Biomechanical Properties of Esophagus during Systemic Treatment with Epidermal Growth Factor in Rats

The epidermal growth factor (EGF) is a growth factor with effects on many cell types and tissue. Morphometric and passive biomechanical properties were studied in isolated segments of the esophagus in 22 EGF-treated rats and 12 control rats. The rats were divided into groups with EGF treatment for 2, 4, 7, and 14 days (n=6 for each group except n=4 for the 14 days EGF-treatment group) or saline treatment (n=3 for each group). The mechanical test was performed as a distension experiment in vitro where the whole esophagus was stretched to its in situ length and distended with pressures up to 10 cm H₂O using a ramp distension protocol. The pressure and outer diameter were recorded. Circumferential stress (force per area) and strain (deformation) were computed from the diameter and pressure data using the zero-stress state as reference. The zero-stress state was obtained by cutting esophageal rings radially. This caused the rings to open up into a sector. EGF induced pronounced morphometric changes, e.g., the wall thickness, wall cross-sectional area, and inner and outer circumferential lengths significantly increased during the EGF treatment. Histological analysis showed mucosa and submucosa growth during EGF treatment. The opening angle and residual strains increased with the highest value in the 14 days EGF-treated group (P < 0.05). The change in opening angle depended largely on the change in mucosa thickness. Furthermore, the circumferential stiffness of the esophagus reached a maximum after 7 days EGF treatment (P < 0.01). © 2003 Biomedical Engineering Society. PAC2003: 8719Rr, 8717Ee     

From rafts to crafts: membrane asymmetry in moving cells

Many important biological events, including the leukocyte-mediated immune response, wound repair, axon guidance and developmental patterning, involve persistent cell movement towards a directional signal, a process termed chemotaxis. Establishment of functional and spatial cell polarity is an absolute requirement for this response. We propose that redistribution of specific membrane microdomains, termed rafts, during cell migration is a pivotal step in achieving polarity. On the one hand, partitioning of molecules into rafts might help to localize proteins at the front or the rear of moving cells, and on the other hand, rafts might function as platforms for local activation and coordination of the signaling pathways involved in cell migration.     

Increased CD5-positive B lymphocytes in type I diabetes.

CD5+ B lymphocytes have been implicated in the production of polyspecific and monospecific antibodies that bind self-antigens, and increased proportions of this B cell subset occur in patients with some autoimmune diseases. We investigated the proportion of peripheral blood CD5+ B lymphocytes in type I diabetic patients. Compared with 18 age-matched healthy subjects, 11 out of 28 (39.2%) type I diabetic patients had increased proportions of circulating CD5. B lymphocytes with no alterations in the numbers of circulating B and T lymphocytes. Although all patients with increased CD5 B lymphocytes also had serum islet cell antibodies and/or insulin autoantibodies, the occurrence of increased proportions of CD5+ B lymphocytes and serum autoantibodies was not significantly correlated. Increased proportions of CD5+ B lymphocytes was not related to the time elapsed since the clinical onset of diabetes. In addition, regardless of being increased or normal, the proportion of CD5+ B lymphocytes appeared as a relatively constant phenotype after 1 year of follow-up studies at 3-month intervals in eight patients. Although the significance of these findings remains to be established, the possibility exists that CD5+ cells play a role in the pathogenesis of type I diabetes.     

Comparison of ViraPap, Southern hybridization, and polymerase chain reaction methods for human papillomavirus identification in an epidemiological investigation of cervical cancer.

