells which may extend their potential for BK receptor responsiveness, particularly with regard to generation of arachidonate metabolites. These fibroblasts can constitutively express B2 and B1 BK receptors concurrently, both coupled to the pathway for arachidonate metabolism resulting in generation of PGE2 and the potent vasoactive lipid mediator Thromboxane A2. Although expression patterns for B2 and B1 receptors have classically been regarded as 'constitutive' and 'inducible', respectively, we demonstrate that in human lung fibroblasts both can be expressed spontaneously at equivalent biologic activity levels without selective induction by other mediators. Concurrent B2/B1 receptor expression extends the scope of fibroblast response potential to both BK and des-Arg9-BK in the same time frame. We have identified additional short-term and long-term cellular events, involving both protein kinase pathways through which BK receptors act and those which act upon BK receptors, that result in enhanced BK receptor response potential. These properties of BK receptors may affect whether fibroblast behaviors maintain controlled activities of normal homeostasis or foster escalating cellular responses which may influence the progression of certain human disease states.

Protein kinases A and C rapidly modulate expression of human lung fibroblast B2 bradykinin receptor affinity forms.

WI-38 and IMR90 human lung fibroblasts express B2 receptors for the peptide mediator bradykinin. These G-protein-coupled receptors which control cell growth, protein synthesis, and prostaglandin E2 (PGE2) production occur in three affinity forms, high (H, KD 440 pM), intermediate (I, KD 5.6 nM), and low (L, KD 42 nM). Utilizing specific monoclonal antireceptor antibodies which are able to distinguish among these B2 bradykinin receptor forms, we demonstrate regulated differential enhancement of their expression in fibroblasts. Activation of cellular second messenger regulatory pathways based on protein kinase C or protein kinase A drives B2 receptor affinity form expression in opposite directions, both of which are relevant to the levels of human bradykinin generation in vivo in the tissues of origin for these fibroblasts. On a spontaneous basis WI-38 human lung fibroblasts most frequently express the L form alone or the I+L forms concurrently. Activation of protein kinase C augments expression of both I and L affinity receptors within 30 min, increasing receptor number and enhancing PGE2 production. In contrast, activation of protein kinase A by 8-bromo-cAMP or forskolin enhances receptor expression and PGE2 production instead at the I to H types of affinity forms within 30 min. The effects of both kinase systems are blocked by serine/threonine (Ser/Thr) protein kinase inhibitors, indicating a role for phosphorylation at Ser or Thr residues in determining the cellular expression of bradykinin B2 receptor affinity forms. An increase in immunoprecipitable I form bradykinin receptors is detectable within 20 to 30 min after activation of either protein kinase C or protein kinase A. This time frame emphasizes the ability of human fibroblasts for rapid mobilization of B2 receptor affinity forms. Regulated expression of this repertoire of bradykinin B2 receptors at the level of receptor number and concurrent activity allows fibroblasts a sensitive means to adjust their responses to their cellular environment utilizing Ser/Thr phosphorylation events.

[Clinical significance of dystrophin positive fibers in Duchenne muscular dystrophy].

Dystrophin is a protein product of the Xp 21 gene which is defective in patients with Duchenne muscular dystrophy (DMD). In immunohistochemical staining of dystrophin, the majority of DMD muscle fibers show negative staining. Nevertheless, biopsy specimens from DMD patients labeled with many different antibodies may show a small number of fibers which are clearly dystrophin positive. This very small percentage of dystrophin-positive fibers (DPF) probably represents somatic reversion, suppression of the DMD gene mutation or alternating splicing of dystrophin mRNA. To determine the significance of isolated DPF in muscle specimens of DMD patients, we examined 30 DMD muscle specimens, aged 4 years to 16 years, by the use of monoclonal antibody against the C-terminal region of dystrophin. Additionally, muscle specimens from 2 normal human controls and 2 mice with X-linked muscular dystrophy were used for positive and negative controls, respectively. Muscle specimens from DMD patients and mdx mice showed almost totally negative dystrophin staining in most of their muscle fibers, but in 20 patients, three was a trace of isolated DPF ranging from 0.06% to 0.77%. DMD patients with no isolated DPF seemed to have higher functional disability. In the whole group of 30 patients, a significant negative correlation was found between the abundance of DPF and clinical functional grading (r = -0.85, p < 0.0001, based on linear regression). It is suggested that even the very low concentrations of dystrophin found in DMD patients may have a favorable functional significance.

MELAS syndrome: correlation between clinical features and molecular genetic analysis.

