Epitope mapping and biological function analysis of antibodies produced by immunization of mice with an inactivated Chinese isolate of severe acute respiratory syndrome-associated coronavirus (SARS-CoV)

Inactivated severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been tested as a candidate vaccine against the re-emergence of SARS. In order to understand the efficacy and safety of this approach, it is important to know the antibody specificities generated with inactivated SARS-CoV. In the current study, a panel of twelve monoclonal antibodies (mAbs) was established by immunizing Balb/c mice with the inactivated BJ01 strain of SARS-CoV isolated from the lung tissue of a SARS-infected Chinese patient. These mAbs could recognize SARS-CoV-infected cells by immunofluorescence analysis (IFA). Seven of them were mapped to the specific segments of recombinant spike (S) protein: six on S1 subunit (aa 12-798) and one on S2 subunit (aa 797-1192). High neutralizing titers against SARS-CoV were detected with two mAbs (1A5 and 2C5) targeting at a subdomain of S protein (aa 310-535), consistent with the previous report that this segment of S protein contains the major neutralizing domain. Some of these S-specific mAbs were able to recognize cleaved products of S protein in SARS-CoV-infected Vero E6 cells. None of the remaining five mAbs could recognize either of the recombinant S, N, M, or E antigens by ELISA. This study demonstrated that the inactivated SARS-CoV was able to preserve the immunogenicity of S protein including its major neutralizing domain. The relative ease with which these mAbs were generated against SARS-CoV virions further supports that subunit vaccination with S constructs may also be able to protect animals and perhaps humans. It is somewhat unexpected that no N-specific mAbs were identified albeit anti-N IgG was easily identified in SARS-CoV-infected patients. The availability of this panel of mAbs also provided potentially useful agents with applications in therapy, diagnosis, and basic research of SARS-CoV.     

Conservative management of early endometrial adenocarcinoma with repeat curettage and hormone therapy under assistance of hysteroscopy and laparoscopy

We report a rare case of early-stage endometrial adenocarcinoma in a 22 year old nullipara with polycystic ovaries undergoing conservative treatment. Pretreatment evaluation including tumour grade, depth of myometrial invasion, tumour size, hormone receptor status and flow cytometric analysis indicated a favourable prognosis. The patient underwent repeat endometrial curettage and a 6 month period of therapy with megestrol acetate and tamoxifen. A combination contraceptive pill was then prescribed to ensure withdrawal of the menstrual cycle thereafter. Now, 1 year after the last curettage, there is no evidence of disease. During the treatment period, hysteroscopy allowed for a more precise approach in panoramically examining the tumour nest in the endometrial cavity, and the subsequent endometrial response to hormone therapy. Laparoscopy using bulldog clamps applied to the isthmic portion of the Fallopian tubes prevented i.p. spread of endometrial tissue from retrograde regurgitation during hysteroscopy. Laparoscopic ovarian electrocautery resulted in the reduction of abnormal hypervascularization on the surface of polycystic ovaries postoperatively but caused a peri-ovarian adhesion complication. It is interesting that this case posed a unique opportunity to demonstrate the tumour regression under the assistance of laparoscopy and hysteroscopy.      

Evidence for object permanence in the smooth-pursuit eye movements of monkeys

We recorded the smooth-pursuit eye movements of monkeys in response to targets that were extinguished (blinked) for 200 ms in mid-trajectory. Eye velocity declined considerably during the target blinks, even when the blinks were completely predictable in time and space. Eye velocity declined whether blinks were presented during steady-state pursuit of a constant-velocity target, during initiation of pursuit before target velocity was reached, or during eye accelerations induced by a change in target velocity. When a physical occluder covered the trajectory of the target during blinks, creating the impression that the target moved behind it, the decline in eye velocity was reduced or abolished. If the target was occluded once the eye had reached target velocity, pursuit was only slightly poorer than normal, uninterrupted pursuit. In contrast, if the target was occluded during the initiation of pursuit, while the eye was accelerating toward target velocity, pursuit during occlusion was very different from normal pursuit. Eye velocity remained relatively stable during target occlusion, showing much less acceleration than normal pursuit and much less of a decline than was produced by a target blink. Anticipatory or predictive eye acceleration was typically observed just prior to the reappearance of the target. Computer simulations show that these results are best understood by assuming that a mechanism of eye-velocity memory remains engaged during target occlusion but is disengaged during target blinks.     

Heating patterns of microwave applicators in inhomogeneous arm and thigh phantoms.

Heating rate (HR) patterns in cylindrical structures were studied with inhomogeneous limb phantoms. These phantoms, arm and thigh models consisting of fat, bone, and muscle material, were heated with Clini-Therm L, M, and MS applicators at 915 MHz. The thigh model is 18 cm in diameter with 2.65-cm-thick fat on the outside and a 4-cm-diam bone in the center. The arm model is 9 cm in diameter with 1.35-cm-thick fat and 2-cm-diam bone. All models are 29 cm long with phantom muscle in the space between fat and bone and were heated with their long axes parallel or perpendicular to the E field. HR patterns in the transverse and longitudinal planes were obtained thermographically. A large water bolus, with the water channels parallel to the E field, was used in every case. In the thigh model, maximum heating was in the muscle for both L and M applicators when the E field was parallel to the long axis. When it was perpendicular, the maximum heating occurred in the fat layer. However, the peak HR in the fat remained about the same for both E field orientations. For the small applicator, heating was mostly in the fat, and the two field orientations did not cause much difference in the heating pattern. In the arm model, the maximum heating of all three applicators occurred mostly in the muscle for both E-field orientations. However, the maximum HR was reduced by a factor of 2 to 3 when the E field was perpendicular rather than parallel to the long axis.(ABSTRACT TRUNCATED AT 250 WORDS)     

The significance of small erythrocytes.

