Strain-specific restriction of the antiphagocytic property of group A streptococcal M proteins

Group A streptococcal M proteins are type-specific virulence factors that inhibit phagocytosis. We used two M proteins, M5 and Emm22, to analyze the influence of genetic background on the properties of M proteins. Mutant strains, engineered to lack these M proteins, were complemented with genes encoding the homologous or heterologous M protein, and the complemented strains were analyzed for phagocytosis resistance. Neither the M5 nor the Emm22 protein conferred phagocytosis resistance in the heterologous background, but they did do so in the homologous background. This was not due to lack of surface expression in the heterologous background. Moreover, the M5 and Emm22 proteins expressed in heterologous background appeared to have normal structure, since they were not affected in their ability to bind different human plasma proteins. In particular, M5 or Emm22 had normal ability to bind human complement inhibitors, a property that has been implicated in phagocytosis resistance. Results similar to those obtained with M5 and Emm22 were obtained in experiments with the M6 and Emm4 proteins. Together, these data suggest that the surface expression of M protein alone may not be sufficient to confer phagocytosis resistance and consequently that strain-specific factors other than M and Emm proteins may contribute to the ability of group A streptococci to resist phagocytosis.     

Molecular analysis in Japanese patients with Charcot-Marie-Tooth disease: DGGE analysis for PMP22, MPZ,and Cx32/GJB1 mutations

Charcot-Marie-Tooth disease (CMT) is a heterogeneous disorder and is traditionally classified into two major types, CMT type 1 (CMT1) and CMT type 2 (CMT2). Most CMT1 patients are associated with the duplication of 17p11.2-p12 (CMT1A duplication) and small numbers of patients have mutations of the peripheral myelin protein 22 (PMP22), myelin protein zero (MPZ), connexin 32 (Cx32/GJB1), and early growth response 2 (EGR2) genes. Some mutations of MPZ and Cx32 were also associated with the clinical CMT2 phenotype. We constructed denaturing gradient gel electrophoresis (DGGE) analysis as a screening method for PMP22, MPZ, and Cx32 mutations and studied 161 CMT patients without CMT1A duplication. We detected 27 mutations of three genes including 15 novel mutations; six of PMP22, three of MPZ, and six of Cx32. We finally identified 21 causative mutations in 22 unrelated patients and five polymorphic mutations. Eighteen of 22 patients carrying PMP22, MPZ, or Cx32 mutations presented with CMT1 and four of them with MPZ or Cx32 mutations presented with the CMT2 phenotype. DGGE analysis was sensitive for screening for those gene mutations, but causative gene mutation was not identified in many of the Japanese patients with CMT, especially with CMT1. Other candidate genes should be studied to elucidate the genetic basis of Japanese CMT patients.     

Differential effect on the sympathetic transmitter level in uterus and other organs of guinea-pig by drugs interfering with adrenergic nerve functions.

5 types of adrenergic blocking agents with different modes of action were administered and the rate of transmitter depletion was studied in different uterine regions of guinea-pig, and compared with that in the heart and submandibular gland by a combination of fluorescence histochemical and spectrofluorimetric methods. The tyrosine-hydroxylase inhibitor, H 44/68, as well as reserpine and guanethidine, produced a more efficient reduction in neuronal noradrenaline in the heart and submandibular gland than in the uterus. A differential action on the sympathetic transmitter, though less clearcut, was seen with 6-hydroxydopamine. The noradrenaline-depleting effect of metaraminol did not distinguish between those adrenergic nerves supplying the uterus from those of the control tissues. It is suggested that the differential effects may be related to a lower activity in the uterine adrenergic nerve system, probably because this system is to a large extent composed of short adrenergic neurons, which appear to have a particularly slow transmitter turnover.     

Primary malignant melanoma of the common bile duct

Biliary tract obstruction in a 30-year-old man was found to be caused by a malignant melanoma in the common bile duct. Melanin pigment was demonstrated by immunohistochemistry and electron microscopy. Extensive search for a primary malignant melanoma elsewhere was unsuccessful. No pigmented lesions had been removed previously. There were junctional changes in the mucosa of the common bile duct close to the tumor. The malignant melanoma in the common bile duct therefore is considered to be primary. Only one other case of primary malignant melanoma in the common bile duct has been described in the literature, whereas metastases to the major bile ducts in one autopsy study of malignant melanoma in the more common locations were found with a frequency of 6 per cent.     

The ventral spino-olivocerebellar system in the cat. V. Supraspinal control of spinal transmission

1. The transmission from the flexor reflex afferents (FRA) and from tracts running in the ipsilateral half of the spinal cord to the spino-olivocerebellar paths ascending through the ventral funiculus (VF-SOCPs; Oscarsson and Sj?lund, 1977) was compared with the transmission from these sources to segmental reflex arcs. The climbing fibre responses evoked in Purkinje cells by electrical stimulation of limb nerves and spinal tracts were monitored by recording the mass activity at the cerebellar surface simultaneously from several termination zones, while the activity in flexor motoneurones was recorded from a flexor nerve and the primary afferent depolarization from a dorsal filament. 2. Changes in the segmental reflex response were produced by release from the tonic inhibition of transmission from the FRA in decerebrate preparations and by conditioning electrical stimulation of dissected spinal funiculi containing inhibitory descending tracts. 3. The changes of the transmission from the FRA to two of the paths, the a- and b2-VF-SOCPs, parallelled the changes of the transmission to the segmental reflex arcs. On the other hand, the monosynaptic transmission from the FRA to the c1- and c3-VF-SOCPs was not significantly influenced by the inhibitory descending control systems. 4. The a- and b2-VF-SOCPs but not the c1- and c3-VF-SOCPs received polysynaptic excitation from tracts running in the ipsilateral half of the spinal cord. 5. The suggestion that the a- and b2-VF-SOCPs carry information related to interneuronal activity in segmental centres, whereas the c1- and c3-VF-SOCPs forward information mainly related to peripheral events (Andersson and Sj?lund, 1978) is supported by the present findings.     