In order to provide a reliable diagnosis for the presence and type of human papillomavirus (HPV) DNA in a case-control study of cervical cancer in Colombia and Spain, 926 cervical scrapes from female subjects were examined by ViraPap (VP) and Southern hybridization (SH), and 510 of these (263 cases and 247 controls) were also tested by polymerase chain reaction (PCR) using the HPV L1 consensus primers. HPV DNA prevalence was much higher in cases than in controls by each of the three tests. There was complete agreement between the results of the three tests for 64.9% of the 510 specimens; 53.5% were negative and 11.4% were positive (regardless of type) by all tests. An additional 29.0% of the specimens were positive by PCR: 19.4% by PCR alone, 6.7% by PCR and VP, and 2.9% by PCR and SH. SH and/or VP gave positive results for 6.0% of the specimens for which the PCR finding was negative: 2.7% by SH alone, 2.5% by VP alone, and 0.8% by both VP and SH. When specimens which were positive by VP alone or only by SH at low-stringency conditions were excluded, PCR confirmed all but four specimens which were positive by other tests. The concordance between type-specific diagnosis by SH and PCR was 86% when HPVs were typed in both tests. HPV-16 accounted for over 80% of the typed HPVs in each test. The presence of blood in case specimens did not appear to inhibit HPV positivity by VP or by PCR at the dilution tested. Low amounts of cellular DNA of specimens resulted in some underestimation of HPV positivity by VP and SH but not by PCR. Compared with that of PCR, the sensitivities for case specimens were 38% by SH and 50% by VP; the sensitivity for control specimens, although it could not be measured precisely because there were few positive specimens, appeared to be lower than for case specimens. It was concluded that PCR-based tests are best suited for epidemiological investigation of HPVs.     

Use of Computed Tomography Scans for Cochlear Implants

While 3-dimensional (3D) imaging by computed tomography has long been desirable for research and treatment of cochlear-implant patients, technical challenges have limited its wide application. Recent developments in scanner hardware and image processing techniques now allow image quality improvements that make clinical applications feasible. Validation experiments were performed to characterize a new methodology and its imaging performance.     

A TaqI polymorphism in the 3'UTR of the IL-12 p40 gene correlates with increased IL-12 secretion

Interleukin-12 (IL-12) is a key cytokine for the induction of Th1 immune responses. We evaluated whether a TaqI polymorphism in the 3'UTR of the IL-12 p40 gene affects secretion of IL-12 in vitro, and whether this polymorphism is associated with susceptibility to Crohn's disease (CD). IL-12 p40 and p70 secretion by monocytes in relation to genotype was determined in 63 healthy donors. Genotype and allele frequencies of the TaqI polymorphism in 150 CD patients were compared with 145 ethnically matched healthy controls (HC). No significant association was found between genotype and IL-12 p40 secretion after stimulation of monocytes with SAC+IFNgamma. In contrast, increasing IL-12 p70 secretion was found across the categories of non-carriers, heterozygotes and homozygotes for the variant allele (median values+/-SEM: 147+/-27, 282+/-51 and 482+/-34 pg/ml, respectively; P<0.005). The allele and genotype frequencies of this polymorphism in patients with CD did not differ statistically significantly from HC. The presence of a TaqI polymorphism in the IL12 p40 3'UTR correlates with increased in vitro IL-12 p70, but not p40 secretion. While this polymorphism does not appear to be correlated with susceptibility to CD in the limited population of patients tested here, it could influence the occurrence of the disease in certain subsets of patients.     

Predominant clonal evolution leads to a close parity between gene expression profiles and subspecific phylogeny in Trypanosoma cruzi

Inositol is an essential precursor for the formation of glycosyl-phosphatidylinositol (GPI)-anchors found in the majority of surface molecules in trypanosomatids, in addition to its requirement for phoshatidylinositol signal transduction pathways. In Leishmania donovani, high-affinity inositol transport is catalyzed by the active myo-inositol/H+ transporter MIT, which is driven by a proton gradient across the parasite membrane. We have characterized the substrate specificity and pharmacology of L. donovani MIT in vitro and in promastigote cultures. High substrate specificity of myo-inositol transport was shown in competition studies with 14 different monosaccharides and MIT function was unaffected by the structurally similar pentose sugars or hexoses. L-Fucose and D-xylose, both inhibitors of the Na+-dependent inositol transport system in the human host, did not affect MIT transport function in the parasite. Competition studies with eight different inositol isomers revealed that proton bonds between the C-2, C-3 and C-5 hydroxyl groups of myo-inositol and the transporter protein played a critical role for substrate recognition, and the C-3 hydroxyl oxygen appears to act as an electron donor to form an H-bond with a positive charge of the MIT permease. The cytotoxic inositol analogue 3-fluoro-myo-inositol was recognized by MIT with similar affinity as myo-inositol and showed an IC50 value of 42 +/- 8 microM in L. donovani cultures. Finally, substrate affinities of MIT revealed apparent Km values of 84 +/- 8 microM for myo-inositol and 5.4 +/- 0.9 nM for H+, equal pH 8.27 + 0.08, suggesting that the L. donovani myo-inositol/H+ symporter is fully activated at physiological pH in the sandfly midgut or macrophage phagolysosome. We conclude that Leishmania MIT constitutes an attractive target for delivery of cytotoxic inositol analogues and differs significantly from the sodium-coupled myo-inositol transport system of the human host.     

