Limited genetic variability of Cytauxzoon felis apical membrane antigen-1 (ama1) from domestic cats and bobcats.

BACKGROUND: Cytauxzoon felis is a tick-transmitted apicomplexan that causes cytauxzoonosis in domestic cats (Felis catus). Even with intensive care, the mortality rate of acute cytauxzoonosis approaches 40% in domestic cats, while bobcats (Lynx rufus), the natural intermediate host of C. felis, remain clinically asymptomatic. However, multiple reports of domestic cats surviving acute disease without any treatment exist. One hypothesis for survival of these cats is infection with unique C. felis genotypes of lower pathogenicity. Prior studies have identified genetically distinct C. felis isolates containing polymorphisms within internal transcribed spacer regions (ITS) of the rRNA operon. However, these polymorphisms do not correlate with the clinical outcome of cytauxzoonosis, and so additional genetic markers are needed to test this hypothesis. We selected C. felis apical membrane antigen-1 (ama1) as a potential genetic marker of differential pathogenicity. AMA1 is a vaccine candidate for relatives of C. felis within Plasmodium spp.; however its historically high level of genetic polymorphism has resulted in escape from vaccine-induced immunity. While such diversity has hindered vaccine development, the expected polymorphism within the ama1 gene may be useful to evaluate population genetics. RESULTS: A 677 bp sequence of the C. felis ama1 gene was PCR-amplified from 84 domestic cats and 9 bobcats and demonstrated 99.9% sequence identity across all samples. A single nucleotide polymorphism (SNP) was identified in domestic cats and bobcats with evidence for co-infection with both genotypes identified in two domestic cats. The prevalence of the two genotypes varied with geographical distribution in domestic cats. Nucleotide diversity (π) and haplotype diversity (H) were calculated for C. felis ama1 and ama1 of related apicomplexans to assess genetic diversity. Based on these values (π = 0.00067 and H = 0.457), the diversity of the C. felis ama1 gene region analyzed is considerably lower than what is documented in related apicomplexans. CONCLUSIONS: In surprising contrast to related apicomplexans, our results support that the sequence of the C. felis ama1 gene is highly conserved. While lack of genetic diversity limits utility of C. felis AMA1 as a genetic marker for clinical outcome, it supports further investigation as a vaccine candidate for cytauxzoonosis.

Atypical cytologic presentation of a histiocytic sarcoma in a Cavalier King Charles Spaniel dog.

A Cavalier King Charles Spaniel dog was presented because of a 10-d history of progressive vomiting, inappetence, and lethargy, with mild neurologic signs. Fine-needle aspirates of splenic nodules seen on ultrasound were suggestive of a carcinoma. On autopsy, a disseminated neoplasm was present in the lung, spleen, and adrenal glands. Additionally, there was a Chiari-like malformation of the skull with corresponding syringomyelia of the cranial spinal cord. Histologically, the neoplasm was comprised of a pleomorphic population of round cells with a high mitotic rate. Cells stained positive for vimentin and CD18 and negative for cytokeratin, chromogranin A, CD3, CD20, MHC II, and CD11d, and hence the tumor was diagnosed as a histiocytic sarcoma. The cytologic presentation in this case is uncommon for this type of tumor, as was the clinical presentation.

MANDIBULAR SQUAMOUS CELL CARCINOMA IN A BOBCAT (LYNX RUFUS).

A 23-yr-old female spayed bobcat (Lynx rufus) presented with a 1-wk history of hypersalivation. On examination, the right mandible was markedly thickened, the right mandibular dental arcade was missing, and the oral mucosa over the right mandible was ulcerated and thickened. Skull radiographs and fine needle aspirate cytology were supportive of squamous cell carcinoma. The bobcat was euthanized as a result of its poor prognosis. Necropsy confirmed a diagnosis of oral squamous cell carcinoma of the mandible. To the authors' knowledge, this is the first report of oral squamous cell carcinoma in a bobcat.

Identifying postpartum depression: are 3 questions as good as 10?

BACKGROUND: Postpartum depression is the most common medical problem that new mothers face. Anxiety is a more prominent feature of postpartum depression than of depression that occurs at other times in life. Routine, universal screening significantly improves detection in primary health care settings. Thus, an ultrabrief scale that could be incorporated into a general health survey or interview would be useful. OBJECTIVE: We tested the hypothesis that, during the first 6 postpartum months, the 3-item anxiety subscale of the Edinburgh Postpartum Depression Scale is a better ultrabrief depression screener than 2 Edinburgh Postpartum Depression Scale questions that are almost identical to the widely used Patient Health Questionnaire. METHODS: A cohort of 199 14- to 26-year-old participants in an adolescent-oriented maternity program completed the Edinburgh Postpartum Depression Scale at well-child visits during the first 6 postpartum months. Three subscales of the Edinburgh Postpartum Depression Scale were examined as ultrabrief alternatives: the anxiety subscale (3 items; Edinburgh Postpartum Depression Scale-3), the depressive symptoms subscale (7 items; Edinburgh Postpartum Depression Scale-7), and 2 questions that resemble the Patient Health Questionnaire (Edinburgh Postpartum Depression Scale-2). The reliability, stability, and construct validity of the Edinburgh Postpartum Depression Scale and 3 subscales were compared. Criterion validity was assessed by comparison with a score of >/=10 on the full, 10-item Edinburgh Postpartum Depression Scale. RESULTS: A total of 41 mothers (20.6%) met study criteria for referral for evaluation of depression (Edinburgh Postpartum Depression Scale-10 score >/= 10). The Edinburgh Postpartum Depression Scale-3 exhibited the best screening performance characteristics, with sensitivity at 95% and negative predictive value at 98%. It identified 16% more mothers as depressed than the Edinburgh Postpartum Depression Scale did. The performance of the Edinburgh Postpartum Depression Scale-2 was markedly inferior, with sensitivity at 48% to 80%. Moreover, the Edinburgh Postpartum Depression Scale-2 was unreliable for mothers who had not been depressed in the past. CONCLUSION: The brevity, reliability, and operating characteristics of the Edinburgh Postpartum Depression Scale-3 make it an attractive postpartum depression screening tool for primary health care settings in which the goal is to detect depression, not to assess its severity. Validation by diagnostic psychiatric interview is needed.

