Feral Swine Leptospira Seroprevalence Survey in Hawaii, USA, 2007-2009.

Leptospirosis is considered the most widespread of zoonotic diseases. It was a notifiable disease in the United States until 1995 and was reinstated to the list of nationally notifiable diseases in 2014. During the time of national surveillance, Hawaii consistently led the nation in reported annual incidence rates. Leptospirosis has remained a reportable disease in Hawaii. Significant changes have been documented since the early 1970s in the predominant serogroup infecting humans in Hawaii: infections due to Icterohaemorrhagiae have declined while infections due to Australis have increased. A recent study from Hawaii demonstrated that Australis was an uncommon infecting serogroup for small mammal hosts. Swine have not been previously studied in Hawaii but are well-recognized maintenance hosts for leptospires belonging to the Australis serogroup. This study was undertaken to assess the prevalence of Leptospira antibody in feral swine in Hawaii. From January 2007 through December 2009, blood samples were collected opportunistically from feral swine. Using the microscopic agglutination test, we found antibody titres ≥1 : 100 to leptospires in 272 (33.8%) of 804 feral swine. The most frequently reacting serovars to the swine sera were Icterohaemorrhagiae (Icterohaemorrhagiae serogroup) (41.5%) and Bratislava (Australis serogroup) (33.8%). The high seroprevalence and presumptively infecting serovars suggest a link between swine and human infection.

Cancer-associated retinopathy.

Patients with systemic cancer may have a variety of ocular complaints. Most commonly these are metastases or adverse effects of therapy. Paraneoplastic syndromes, like cancer-associated retinopathy, rarely cause ophthalmic symptoms. We describe a patient with a malignant mixed mullerian tumor and cancer-associated retinopathy who had circulating serum antibodies to recoverin and cells positive for recoverin in the tumor. We discuss the typical clinical symptoms as well as the pathophysiology of this uncommon disorder.

Atypical central nervous system lymphoma at the cranial base: report of four cases.

OBJECTIVE AND IMPORTANCE: Primary central nervous system lymphoma is a disease with increasing incidence. Atypical presentations are becoming more frequent. CLINICAL PRESENTATION: Three patients bearing cavernous sinus lesions presented initially with periorbital pain and diplopia. Tolosa-Hunt syndrome was the initial presumptive diagnosis for two patients, and meningioma was the diagnosis for the third patient. A fourth patient presented with left ear pain, and a mass in the left internal auditory canal was thought to represent an acoustic neuroma. INTERVENTION: For all four patients, an operative pathological diagnosis was obtained and was compatible with central nervous system lymphoma. All patients were treated with osmotic blood-brain barrier disruption with intra-arterial chemotherapy using a methotrexate-based regimen. CONCLUSION: All four cases included atypical presentations of lymphoma. These cases again illustrate that a correct diagnosis cannot be obtained based only on imaging and clinical findings.

Effects of Gd-DTPA after osmotic BBB disruption in a rodent model: toxicity and MR findings.

OBJECTIVE: This experiment was done to evaluate the gross neurotoxicity of intravenous Gd-DTPA administered in conjunction with osmotic blood-brain barrier (BBB) disruption and to image a human small cell lung carcinoma intracerebral tumor xenograft before and after osmotic BBB disruption. MATERIALS AND METHODS: Neurotoxicity studies were performed in normal Sprague-Dawley rats following osmotic BBB disruption by the injection of 25% mannitol in the right internal carotid artery and intravenous administration of Gd-DTPA (n = 10). Animals were observed for major neurologic changes such as seizure or substantial motor defects, and after death neuropathologic examination was performed. Human small cell lung carcinoma cells were implanted intracerebrally in athymic nude rats (n = 4). Gadopentetate dimeglumine was injected intravenously and serial T1-weighted images were obtained. Blood-brain barrier disruption was produced in each animal, followed by a second dose of intravenous Gd-DTPA, and imaging studies were repeated. RESULTS: No gross neurologic toxicity was observed. Tumors showed dense enhancement in a small area, and BBB disruption resulted in marked enhancement in most of the gray matter of the right cerebral hemisphere. CONCLUSION: Gadopentetate dimeglumine appears to be safe in doses up to 21 mmol/m2 in conjunction with barrier disruption in rats. A human small cell lung carcinoma intracerebral xenograft provides a useful method to study brain tumors.

Advances in the treatment of superficial candida infections.

This article reviews recent controlled studies of treatment for candida infections of the skin and mucous membranes. In general, excellent agents are available for treatment, with reasonable alternatives for the uncommon emergence of resistant strains. Cutaneous infections in immunocompetent patients are effectively treated with topical agents. The major clinical problems that remain include frequently recurrent vulvovaginal candidiasis, and oroesophageal candida infections in patients with malignancies and immunosuppression. These often require systemic therapy. A number of treatment strategies have been investigated and are reviewed.

