The molecular parameters of a meningococcal serogroup B protein-polysaccharide vaccine

Methods for the evaluation of the molecular parameters of B polysaccharide (B-PS) in meningococcal protein-polysaccharide vaccine of group B are proposed. The comparison of two proposed methods, the passive hemagglutination inhibition test and rocket immunoelectrophoresis (RIEP), has shown that the latter method has the highest degree of correlation with the chemical method of the detection of B-PS, which is often hindered by lactose added as a bulking agent. RIEP may be recommended for the standardization and control of the commercial preparations of group B meningococcal vaccine. B-PS contained in the vaccine is known to be in a highly polymeric state: its yield is 66.4 +/- 1.7% to Kd = 0.25 and 85.3 +/- 1.2% to Kd = 0.5. B-PS contained in the prepared vaccine has been found to be highly stable. Outer membrane proteins of meningococci, forming a noncovalent complex with capsular B-PS, seem to stabilize its structure and prevent the depolymerization of molecules.     

Increased expression of cyclin D1 and the Rb tumor suppressor gene in c-K-ras transformed rat enterocytes

Activating mutations in the c-K-ras gene occur in about 40% of human colorectal carcinomas, yet the role of this oncogene in tumorigenesis is not known. We have developed a model cell culture system to study this problem, utilizing the immortalized but non-tumorigenic epithelial cell line IEC18, originally derived from normal rat intestine epithelium. These cells were cotransfected with the drug resistance selectable marker tk-neo and the plasmid pMIKcys, which encodes a mini human c-K-ras gene (15 kb) containing a cysteine mutation at codon 12. Drug resistant clones were isolated. Clones which also expressed the activated c-K-ras gene displayed a transformed morphology, decreased doubling time, increased level of diacylglycerol, anchorage independent growth in soft agar and an aneuploid karyotype and they were also tumorigenic when injected into nude mice. These clones also displayed increased expression, at both the mRNA and protein levels, of cyclin D1 and Rb. These findings may be of clinical relevance since human colorectal tumors also frequently display increased expression of both cyclin D1 and Rb. This model system may be useful for understanding the role and interrelationship between activation of the c-K-ras oncogene and increased expression of cyclin D1 and Rb in colorectal tumorigenesis.     

Infinitary Logic and Inductive Definability over Finite Structures

The extensions of first-order logic with a least fixed point operator (FO + LFP) and with a partial fixed point operator (FO + PFP) are known to capture the complexity classes P and PSPACE respectively in the presence of an ordering relation over finite structures. Recently, Abiteboul and Vianu (in "Proceedings of the 23rd ACM Symposium on the Theory of Computing," 1991) investigated the relationship of these two logics in the absence of an ordering, using a machine model of generic computation. In particular, they showed that the two languages have equivalent expressive power if and only if P = PSPACE. These languages can also be seen as fragments of an infinitary logic where each formula has a bounded number of variables, L^ω_∞ω, (see, for instance, Kolaitis and Vardi, in "Proceedings of the 5th IEEE Symposium on Logic in Computer Science," pp. 156-167, 1990). We investigate this logic on finite structures and provide a normal form for it. We also present a treatment of Abiteboul and Vianu′s results from this point of view. In particular, we show that we can write a formula of FO + LFP that defines an ordering of the L^k_∞ω, types uniformly over all finite structures. One consequence of this is a generalization of the equivalence of FO + LFP and P from ordered structures to classes of structures where every element is definable. We also settle a conjecture mentioned by Abiteboul and Vianu by showing that FO + LFP is properly contained in the polynomial time computable fragment of L^ω_∞ω, raising the question of whether the latter fragment is a recursively enumerable class.     

Factors determining the long-term outcome of surgery for acromegaly

Seventy-nine patients with acromegaly were investigated before and after transsphenoidal adenomectomy, to determine the immediate and late outcome, the pre-operative features associated with a good result, and the accuracy of post-operative testing in predicting outcome. Pre-operative evaluation included basal growth hormone (GH), GH response to oral glucose tolerance test (OGTT), GH response to thyrotrophin-releasing hormone (TRH), tests of pituitary reserve, and pituitary scanning to assess tumour size. A few weeks after surgery, these tests were repeated. The patients were recalled for late assessment 1-13 years (median 86 months) after the operation. At the immediate postoperative testing, minimum GH after oral glucose was < or = 2 mU/l in 48.7%, < 5 mU/l in 76.3% and < 10 mU/l in 84.2%. Only 12 patients had GH > 10 mU/l. Basal GH was < or = 2 mU/l in 21%, < 5 in 59.2%, < 10 in 73.6% and < 20 in 90.8%. A minimum GH of < or = 2 mU/l during an OGTT was achieved in 67.4% of patients with intrasellar tumours, compared with 27.3% with extrasellar tumours. Basal GH and post-glucose GH correlated with the late outcome. GH response to TRH showed no correlation with outcome. IGF-1, which could not be assessed in detail, correlated with GH but was not a reliable indicator of outcome. Transsphenoidal adenomectomy is thus a very satisfactory treatment for acromegaly. Postoperative levels of basal growth hormone < 5 mU/l and post-glucose GH < or = 2 mU/l can be regarded as a biochemical cure. Postoperative radiotherapy is not required in patients who achieve a good result. The preoperative factors which significantly influenced the final outcome were basal GH, post-glucose minimum GH, tumour size and impaired pituitary reserve.     