Small erythrocytes (mean corpuscular volume less than 80 mu-3 by the Coulter Model S) were found in 222 (2.75%) of 8,086 consecutive patients admitted to a large suburban general hospital. Forty-five (20.3%) of these 222 patients had laboratory findings consistent with thalassemia. Seventy-six (31.2%) were found to be iron deficient. Patients whose hemoglobin values were below 9.0 Gm. per 100 ml. were more likely to be iron deficient. The hemoglobin A2 values were significantly lower in iron-deficient than in non-iron-deficient patients. Although the mean corpuscular volume is much lower and the erythrocyte count is higher in thalassemia than in iron deficiency, hematologic values obtainable from the Counlter S (such as MCV, erythrocyte count, and hemoglobin) alone are not valuable in differentiating thalassemia from iron deficiency.     

Genetic changes in neoplasms arising in congenital melanocytic nevi: differences between nodular proliferations and melanomas

Large congenital melanocytic nevi (CMN) are at an increased risk of developing melanoma. Several forms of secondary proliferations can arise in congenital nevi on rare occasions. Although some of these closely resemble melanoma both clinically and histologically, metastasis is rare. We used comparative genomic hybridization to analyze chromosomal aberrations in different types of proliferations arising in CMN and compared them to typical congenital nevi, clear-cut melanomas arising in congenital nevi, as well as primary cutaneous melanomas that were not associated with a CMN. Cases of CMN and CMN with secondary proliferations were assigned to six groups according to the predominant histological pattern: group I, bland congenital nevi (n = 6); group II, congenital nevi with foci of increased cellularity (n = 4); group III, CMN with a proliferation simulating superficial spreading melanoma in situ (n = 3); group IV, CMN with a proliferation simulating nodular melanoma (n = 9); group V, proliferating neurocristic hamartoma (n = 1); and group VI, melanoma arising in congenital nevus (n = 6). No aberrations were found in groups I to III, whereas seven of nine cases of group IV, and one of one case of group V, showed aberrations. In group IV six of seven cases with aberrations (86%) showed numerical aberrations of whole chromosomes exclusively. This pattern differed significantly from the findings in melanoma that arose within CMN (n = 6), group VI, or independent of CMN (n = 122) in which only 5% showed numerical changes only. The single case in group V showed aberrations similar to melanoma. The finding of frequent numerical chromosomal aberrations in atypical nodular proliferations arising in CMN identifies these as clonal neoplasms with a genomic instability consistent with a mitotic spindle checkpoint defect. This difference compared to the aberration pattern found in melanoma might explain their more benign clinical behavior and may be of diagnostic value in ambiguous cases.     

ST2作为Th2细胞亚群标志以及其与支气管哮喘的关系的研究

各自主要分泌IFN γ和IL 4的Th1和Th2亚群 ,与临床疾病的关系十分密切。如何从表面标志上加以区分是一项迫切需要解决的问题。ST2是近年来提出的Th2细胞的稳定标志物。本工作在体外成功地诱导人脐带血T细胞向Th1或Th2分化的基础上 ,应用逆转录PCR分析了ST2mRNA的表达特点。证实ST2在人Th2细胞上的选择性表达。为了探索ST2、Th2与支气管哮喘的关系 ,本工作进一步检测了正常人和支气管哮喘患者外周血单个核细胞中 β actin、ST2以及IFN γ和IL 4的mRNA水平。结果显示 :支气管哮喘患者ST2mRNA水平升高 ,IL 4水平也明显升高 ,但IFN γ无变化。这提示ST2作为Th2细胞的标志物 ,有可能成为Th2极化性疾病如哮喘发病机制研究的一个参考性标志 ,至于ST2是否有可能作为治疗的靶分子 ,有待进一步探讨     

Rises in antibody to human herpesvirus 6 detected by enzyme immunoassay in transplant recipients with primary cytomegalovirus infection.

Immunoglobulin G to human herpesvirus 6 (HHV-6) and cytomegalovirus (CMV) in sera from solid organ recipients was measured by an enzyme-linked immunoassay (ELISA) before and after transplant. The HHV-6 ELISA was developed from glycine extracts of HHV-6-infected and uninfected HSB-2 cells. At a serum dilution of 1:500, 80 (91%) of 88 recipients were seropositive for HHV-6 before transplant, while only 14 (16%) were seropositive for CMV. Posttransplant HHV-6 serologic rises were observed in 38 (43%) recipients; rises in 25 of these recipients were associated with primary CMV infection. Titration of sera revealed much higher HHV-6 titer rises among those with primary CMV infection than among those with CMV reactivation or with no CMV infection. Elevated HHV-6 antibody titers persisted for up to 2 years after primary CMV infection. No correlation was noted between CMV and HHV-6 antibody titers in individual serum samples.