The clinical manifestations and mitochondrial DNA (mtDNA) mutations in a Taiwanese family with a female proband exhibiting mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes syndrome are reported. Clinically, the proband had a stroke-like episode with right hemiparesis, hemianopsia and mental dysfunction as well as short stature, hearing impairments, and elevated lactate levels. Brain magnetic resonance images showed multiple increased signal intensities over the left frontal, parietal and temporal areas. There were no ragged-red fibers, but paracrystalline inclusion bodies were shown in the muscle biopsies under electron microscopic examination. A deficiency of NADH-CoQ reductase was also found in biochemical studies of the muscles. The family survey revealed no abnormal findings except for headache and episodic vomiting in her mother. The molecular analysis of mtDNA disclosed a mutation from A to G at the nucleotide pair 3243 of the mitochondrial transfer RNA(Leu) gene in the blood, hair follicles and/or muscle of the maternal relatives. A characteristic finding of the MELAS family is variation of percentage of mutated mtDNA in various tissues and individuals. However, a higher proportion of mutated mtDNA was noted in the proband than that in the asymptomatic or oligosymptomatic family members. From the data, the variable clinical phenotypes in this MELAS family can be explained at least partly, by the different proportions of mutant mtDNA in the target tissues of the proband and maternal relatives.

Leigh disease (subacute necrotizing encephalomyelopathy): report of one case.

A case is reported of a 16-month-old girl who presented with generalized hypotonia, ptosis and persistent low grade fever after a previous pneumonia. Brain CT and MRI showed symmetric necrotizing lesions in the basal ganglia, substantia nigra and periaqueduct area. Lactate and pyruvate levels were elevated in both the blood and cerebrospinal fluid. Biopsy of the rectus femoris muscle for electron microscopic examination revealed some distortion of the mitochondrial cristae. Biochemical study showed normal respiratory chain enzymes. Leigh disease was considered from the neuroradiological findings and morphological investigations.

Mental retardation in congenital nonprogressive myopathy with uniform type 1 fibers.

A 12-year-old girl with proximal muscle weakness and delayed psychomotor development from early infancy is presented. She had a myopathic face, high-arched palate, nasal tone vocalization, positive Gowers' sign, waddling gait and decreased deep tendon reflexes. Her IQ was 40 (PIQ = 39, VIQ = 51). The serum creatine kinase level and peripheral nerve conduction velocity as well as electrocardiogram were normal. The electromyogram showed myopathic changes. Magnetic resonance imaging (MRI) of thigh muscles revealed fatty infiltration of all muscle groups, the hypertrophic biceps femoris and semimembranous muscles being spared. A biopsy specimen from the left biceps brachii muscle revealed small caliber fibers, increased variability of fiber size and uniformity of type 1 fibers (greater than 99%). This case was diagnosed as having congenital nonprogressive myopathy with uniform type 1 fibers, and had a non-deteriorating clinical course as in most congenital nonprogressive myopathy cases.

Neonatal myotubular myopathy with respiratory distress: report of a case.

The case of a 2-day-old male full-term newborn with myotubular (centronuclear) myopathy is reported. He presented with generalized hypotonia and muscle weakness, swallowing disturbance, and respiratory distress at birth. He had a typical myopathic face, high-arched palate, funnel chest, and mild bilateral ptosis. Deep tendon reflexes were absent. Serum creatine kinase was normal. The histologic examination of the muscle biopsy over the right rectus femoris muscle revealed an increased number of fibers with centrally placed nuclei, type 1 fiber predominance, type 1 fiber atrophy, and a peripheral halo in the sarcoplasm on NADH-TR staining. On electron microscopy, central nuclei were separated by strands of glycogen and mitochondria. His muscle strength showed clinical improvement at a 14-month follow-up. This case illustrates the need for a muscle biopsy and histochemical staining and/or electron microscopic investigation for a proper diagnosis in hypotonic newborns with respiratory distress.

Minimal change myopathy: report of a case.

A young boy, aged 5 years 7 months, presented with generalized hypotonia and proximal muscle weakness, and had exhibited delayed motor milestones since birth, He showed talipes planovalgus, a myopathic face, nasal tone vocalization, positive Gowers' sign and decreased tendon reflexes, but there was no intellectual impairment or seizure. The serum creatine kinase level and peripheral nerve conduction velocity, as well as the electromyogram and electrocardiogram, were within normal limits. A biopsy specimen from the left biceps brachii muscle revealed minimal nonspecific changes and mild variations in fiber size with an increased number of undifferentiated type 2C fibers, but no subcellular abnormalities were found on either the histochemical or electron microscopic examinations. The patient was diagnosed as having minimal change myopathy and improved clinically in muscle strength after one year of follow-up.

Congenital fiber type disproportion: report of one case.