Cellular distribution of bovine leukemia virus proteins gp51SU, Pr72(env), and Pr66(gag-pro) in persistently infected cells

Monoclonal antibodies (mAbs) against bovine leukemia virus (BLV) mature proteins and precursors were used to map the localization of these proteins in persistently infected non-lymphocytic cell lines using immunofluorescence assay (IFA) and immuno-electron microscopy. IFA staining was observed in the basolateral surface of live FLK-BLV cells. When using a mAb against Pr66(gag-pro), mottled pinpoint fluorescence was seen in the cell surface of polarized cells, but no reaction was observed in cells undergoing mitosis. However, a mAb against Pr72(env) stained only mitotic cells and cellular fragments. Additionally, in these dividing cells, this envelope (Env) precursor polyprotein was not evenly distributed but concentrated predominantly in only one daughter cell. To the best of our knowledge, this observation has not been reported previously, either for BLV or for other retroviruses. The results of immunogold electron microscopy confirmed the specificity of the mAbs in the intracellular level. In infected cells, Pr72(env) and gp51SU were seen in proximity at the plasma membrane in incipient budding sites. Additionally, the mAb against Pr72(env) also reacted with Env precursor polyproteins in the mitochondria of BLV-bat(2) ultrathin sections. These mAbs may be used as a tool for mapping virus excretion sites in the cell surface of naturally or in vitro infected cells in the different stages of the cell cycle.     

Interaction between KIR3DL1 and HLA-B*57 supertype alleles influences the progression of HIV-1 infection in a Zambian population

KIR and HLA loci are both highly polymorphic, and some HLA class 1 products bind and trigger cell-surface receptors specified by KIR genes. We examined whether KIR genes act in concert with HLA-B locus to control HIV-1 infection in a sample of Zambian patients. DNA samples from 88 Zambian patients with HIV-1 were examined. Patients were classified as either slow progressors (SP; n = 54) or rapid progressors (RP; n = 34) to AIDS. All were typed for HLA-B and KIR genes. Our results reveal an association between B*57 supertype (B*57s, which includes B*57 and B*58 alleles) and delayed progression to AIDS (p = 0.0007 by pc = 0.015; OR = 5.25). We also observed an increase incidence of Bw4-I80 in patients with slow progression (p = 0.001 by pc = 0.003, OR = 5). This increase was found to be secondary to B*57s. The presence of both KIR3DL1 and B*57S has a significant effect on progression to AIDS (p = 0.0008; OR = 5.61). B*57s genotypes with another HLA-B allele different from those in the trans position, which also had a specificity different to Bw4-I80 (Bw4-T80 or Bw6), was also greater in the SP than in the RP group (p = 0.00003; OR = 10.11). The presence of the inhibitory allele KIR3DL1 in combination with the HLA-B*57s alleles that contain the Bw4-I80 epitope, has a highly protective effect against progression to AIDS in Zambian patients.     

Multiple macrodontic multituberculism

Ekman-Westborg and Julin [1974: Oral Surg 38:217-222], described multiple macrodontia and multituberculism affecting the teeth without other anomalies (E-WJ). We describe a Chilean case in a 12-year-old with the typical dental alterations and with histopathologic findings that include absence of predentin layer and prominent reduced enamel epithelium. E-WJ is not a syndrome and we propose "multiple macrodontic multituberculism" as a better name for this anomaly of uncertain etiology affecting only the crowns of the teeth.