Distinct infrared spectroscopic patterns of human basal cell carcinoma of the skin.

Infrared spectroscopy combined with high pressure (pressure-tuning infrared spectroscopy) was applied to the study of paired sections of basal cell carcinomas (BCC) and normal skin from ten patients. Atmospheric pressure IR spectra from BCC were dramatically different from those from the corresponding normal skin. Compared to their normal controls, BCCs displayed increased hydrogen bonding of the phosphodiester group of nucleic acids, decreased hydrogen bonding of the C--OH groups of proteins, increased intensity of the band at 972 cm-1, a decreased intensity ratio between the CH3 stretching and CH2 stretching bands, and accumulation of unidentified carbohydrates. Some of these changes are shared by all human epithelial malignancies studied to date, while some others appear as yet unique to basal cell carcinoma. The diagnostic value of infrared spectroscopy in BCC remains to be determined.

Human skin mast cell carboxypeptidase: functional characterization, cDNA cloning, and genealogy.

We functionally characterized human skin mast cell carboxypeptidase A (MC-CPA), and explored its evolutionary relationship to other carboxypeptidases to understand further the structural basis for the substrate preferences of this enzyme. Purified human skin MC-CPA displayed more activity than did bovine pancreatic carboxypeptidase A (CPA) against carboxyl-terminal leucine residues, about equal activity with phenylalanine and tyrosine residues, and no activity with tryptophan or alanine. To correlate kinetic data with structure, we isolated and sequenced a cDNA encoding MC-CPA from human skin, and directly sequenced 30% of the purified protein. These sequences agreed with that of human lung MC-CPA, and further support the evidence for a single MC-CPA gene in humans. Four amino acid replacements, resulting in a net positive change in non-hydrogen atoms in the S1' subsite of MC-CPA, were associated with less alteration in substrate specificity, relative to bovine CPA, than might be expected from studies using rat CPA1 and CPA2. We noted two consensus N-linked glycosylation sites in human MC-CPA that are not found in rat and mouse MC-CPA, or in bovine CPA; that at least one of these sites is glycosylated in vivo was verified by N-glycosidase F treatment, lentil lectin binding, and Concanavalin A-Sepharose chromatography. Evolutionary trees constructed from the known carboxypeptidase sequences suggested that MC-CPA most likely evolved from a carboxypeptidase B-like enzyme, independent of the pancreatic CPA. Thus, in the carboxypeptidase gene family, MC-CPA displays a unique genealogy and several amino acid replacements in its S1' binding pocket that result in substrate specificity quite similar to bovine CPA.

MR of primary CNS lymphoma in immunologically normal patients.

PURPOSE: To describe the MR findings of primary CNS lymphoma. METHODS: MR scans of 20 patients with histologically proved primary CNS lymphoma were reviewed. We evaluated the size, multiplicity, signal intensities, and enhancement characteristics of the lesions. We divided the lesions into an enhancing area referred to as Zone 1 and abnormal signal surrounding this, referred to as Zone 2. RESULTS: Primary CNS lymphoma presented as solitary enhancing lesions in 40% of the patients and multiple lesions in 40%. Thirty-three separate lesions were visible: 58% abutted the ventricular system, 76% showed a homogenous enhancement pattern, and 79% showed marked enhancement. In 64% of the lesions, Zone 1 and Zone 2 showed different signal intensities on T1-weighted images. CONCLUSIONS: Primary CNS lymphoma usually presents as solitary or multiple dense homogenous enhancing lesions that abut an ependymal surface. These lesions can be divided into an enhancing area and an area of surrounding abnormal signal. These two areas often have different signal intensities on unenhanced T2-weighted images. These findings are sufficiently suggestive of the diagnosis of primary CNS lymphoma that a needle biopsy be performed based on these findings and appropriate therapy can then be instituted.

Protease composition of exocytosed human skin mast cell protease-proteoglycan complexes. Tryptase resides in a complex distinct from chymase and carboxypeptidase.

