Immunoglobulin synthesis in vitro by cerebrospinal fluid cells in patients with meningoencephalitis of presumed viral origin.

Cerebrospinal fluid (CSF) cells from six patients with meningoencephalitis of presumed viral origin were incubated in the presence of labeled amino acids. The cells of two of the patients synthesized IgG, IgA, and IgM (one patient) in vitro. The CSF of these two patients had an elevated level of IgG with oligoclonal distribution. The newly synthesized IgG also had an oligoclonal distribution. CSF cells of the other four patients were not shown to synthesize immunoglobulins in vitro. The CSF of these patients had a normal level of IgG with polyclonal distribution. The results demonstrate that in some patients with virus meningoencephalitis an immunoglobulin synthesis takes place locally and that at least part of the IgG shows a restricted heterogeneity. The results also suggest the presence of stimulated lymphocytes in the CSF of the same patients.     

Germinal matrix hemorrhage of venous origin in preterm neonates

The rupture point of germinal matrix hemorrhage in premature neonates was examined by postmortem angiography. Thirteen cases of germinal matrix hemorrhage were injected via artery (four cases), vein (five cases), and both artery and vein simultaneously (four cases). Only material injected via vein leaked into the hemorrhage, which was confirmed by stereomicroscopic and histologic examination. This study suggests that germinal matrix hemorrhage is venous in origin.     

HIV-producing T cells in cerebrospinal fluid

In HIV-1-infected subjects, the magnitude of HIV-1 viral load in cerebrospinal fluid (CSF) correlates with the CSF white cell count. To determine whether HIV-1-producing T cells appear in CSF and whether their percentage and number correlate with viral load in CSF, we developed a flow cytometric assay that detects HIV-1-producing T cells by identifying intracellular p24 HIV-1 antigen. We found that most CSF T cells were not HIV-1 producing, even when cell-free viral load in CSF was high. Most activated T cells in CSF were also not HIV-1 producing, but the activated CD38+ CD4 T-cell fraction in CSF was independently associated with the fraction of HIV-1-producing T cells in CSF. We conclude that HIV-1-producing T cells appear in CSF and that their percentage and number correlate with cell-free viral load in CSF, even though the CSF total white cell count remains the best predictor for CSF viral load. In HIV-1 infection, CSF white cell counts seem to contain a large number of uninfected cells. White cell counts and viral load in CSF may result from systemic inflammation and immune activation.     

Possible origin of amniotic fluid constituents influencing fibrinolytic activity

An attempt was made to clarify the origin of the amniotic fluid constituents affecting fibrinolysis. For this purpose, saline extracts were prepared from various extra-fetal gestational tissues (EGT) as well as from various fetal excretory products (FEP) and their effect on an euglobinolytic system was studied in vitro. It was found that extracts from FEPs stimulated, whereas those from EGTs inhibited fibrinolysis. It is concluded that the fibrinolysis inhibiting amniotic fluid constituents originate from the placenta and the fetal membranes, whereas the fibrinolysis stimulating ones from various sources of fetal origin.     

Microvasculature in germinal matrix layer: its relationship to germinal matrix hemorrhage

Microvasculature in the germinal matrix layer (GML) was studied by postmortem microangiography and stereomicroscopic observation on cleared specimens in methyl salicylate. A number of arterioles from penetrating and medullary branches of main cerebral arteries were distributed in the GML, forming a fine network of arterioles anastomosing with each other. Many venules in the GML converged at a few points and drained into subependymal veins. The main converging zone of venules was situated inside the GML close to the border zone of the GML and adjacent cerebral parenchyma. The converging zone coincided with the site of GML hemorrhage. Thus, many factors, including both venous and arterial, appear to influence circulation in the GML; ultimately the converging zones become overloaded and rupture, leading to GML hemorrhage.     

Identification of Ehrlichia chaffeensis morulae in cerebrospinal fluid mononuclear cells.

We report a case of ehrlichiosis in a 72-year-old man who developed extreme lethargy, acute renal failure requiring hemodialysis, and respiratory insufficiency requiring intubation. Lumbar puncture performed on the second day of hospitalization revealed significant cellular pleocytosis. Ehrlichia morulae were tentatively identified in mononuclear cells in routinely processed Wright-stained cytospin preparations of cerebrospinal fluid (CSF). Identification was confirmed by a specific immunocytochemical staining procedure. Subsequent identification specifically as Ehrlichia chaffeensis morulae was established by polymerase chain reaction analysis, which revealed E. chaffeensis-specific DNA in CSF, bone marrow, and blood samples; by indirect fluorescent-antibody analysis, the patient developed an antibody titer of 32,768 against E. chaffeensis antigen. The patient responded to intravenous therapy with doxycycline and dexamethasone. Subsequently, neurologic, hematologic, renal, and pulmonary status had returned to baseline at follow-up 12 weeks after admission. To our knowledge, this is the first identification of E. chaffeensis morulae in CSF cells in an infected patient.     

Leukocyte survival in cerebrospinal fluid.

Delays in the laboratory examination of cerebrospinal fluid are commonly encountered in clinical medicine. The present studies were designed to evaluate changes in cerebrospinal fluid leukocyte counts relative to time elapsed before analysis. Neutrophil counts decreased most rapidly, being 68 +/- 10% (standard error of the mean) and 50 +/- 12% of initial values at 1 and 2 h, respectively. Lymphocyte and monocyte numbers were not significantly altered until 3 h.     