Congenital fiber type disproportion (CFTD) is described clinically as muscle weakness and hypotonia with delayed motor development, usually from infancy. Muscle biopsy reveals that type 1 fibers predominate and smaller than type 2 fibers by a margin greater than 12% of the diameter of the type 2 fibers. There are no other subcellular abnormalities, and generally prognosis is good. The CFTD case is a six-month-old girl who manifested clinically as a floppy infant. A biopsied specimen from the left biceps brachii muscle revealed type 1 fiber predominance and hypoplasia with an increased number of undifferentiated type 2C fibers. Electromyogram, nerve conduction velocity and serum creatine kinase level were normal. The child learned to walk without assistance at 1 year 7 month old. Now 2-years and 10-months old, she can climb stairs without difficulty, although she cannot run fast.

Clinical observations and virological study of aseptic meningitis in the Kaohsiung area.

During 1988, an endemic outbreak of aseptic meningitis was noted in the Kaohsiung area. Throughout the year, a total of 89 cases were identified by cerebrospinal fluid (CSF) examination at the Pediatric Department of Kaohsiung Medical College. The peak incidence was from June to October. Scattered cases still occurred during November and December. The male to female ratio was 1.7:1 and the age distribution ranged from 1 month to 15 years old. Two peaks of age distribution were observed; one in infancy and the other in the 4-7 year old age group. Most of them exhibited fever (94.4%), headache (68.9%), and vomiting (68.5%). Other associated symptoms and signs included neck stiffness, sore throat, cough, Brudzinski's sign, abdominal pain, seizure, dizziness, rhinorrhea, diarrhea, Kernig's sign, skin rash, hyperemic conjunctiva, apnea, and oral ulcers. Most of them had CSF white blood cell (WBC) counts less than 1000/mm3, normal or mild elevated protein, and normal CSF/plasma sugar ratio. Three patients were found to have a virus in their CSF without pleocytosis. Virus isolations from CSF throat swabs and/or rectal swabs were performed in 65 patients, half of them (35/65, 53.8%) had positive results including echovirus type 9 (sixteen), echovirus type 30 (eighteen), and adenovirus type 3 (one). Echovirus type 9 was predominant during July and August whereas echovirus type 30 became predominant after September. All patients recovered spontaneously without any sequelae.

Purification of the microsomal Ca2(+)-ATPase from rat liver.

The Ca2(+)-ATPase from rat liver microsomes has been solubilized in Triton X-100 and purified to homogeneity by ficoll-sucrose treatment, column chromatography with agarose-hexane adenosine 5'-triphosphate Type 2, and high pressure liquid chromatography (HPLC). The purified enzyme obtained by this sequential procedure exhibited a 183-fold increase in specific activity. After ficoll-sucrose treatment, the activity of the Ca2(+)-ATPase was stable for at least two weeks when stored at -70 degrees C. In SDS-polyacrylamide gels, several fractions from HPLC chromatography showed a single band at a position corresponding to a molecular weight of about 107 kDa. This value is consistent with the molecular weight of the phosphoenzyme intermediate of endoplasmic reticulum (ER) Ca2(+)-ATPase. Further characterization of the ER Ca2(+)-ATPase was performed by western immunoblots. Antiserum raised against the 100-kDa sarcoplasmic reticulum (SR) Ca2(+)-ATPase cross-reacted with the purified Ca2(+)-ATPase from rat liver ER membranes.

Neuromyelitis optica (Devic's disease) report of one case.

Neuromyelitis optica also named Devic's disease is an acute combined optic neuritis and transverse myelitis. It is thought to be a variant of multiple sclerosis, but its clinical presentation probably has only one attack without further recurrence and exacerbation. We present a 12-year-old girl who suffered from sudden onset of lower extremeties weakness, sensory loss and blurred vision after a prodromal URI symptom. CSF examination showed mild pleocytosis, elevated immunoglobulins, mild elevation of protein concentration. No oligoclonal band was detected. Serum virology showed high titer of anti-EB virus antibody. Visual evoke potential showed prolong of latency and decreased amplitude of both eyes. After prednisolone treatment, her visual accuity began to improve on the 7th hospital day and motor function improved on the 11th hospital day. Two years later, she has normal visual accuity, normal motor function and shows no evidence of disease recurrence.

Nonspecific congenital myopathy (minimal change myopathy): a case report.

A 28-month-old male with generalized hypotonia and muscle weakness, a myopathic face, skeletal dysmorphism and delayed motor milestones from birth is reported. He gradually developed the ability of sitting and rolling over, but could not stand without support until 28 months. There was no intellectual impairment or seizures. Deep tendon reflexes were absent. The serum CK value, peripheral nerve conduction velocity and EMG were within normal limits. A muscle biopsy specimen showed mild variation in fiber size, and an increased number of type 2C fibers on histochemical examination, but no apparent abnormalities on electron microscopy. The baby was tentatively diagnosed as having minimal change myopathy or nonspecific congenital myopathy which is thought to be one of the congenital nonprogressive myopathies.