We characterized the release and the protease composition of high m.w. complexes released from dispersed human skin mast cells, under conditions that did not disrupt the binding of proteases to proteoglycan. The net percent release ratio of tryptase to histamine, after anti-IgE and calcium ionophore A23187 stimulation was higher than those for chymase or carboxypeptidase. This was explained by the greater cell association of carboxypeptidase and chymase, compared with tryptase, after mast cell degranulation and/or differential cosedimentation of the proteases with mast cells, because treatment of activated mast cells with 1 M NaCl increased the release ratios of chymase and carboxypeptidase more than that of tryptase. Tryptase, after release, was stable in 0.12 M NaCl and had a molecular mass of approximately 200 to 250 kDa, suggesting that it was bound to proteoglycan. We demonstrated that complexes containing chymase and carboxypeptidase were separable from tryptase-containing complexes by gel filtration and by affinity chromatography. First, on fast protein liquid chromatography, released tryptase filtered at a molecular mass of approximately 200 to 250 kDa, compared with chymase and carboxypeptidase at 400 to 560 kDa. Second, by using affinity chromatography with immobilized antitryptase mAb in 0.15 M NaCl, carboxypeptidase and chymase activities were recovered primarily in the effluent and washes of an antitryptase antibody affinity column and cofiltered at 400 to 560 kDa. Tryptase was recovered only in the eluate. Finally, by using potato tuber carboxypeptidase inhibitor-Sepharose affinity chromatography, tryptase activity was found primarily in the effluent and washes, filtered at a molecular mass of 200 kDa on fast protein liquid chromatography, and was stable in 0.12 M NaCl buffer at 37 degrees C. Carboxypeptidase and chymase activities were found primarily in the eluate. These findings suggest that tryptase and carboxypeptidase/chymase reside in distinct macromolecular complexes. Separate complexes containing these proteases may help explain previous ultrastructural observations in which the distributions of chymase and tryptase within a single granule did not always coincide.

Isolation of a neuropeptide-degrading carboxypeptidase from the human stomach.

The aim of this investigation was to isolate and characterize a neuropeptide-degrading carboxypeptidase from the muscular and mucosal layers of the human stomach. The carboxypeptidase was solubilized from membrane preparations of gastric muscle and mucosa using Triton X-100. The detergent-solubilized enzyme was purified to apparent electrophoretic homogeneity by affinity chromatography using lisinopril or potato carboxypeptidase inhibitor as an affinity ligand. The enzyme had an apparent molecular weight of 34,300 and was bound by concanavalin A and is thus a glycoprotein. The carboxypeptidase removed C-terminal leucine, phenylalanine, or tryosine residues from peptides including angiotensin I, [Leu5]enkephalin, kinetensin, neuromedin N, neurotensin, and xenopsin. It had an alkaline pH optimum and was inhibited by lisinopril, potato carboxypeptidase inhibitor, ethylenediaminetetraacetic acid, 1,10-phenanthroline, and 8-hydroxyquinoline. Immunoblotting indicated that the gastric carboxypeptidase cross-reacted with an antibody raised against a carboxypeptidase isolated from mast cells of human skin. The gastric carboxypeptidase released from gastric mast cells upon degranulation may act to degrade and inactivate neuropeptides in the stomach wall.

Effects of gadopentetate dimeglumine administration after osmotic blood-brain barrier disruption: toxicity and MR imaging findings.

Osmotic blood-brain barrier disruption with intraarterial chemotherapy has been shown to be beneficial in the treatment of malignant brain tumors. Imaging blood-brain barrier disruption is necessary to document the extent and degree of disruption and to correlate disruption with drug delivery. The present study evaluated blood-brain barrier disruption with gadopentetate dimeglumine-enhanced MR imaging and the associated toxicity of gadopentetate dimeglumine administration. Blood-brain barrier disruption was performed in seven dogs for imaging analysis and 17 dogs for toxicity evaluation. In the absence of gadopentetate dimeglumine administration, blood-brain barrier disruption could not be imaged. Enhanced MR imaging with a gadopentetate dimeglumine dose of 0.1 mmol/kg provided good images of disruption at an imaging time of 3 hr after disruption. However, when gadopentetate dimeglumine was given intravenously in conjunction with osmotic blood-brain barrier disruption, there was a statistically significant (p = .02) dose-dependent increase in the frequency of seizures, with 50% of the animals who received 0.1 mmol/kg and 75% who received 0.2 mmol/kg developing delayed seizures. Our findings show that, as with ionized iodinated CT contrast agents, gadopentetate dimeglumine is associated with toxicity when used in conjunction with osmotic blood-brain barrier disruption in dogs. Such toxicity may be a contraindication to the use of gadopentetate dimeglumine for monitoring patients with osmotically induced disruption of the blood-brain barrier.

Human mast cell proteases hydrolyze neurotensin, kinetensin and Leu5-enkephalin.

