Serum Matrix Metalloproteinase-2 Levels Indicate Blood–CSF Barrier Damage in Patients with Infectious Meningitis

Objective Protein components in cerebrospinal fluid (CSF) are maintained at a specific concentration by a dynamic gradient between the capillary and intrathecal spaces via the blood–cerebrospinal fluid barrier (BCB) in the brain and spinal cord. Permeability to proteins increases when there is structural damage to the BCB. Matrix metalloproteinase-2 (MMP-2; gelatinase A) has been shown to degrade type IV collagen, a major component of the cellular basement membrane. We analyzed α2 macroglobulin (α2M) indices and evaluated the relationship between α2M, as an indicator of BCB permeability, and MMP-2, which degrades the extra-cellular matrix in patients with infectious meningitis. Materials and Methods Albumin levels in CSF or serum were determined by turbidimetric immunoassay, or bromcresol green assay, respectively. α2M levels in CSF or serum were measured with enzyme-linked immunosorbent assay, or laser-nephelometry, respectively. Serum MMP-2 levels were determined by enzyme immuno assay. We calculated the α2M index, i.e. the ratio of α2M (CSF / serum) to albumin (CSF / serum; α2M in CSF / α2M in serum × albumin in serum / albumin in CSF). Results α2M indices were significantly increased in infectious meningitis compared to healthy controls (p < 0.05). They were highest in bacterial meningitis, and there was a significant difference between viral or mycotic and bacterial meningitis (p < 0.05). Serum MMP-2 levels were increased in infectious meningitis, being highest in bacterial meningitis, where they were significantly different from healthy controls (p < 0.05). There was a significant positive correlation between serum MMP-2 levels and α2M indices (r = 0.64, p < 0.0001). Conclusion Markedly increased levels of serum MMP-2 in infectious, especially bacterial, meningitis may reflect the degree of damage to the BCB.     

Evidence for a role of connexin 43 in trigeminal pain using RNA interference in vivo

The importance of glial cells in the generation and maintenance of neuropathic pain is becoming widely accepted. We examined the role of glial-specific gap junctions in nociception in the rat trigeminal ganglion in nerve-injured and -uninjured states. The connexin 43 (Cx43) gap-junction subunit was found to be confined to the satellite glial cells (SGCs) that tightly envelop primary sensory neurons in the trigeminal ganglion and we therefore used Cx43 RNA interference (RNAi) to alter gap-junction function in SGCs. Using behavioral evaluation, together with immunocytochemical and Western blot monitoring, we show that Cx43 increased in the trigeminal ganglion in rats with a chronic constriction injury (CCI) of the infraorbital nerve. Reducing Cx43 expression using RNAi in CCI rats reduced painlike behavior, whereas in non-CCI rats, reducing Cx43 expression increased painlike behavior. The degree of painlike behavior in CCI rats and intact, Cx43-silenced rats was similar. Our results support previous suggestions that increases in glial gap junctions after nerve injury increases nociceptive behavior but paradoxically the reduction of gap junctions in normal ganglia also increases nociceptive behavior, possibly a reflection of the multiple functions performed by glia.     

Utility of bedside artificial pancreas for postoperative glycemic control in cardiac surgery

Journal of Artificial Organs - Perioperative hyperglycemia, hypoglycemia, and high glycemic variability are independent risk factors for mortality in critically ill patients. After cardiac surgery,...     

Bovine herpesvirus 1 glycoprotein G is required for viral growth by cell-to-cell infection

The bovine herpesvirus 1 (BHV-1) US4 gene encodes glycoprotein G (gG), which is conserved in the majority of alphaherpesviruses. In order to identify the role of BHV-1 gG in the viral infection cycle, a gG minus BHV-1 mutant and its gG-positive revertant were constructed and their growth characteristics in Madin-Darby bovine kidney (MDBK) cells were compared. The gG minus mutant formed smaller plaques than the gG-positive BHV-1 in MDBK cells. When a monolayer culture of MDBK cells was infected with BHV-1 at a low multiplicity of infection and overlaid with semi-solid growth medium, under which adsorption of the mature virion released in the medium was inhibited, gG-positive BHV-1 multiplied, while the growth of the gG negative BHV-1 was severely inhibited. These data suggest that BHV-1 gG functions in direct cell-to-cell transmission mechanism of BHV-1 in tissue culture.     

A Complement Factor B Mutation in a Large Kindred with Atypical Hemolytic Uremic Syndrome

Purpose

Gain-of-function mutations in complement factor B (CFB) were recently identified in patients with atypical hemolytic uremic syndrome (aHUS), but are extremely rare. Our purpose is to describe a large kindred with aHUS associated with a CFB mutation and to further understand CFB-mutated aHUS patients.

Methods and Results

We report a large kindred in which 3 members had aHUS. This kindred revealed that 9 of 12 members, including 2 affected patients, had persistent activation of the alternative pathway with low complement component 3 and that those 9 members showed a CFB mutation (c.1050G?>?C, p.Lys350Asn) in exon 8. This missense mutation was heterozygous in 8 of them and homozygous in only one. From structural studies, this mutation is shown to be located in close proximity to the Mg²-binding site within a von Willebrand factor type A domain of CFB, resulting in a gain-of-function effect of CFB and predisposition to aHUS. At present, 2 of the 3 members with aHUS have maintained normal renal function for a long-term period.