Modulation of expression of glucose transporters GLUT3 and GLUT1 by potassium and N-methyl-D-aspartate in cultured cerebellar granule neurons

Depolarization is known to stimulate neuronal oxidative metabolism. As glucose is the primary fuel for oxidative metabolism in the brain, the entry of glucose into neural cells is a potential control point for any regulatory events in brain metabolism. Therefore, the effects of depolarizing stimuli, high K+ and N-methyl-D-aspartate (NMDA), were examined on the functional expression of glucose transporter isoforms GLUT1 and GLUT3 in primary cultured cerebellar granule neurons. Higher levels of glucose transport activity were observed in neurons cultured in 25 mM KCl (K25) compared to those in 5 and 15 mM KCl (K5 and K15). The elevated glucose transport activity correlated with increased levels of GLUT3 protein and, to a lesser extent, GLUT1. Both GLUT3 and GLUT1 were regulated at the level of mRNA expression. Addition of NMDA to K5 and K15 cultures increased both glucose uptake and GLUT3 protein levels, with smaller changes in GLUT1. NMDA effects were not additive with K25 effects. All these changes were observed only with chronic exposure of neurons to high K+ or NMDA; no acute effects on glucose uptake or transporter expression were found. Thus, chronic depolarization of primary cerebellar granule neurons acts as a stimulus for the expression of the neuronal GLUT3 glucose transporter isoform.     

Comparison of commercial kits for detection of cryptococcal antigen

Although kits to detect cryptococcal antigen are used widely to diagnose cryptococcal infection, the comparative performance of commercially available assays has not been evaluated in the past decade. Therefore, we compared the sensitives and specificities of five commercially available kits for detecting cryptococcal antigen (four latex agglutination test kits--Calas [Meridian Diagnostics])--Crypto-LA [International Biological Labs], Myco-Immune [MicroScan], and Immy [Immunomycologics]--and an enzyme immunoassay kit, Premier [Meridian Diagnostics]) with culture for the diagnosis of cryptococcal meningitis and fungemia. Of 182 cerebrospinal fluid (CSF) and 90 serum samples submitted for cryptococcal antigen and fungal culture, 49 (19 and 30 samples, respectively) from 20 patients had a culture positive for Cryptococcus neoformans. For CSF specimens, the sensitivities and specificities of all kits were comparable (sensitivity, 93 to 100%; specificity, 93 to 98%). There was a significant difference in sensitivities of the kits when serum samples were tested with the International Biological Labs and MicroScan kits, which do not pretreat serum with pronase. These kits were less sensitive (sensitivity, 83%) than the Immy and Meridian latex kits (sensitivity, 97%), which do pretreat with pronase. The sensitivity of the Meridian enzyme immunoassay kit was comparable to that of the pronase-containing latex kits. These kits were of equivalent specificities (93 to 100%) when testing serum. Some of the currently available kits have limitations that need to be recognized for proper interpretation of results. Specifically, the use of pronase on serum samples reduces the number of false-positive results, and a titer of < or = 1:4 can be a false-positive result when CSF samples are being tested.     

Determination of mean energies and impact parameters characteristic of charge-changing reactions

A guinea pig passage-adapted strain of the arena-virus Pichinde (adPIC) is highly virulent in inbred guinea pigs, whereas the related strain PIC3739 is attenuated. Both viruses were macrophage tropic and infected peritoneal, splenic, liver, and alveolar macrophages during experimental Pichinde virus infection. Infection with the virulent strain was associated with unlimited viral replication in the face of exaggerated delayed-type hypersensitivity response, manifested by the macrophage disappearance reaction. Histopathological lesions unique to adPIC-infected guinea pigs included intestinal villus blunting with mucosal infiltration by pyknotic debris-laden macrophages and apoptosis of crypt epithelial cells. Splenic red pulp necrosis was also significantly associated with adPIC infection but not PIC3739 infection. These findings may provide clues to the pathogenesis of a group of poorly understood human viral hemorrhagic fevers.     

Management of varicella infection during the course of inflammatory bowel disease

OBJECTIVE: To study the natural history and outcome of varicella infection developing in steroid treated inflammatory bowel disease. BACKGROUND: Varicella infection occurring in immunosuppressed or immunocompromised patients is a common problem with a significant mortality. Varicella infection during the course of inflammatory bowel disease has been reported in a small number of patients with at least one fatality. METHODS: Four young patients with inflammatory bowel disease who developed varicella infection while on immunosuppressive therapy, steroids, or azathioprine were studied. In each patient the infection was severe, and the three most recently treated patients received acyclovir. RESULTS: All four patients developed severe varicella infection while receiving immunosuppressive therapy for their disease. Three patients were treated with intravenous acyclovir with concomitant reduction of steroid dosage and recovered completely. One patient, treated in 1980 with antibiotics and reduction in steroids, did not receive acyclovir and also survived. CONCLUSIONS: Varicella infection is a relatively uncommon occurrence in inflammatory bowel disease. If varicella infection occurs, prompt diagnosis and treatment with acyclovir and concomitant reduction in immunosuppressive therapy (reduction in steroid dosage and discontinuation of azathioprine) should be initiated immediately to limit viremia and avoid fatal complications.     

Allogeneic bone marrow transplantation

Allogeneic bone marrow transplantation (BMT) after high-dose, marrow-ablative chemoradiotherapy has been established as the treatment of choice for various hematologic, neoplastic, and congenital disorders. The most common type of marrow graft is an allogeneic one from a sibling donor who has compatible human leukocyte antigen (HLA). Only 30% of patients requiring allogeneic BMT have an HLA-compatible sibling donor. Over the past few years, marrows from unrelated HLA-compatible donors have been used with increasing frequency and promising outcome in certain hematologic malignancies. Despite the morbidity and mortality associated with this treatment modality, allogeneic BMT may provide a 20% to 90% chance of long-term, disease-free survival to patients with a wide variety of neoplastic and abnormal marrow disorders.