Assay of gentamicin in cerebrospinal fluid.

A comparison of standard curves obtained from a conventional plate diffusion assay method revealed significant differences when gentamicin standards were made up in different media. Standards made up in distilled water resulted in a curve which differed from that of standards made up in pooled human cerebrospinal fluid by a factor of up to 4. When the assay medium was supplemented with 0-5% sodium chloride, the difference between the two standard curves was reduced to a factor of about 1-5. The curve obtained from standards made up in 150 mM sodium chloride/4-5 mM calcium chloride correlated well with that from standards made up in cerebrospinal fluid. There was no evidence of gentamicin being bound to protein in the cerebrospinal fluid.     

Gelatinase activity of matrix metalloproteinases in the cerebrospinal fluid of various patient populations

We have studied the enzymatic gelatinolytic activity of matrix metalloproteinases (MMPs) present in cerebrospinal fluid (CSF) of samples obtained from 67 individuals, twenty-one nonneurological patients (considered controls) and 46 subjects with various neurological disorders e.g., vascular lesions, demyelination, inflammatory, degenerative and prion diseases. Biochemical characterization of MMPs, a family of neutral proteolytic enzymes involved in extracellular matrix modeling, included determination of substrate specificity and Ca+2 dependency, as well as the effects of protease inactivators, carboxylic and His (histidine) residue modifiers, and antibiotics. Whereas all CSF samples expressed MMP-2 (gelatinase A) activity, it corresponded in most cases (normal and pathological samples) to its latent form (proenzyme; pMMP-2). In general, inflammatory neurological diseases (especially meningitis and neurocisticercosis) were associated with the presence of a second enzyme, MMP-9 (or gelatinase B). Whereas MMP-9 was found in the CSF of every tropical spastic paraparesis patient studied, its presence in samples from individuals with vascular lesions was uncommon. Patients blood-brain barrier damage was ascertained by determining total CSF protein content using both, the conventional polyacrylamide gel electrophoresis procedure under denaturing conditions and capillary zone electrophoresis.     

Lymphoma cells in cerebrospinal fluid confirmed by chromosome analysis

Two cases of malignant lymphoma are reported, in which lymphoma cells were undergoing cell division in the cerebrospinal fluid. In each case it was possible to perform chromosome counts and karyotype analyses, and in this way to establish that a neoplastic clone was present.     

Blast-like cell compartment in carcinogen-induced proliferating bile ductules.

Small non-epithelial cells with morphological features of blast-like cells are found within a proliferating intrahepatic biliary system after institution in rats of a diethylnitrosamine, 2-acetylaminofluorene, partial hepatectomy carcinogenesis protocol. Two to three days after the partial hepatectomy step of the carcinogen protocol, the small blast-like cells are evident beneath a layer of bile ductule epithelial cells that line the walls of the bile ductules. The basally located small cells are not exposed to the bile ductule lumen or to the surrounding basal lamina. They ranged in size from 3.0 to 5.0 microns, exhibit an undifferentiated phenotype, including a high nucleus-to-cytoplasm ratio and no to minimal differentiated cytoplasmic and surface structures. Mitosis of blast-like cells are evident, and their nuclei express proliferating nuclear cell antigen. The ductal blast-like cells do not express cytokeratin 19, oval cell antigen 270.38, or actin immunoreactivity, in contrast to bile ductule epithelial cells. The basal cells, as well as bile ductule epithelial cells, are negative for a panel of T and B lymphocyte surface markers in contrast to lymphocytes present in the connective tissue stroma surrounding the bile ductules and throughout the hepatic parenchyma. Within some segments of the biliary system, some of the ductal blast-like cells increased in size to approximately 10 microns and showed increased amounts of cytoplasmic organelles and plasma membrane filapodia but did not develop the polarized phenotype of bile ductule epithelial cells (ie, apical microvilli, desmosomes, connections to bile ductule cells, and exposure to duct lumen); however, their nuclear morphology was essentially similar to the smaller basal cells. We also found bile ductules to contain two types of polarized epithelial cells, one with the characteristic oval nucleus of the oval/bile ductule epithelial cells and the other, transitional epithelial cells with a rounder nucleus and prominent nucleoli. The transitional cells exhibit a similar apical-basal polarity and antigenic phenotype as the oval/bile ductule epithelial cells. However, transitional cells are larger and have an overall less dense cytoplasm than the bile ductule epithelial/oval cells, and some show apical microvilli changes and small catalase-positive peroxisomes. These observations indicate that a greater diversity of cell types exist within intrahepatic bile ductules of rats treated with carcinogens. Furthermore, the nonpolarized ductal blast-like cells undergo proliferation and are significantly different in phenotype from other hepatic cells previously reported as candidates for liver progenitor cells.     

Fibrin-fibrinogen degradation products in cerebrospinal fluid.

An increase in low molecular weight fibrin-fibrinogen degradation products (FDP) was demonstrated in cerebrospinal fluid (CSF) from 17 of 18 patients with bacterial or viral meningitis compared with 29 patients without meningitis. The CSF also showed an increase in coagulation proteins of molecular weight less than 90000 (factors VII, IX, and plasminogen) but did not contain fibrinogen (MW 340000) or plasminogen activator. It is concluded that low molecular weight FDP in the CSF in infective meningitis result from leakage through a damaged blood-CSF barrier rather than from local digestion of fibrin deposited on the meninges.