The use of proteomics in biomarker discovery in neurodegenerative diseases

Biomarkers for neurodegenerative diseases should reflect the central pathogenic processes of the diseases. The field of clinical proteomics is especially well suited for discovery of biomarkers in cerebrospinal fluid (CSF), which reflects the proteins in the brain under healthy conditions as well as in several neurodegenerative diseases. Known proteins involved in the pathology of neurodegenerative diseases are, respectively, normal tau protein, beta-amyloid (1-42), synaptic proteins, amyloid precursor protein (APP), apolipoprotein E (apoE), which previously have been studied by protein immunoassays. The objective of this paper was to summarize results from proteomic studies of differential protein patterns in neurodegenerative diseases with focus on Alzheimer's disease (AD). Today, discrimination of AD from controls and from other neurological diseases has been improved by simultaneous analysis of both beta-amyloid (1-42), total-tau, and phosphorylated tau, where a combination of low levels of CSF-beta-amyloid 1-42 and high levels of CSF-tau and CSF-phospho-tau is associated with an AD diagnosis. Detection of new biomarkers will further strengthen diagnosis and provide useful information in drug trials. The combination of immunoassays and proteomic methods show that the CSF proteins express differential protein patterns in AD, FTD, and PD patients, which reflect divergent underlying pathophysiological mechanisms and neuropathological changes in these diseases.     

Radiographic quantification of alveolar bone level changes

The "random burst" theory has recently been proposed as an explanation of the pattern of periodontal disease progression. The theory predicts that the progression of bone loss at individual sites is not dependent upon previous bone loss and age. A longitudinal radiographic study was designed to test this hypothesis, and to describe the changes in bone level over 2 years in a group of 180 subjects (18-68 years of age) who were not under systematic periodontal treatment. The results indicated that 94% of the sites did not show significant changes in the alveolar bone level during the observation period. The mean annual bone loss for the total population was 0.11 mm. By regressing longitudinal bone loss upon age, it was shown that the rate of bone loss increased rapidly between 33 and 56 years of age while a different pattern was shown for the age intervals 18-32 and 57-68 years. Also, the rate of bone loss increased with increasing initial bone loss. This was less evident in the oldest age group. It was concluded that the progression of bone loss in the present material is consistent with a "burst" theory. However, the progression did not occur randomly with regard to previous loss of alveolar bone and time.     

Compatibility of parenteral nutrition solutions when mixed in a plastic bag

The administration of a single admixture of the components used in total parenteral nutrition has been considered more convenient and to give rise to less infection than the conventional regimen with separate infusions. The physico- chemical properties of several admixtures corresponding to commonly used regimens for complete parenteral nutrition have been studied. A 28 day toxicity study was carried out in the dog for one of these mixtures. It was concluded that Intralipid an be mixed with Vamin and dextrose solutions, supplemented with electrolytes, trace elements and vitamins provided that the admixtures are within certain defined ranges, prepared under strictly aseptic conditions and used within a limited time. Deviations from these ranges and component formulae should not be made without appropriate testing.     

CNS-induced natriuresis, neurohypophyseal peptides and renal dopamine and noradrenaline excretion in prehypertensive salt-sensitive Dahl rats

To identify defects in the salt-sensitive Dahl rat (Dahl-S), the natriuretic, catecholaminergic and pressor responses to 60-min elevation of the cerebroventricular sodium concentration (CNS-induced natriuresis) were compared between prehypertensive salt-sensitive Dahl-S and salt-resistant Dahl rats (Dahl-R). The plasma concentrations of the rat natriuretic hormone oxytocin, which has implications for the development of hypertension, and vasopressin (AVP) were also measured. Basal sodium and catecholamine excretion and mean arterial blood pressure (MAP) were similar in both strains. Sodium excretion during CNS stimulation increased more than 15-fold in Dahl-R but only 10-fold in Dahl-S. Dopamine excretion increased only transiently and similarly in both strains. Noradrenaline excretion and response to CNS stimulation were similar, suggesting a comparable sympathetic nervous activity between the strains. MAP increased comparably in Dahl-R and Dahl-S. Plasma AVP concentration was similar in both strains while plasma oxytocin concentration after CNS stimulation was more than 2-fold higher in Dahl-S than in Dahl-R. In conclusion, the prehypertensive Dahl-S has an attenuated natriuretic response to elevations of the cerebroventricular fluid sodium concentration and a higher plasma level of the natriuretic hormone oxytocin. Dopamine is not a mediator of CNS-induced natriuresis in neither strain. The attenuated natriuretic response may partly explain the salt-sensitivity in Dahl-S, and the higher plasma oxytocin value may either represent an effort to compensate for the deficient natriuretic response or reflect a primary defect in this system. Due to the known involvement of oxytocin in central MAP regulation in some hypertensive animal models, the findings warrant further investigation.