Conclusions

This kindred illustrates that a CFB mutation (c.1050G?>?C, p.Lys350Asn) can result in aHUS. In the future, phenotype-genotype correlations and outcome in CFB-mutated aHUS patients need to be further investigated by accumulation of a number of cases.     

Two cases of partial trisomy 21 (pter-q22.1) without the major features of Down syndrome

We report two cases of partial trisomy 21 with clinical features distinct from Down syndrome (DS). These patients presented with moderate mental retardation and short stature, but the typical facial appearance of DS was not observed. Each patient had a similarly sized extra chromosome 21. We performed FISH analysis to examine whether deletions of reported approximately 5 Mb DS critical region (DSCR) might be associated with unusual clinical features in these cases. The results showed that each of their extra chromosomes 21 contained a distal part of chromosome 3p or 14q at the telomeric region of chromosome 21q. The translocation breakpoint of 21q for each patient was located on the centromeric side of DSCR (DSCR was deleted) and the sizes of partial trisomy 21 in respective patients are approximately 34.5 (21pter-q22.12) and approximately 33.0 Mb (21pter-q22.11). In one patient, the additional region of the short arm of chromosome 3 was 3pter-p26.1 from maternal origin, measuring approximately 9 Mb in size. The second patient had an extra 14q32.1-qter of maternal origin, measuring approximately 14 Mb in size. These are one of the shortest partial distal trisomy among reported cases. Taken together, two patients with partial trisomy 21 lack all of DSCR on 21q22, and their distinct clinical features are likely caused by the genes located at 21pter-q22.1 and the distal part of chromosome 3p or 14q.     

Revisited univariate delta check method for hematologic laboratories (I)--Usefulness for detection of specimen mix-up in patients with hematologic disorders

We evaluated the delta check method for hematologic laboratories to detect specimen mix-up. We selected 271 patients with hematologic disorders and two types of investigation were conducted. The first investigation comprised statistical analysis, while the second involved evaluation of the procedure. From parameters of white blood cell, red blood cell, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin(MCH), mean corpuscular hemoglobin concentration (MCHC), and platelets, MCV was considered to represent the best single marker to detect artificial mix-up. About 98% of MCV delta values from one patient were within 3 fl. Conversely, 40% of MCV delta values in artificial mixups exceed 10 fl. No correlations between time interval and MCV delta values were detected. However, some cases were observed in which MCV delta values changed markedly over a short period of time even though the samples originated from one patient. In conclusion, we recommend investigation for specimen mix-up in cases where MCV delta values exceed 4 fl.     

Polymorphisms and the differential antiviral activity of the chicken Mx gene

The nucleotide sequence of chicken Mx cDNA was reported earlier using the White Leghorn breed in Germany, but it showed no enhanced resistance to viruses. In this study, the nucleotide sequences of chicken Mx cDNA were determined in many breeds. A total of 25 nucleotide substitutions, of which 14 were deduced to cause amino acid exchanges, were detected, suggesting that the chicken Mx gene is very polymorphic. Transfected cell clones expressing chicken Mx mRNA were established after the Mx cDNA was constructed with an expression vector and introduced into mouse 3T3 cells, and the Mx genes from some breeds were demonstrated to confer positive antiviral responses to influenza virus and vesicular stomatitis virus. On the basis of the comparison among the antiviral activities associated with many Mx variations, a specific amino acid substitution at position 631 (Ser to Asn) was considered to determine the antivirally positive or negative Mx gene. Thus, a single amino acid substitution influences the antiviral activity of Mx in domesticated chickens.     

Involvement of an Skp-Like Protein,PGN_0300, in the Type IX Secretion System of Porphyromonas gingivalis

The oral Gram-negative anaerobic bacterium Porphyromonas gingivalis is an important pathogen involved in chronic periodontitis. Among its virulence factors, the major extracellular proteinases, Arg-gingipain and Lys-gingipain, are of interest given their abilities to degrade host proteins and process other virulence factors. Gingipains possess C-terminal domains (CTDs) and are translocated to the cell surface or into the extracellular milieu by the type IX secretion system (T9SS). Gingipains contribute to the colonial pigmentation of the bacterium on blood agar. In this study, Omp17, the PGN_0300 gene product, was found in the outer membrane fraction. A mutant lacking Omp17 did not show pigmentation on blood agar and showed reduced proteolytic activity of the gingipains. CTD-containing proteins were released from bacterial cells without cleavage of the CTDs in the omp17 mutant. Although synthesis of the anionic polysaccharide (A-LPS) was not affected in the omp17 mutant, the processing of and A-LPS modification of CTD-containing proteins was defective. PorU, a C-terminal signal peptidase that cleaves the CTDs of other CTD-containing proteins, was not detected in any membrane fraction of the omp17 mutant, suggesting that the defective maturation of CTD-containing proteins by impairment of Omp17 is partly due to loss of function of PorU. In the mouse subcutaneous infection experiment, the omp17 mutant was less virulent than the wild type. These results suggested that Omp17 is involved in P. gingivalis virulence.