Purified mast cell carboxypeptidase cleaved the C-terminal leucines from Leu5-enkephalin (Leu-ENK), neurotensin (NT), and kinetensin (KT), with Km values of 36, 16, and 15 microM, and kcat values of 44, 51, and 53 s-1, respectively. To better predict potential in vivo hydrolysis products generated by mast cell proteases, these peptides were incubated with released skin mast cell supernatants. Leu5-enkephalin was hydrolyzed only by carboxypeptidase. Kinetensin was cleaved by tryptase, chymase, and carboxypeptidase to yield KT(1-3), KT(1-7), KT(1-8), KT(4-7), and KT(4-8), the last two peptides by the concerted action of two of the proteases. NT(1-11) and NT(1-12) were generated from neurotensin by chymase and carboxypeptidase, respectively.

Human mast cell carboxypeptidase. Selective localization to MCTC cells.

Two murine mAb were prepared against human mast cell carboxypeptidase (HMC-CP) purified from human skin, and were termed CP1 and CP2, respectively. Double immunohistochemical labeling of Carnoy's-fixed sections of human skin, lung, and gastrointestinal tissue with CP1 and CP2, respectively, and with a murine monoclonal antitryptase antibody demonstrated that HMC-CP was selectively present in a subset of human mast cells. Double labeling experiments with CP1 and CP2, respectively, and a murine anti-chymase mAb demonstrated the presence of HMC-CP in the tryptase-positive, chymase-positive mast cell type (MCTC) only. Immunohistochemical labeling of peripheral blood leukocytes resulted in staining of monocytes with CP2 but not with CP1. In addition to chymase and a cathepsin-G like proteinase, HMC-CP is another neutral protease that is selectively present in the MCTC tryptase-positive, chymase-positive mast cells type of mast cell, thus extending the biochemical definition of human mast cell heterogeneity.

Human mast cell tryptase: multiple cDNAs and genes reveal a multigene serine protease family.

Three different cDNAs and a gene encoding human skin mast cell tryptase have been cloned and sequenced in their entirety. The deduced amino acid sequences reveal a 30-amino acid prepropeptide followed by a 245-amino acid catalytic domain. The C-terminal undecapeptide of the human preprosequence is identical in dog tryptase and appears to be part of a prosequence unique among serine proteases. The differences among the three human tryptase catalytic domains include the loss of a consensus N-glycosylation site in one cDNA, which may explain some of the heterogeneity in size and susceptibility to deglycosylation seen in tryptase preparations. All three tryptase cDNAs are distinct from a recently reported cDNA obtained from a human lung mast cell library. A skin tryptase cDNA was used to isolate a human tryptase gene, the exons of which match one of the skin-derived cDNAs. The organization of the approximately 1.8-kilobase-pair tryptase gene is unique and is not closely related to that of any other mast cell or leukocyte serine protease. The 5' regulatory regions of the gene share features with those of other serine proteases, including mast cell chymase, but are unusual in being separated from the protein-coding sequence by an intron. High-stringency hybridization of a human genomic DNA blot with a fragment of the tryptase gene confirms the presence of multiple tryptase genes. These findings provide genetic evidence that human mast cell tryptases are the products of a multigene family.

A new sign of neurofibromatosis on magnetic resonance imaging of children.

Ten pediatric patients with clinically proved neurofibromatosis underwent magnetic resonance imaging of the brain. Seven of these patients had lesions of increased signal intensity on T2-weighted images in the globus pallidus, brain stem, or cerebellar white matter. The lesions did not correlate with results of the neurologic examination or with developmental status. These abnormalities most likely represent hamartomas and should be recognized as part of the diagnostic spectrum of neurofibromatosis.

Human mast cell carboxypeptidase. Purification and characterization.

A carboxypeptidase activity was recently identified in highly purified human lung mast cells and dispersed mast cells from skin. Using affinity chromatography with potato-tuber carboxypeptidase inhibitor as ligand, mast cell carboxypeptidase was purified to homogeneity from whole skin extracts. The purified enzyme yielded a single staining band of approximately 34,500 D on SDS-PAGE. Carboxypeptidase enzyme content estimated by determination of specific activity, was 0.5, 5, and 16 micrograms/10(6) mast cells from neonatal foreskin, adult facial skin, and adult foreskin, respectively. Human mast cell carboxypeptidase resembled bovine carboxypeptidase A with respect to hydrolysis of synthetic dipeptides and angiotensin I, but was distinguished from carboxypeptidase A in its inability to hydrolyze des-Arg9 bradykinin. The amino acid composition of human mast cell carboxypeptidase was similar to the composition of rat mast cell carboxypeptidase. The amino-terminal amino acid sequence of mast cell carboxypeptidase demonstrated 65% positional identity with human pancreatic carboxypeptidase B, but only 19% with human carboxypeptidase A. Thus, human mast cell carboxypeptidase is a novel member of the protein family of zinc-containing carboxypeptidases, in that it is functionally similar but not identical to bovine carboxypeptidase A, but has structural similarity to bovine and human pancreatic carboxypeptidase B.