Association of conventional goals and perceptions of pregnancy with female teenagers' pregnancy avoidance behavior and attitudes.

CONTEXT: Fostering conventional goals is a key component of pregnancy prevention interventions for teenagers. However, research has not shown whether having goals independently influences sexual behavior, or whether the perception that pregnancy represents an impediment to achieving goals mediates any association. METHODS: In 1999-2001, a racially mixed group of 351 sexually experienced female teenagers who were inadequate contraceptive users completed surveys about goals, the anticipated impact of childbearing on these goals, and protective behaviors and attitudes. Chi-square, regression and two-by-two table analyses assessed associations between goals and perceptions of early childbearing and pregnancy avoidance measures. RESULTS: Three-fourths of respondents had educational or vocational goals. Eight in 10 of these teenagers perceived their goals to be achievable, but fewer than half thought pregnancy would be an impediment to achieving these goals. Teenagers who had goals were more likely than others to have used a contraceptive at last intercourse (odds ratio, 1.9), but controlling for the perception of pregnancy as an impediment eliminated this association. In contrast, considering pregnancy an impediment was associated with an increased likelihood of supporting each pregnancy avoidance measure (2.1-9.6), and of intending to avoid pregnancy and to have an abortion if pregnant, regardless of whether teenagers had goals (8.3-13.8). CONCLUSIONS: Conventional goals appear to motivate teenagers to avoid getting pregnant only if they believe pregnancy will be an impediment. Thus, it may be less important to encourage young women to formulate goals than to ensure that they consider adolescent childbearing a threat to their plans.

Feasibility and utility of screening adolescent mothers for Chlamydia at their children's health care visits.

PURPOSE: Assess the feasibility of offering and barriers to accepting urine-based screening for Chlamydia trachomatis (CT) among asymptomatic adolescent mothers during their children's health care visits. METHOD: Providers were automatically cued to offer CT-screening to 13 through 21 year old mothers when they opened the child's medical record. Recording the mothers' screening decisions removed the flag for 6 months. Providers were also prompted to assess the perceived importance of CT prevention, likelihood of having CT, and the worst aspect of having CT. RESULTS: Mothers usually brought their children to the clinic. Hence, providers could act on 97% of the 318 flags they saw. They responded appropriately 75% of the time. Only 96 (42%) of the 229 mothers who were asked agreed to screening. The primary reasons for declining were "monogamous" and "tested within 6 months". Almost everyone said CT was a top preventive health priority, but that they would be surprised if they were infected. Mothers who agreed to screening were less likely to have a child less than 6 months of age (26.0% vs. 57.1%; P < 0.0001). They were also more likely to rate knowledge of partner infidelity (39.4% vs. 13.6%; P = 0.03) and less likely to rate medical problems (15.2% vs. 40.9%; P = 0.01) the worst aspect of having CT. None of the 21 urine samples obtained within 6 months of delivery were positive for CT. Thereafter, 5 (9.1%) of the 55 urines were positive. CONCLUSIONS: It is feasible and useful to screen adolescent mothers for CT at their children's health care visits. The best way to increase the efficacy and strengthen the impact of urine-based CT-screening in this setting is to initiate testing after the 6th postpartum month and heighten awareness of STD-risk among adolescent mothers.

Determination of hyperglycosylated human chorionic gonadotropin produced by malignant gestational trophoblastic neoplasias and male germ cell tumors using a lectin-based immunoassay and surface plasmon resonance.

The ability to reliably detect aberrant glycosylation of human chorionic gonadotropin (hCG) may have profound implications for the diagnosis and monitoring of malignant gestational trophoblastic neoplasia, germ cell tumors, other malignancies, and pregnancy complications. To become a clinically useful assay, however, this discrimination of glycoforms should be possible on minimally treated biological specimens. Towards this end, we have developed a lectin-based sandwich-type immunoassay to compare the glycosylation patterns of hCG among urine specimens from patients presenting with a normal pregnancy, invasive mole, choriocarcinoma, and male germ cell tumors using carbohydrate-free antibody fragments as capture reagents and a panel of eight lectins, five recognizing neutral sugars and three recognizing sialic acid. There was no significant difference in the binding of any of the lectins to hCG in the urine of women over the gestational range of 6-38 weeks. Three lectins, however, exhibited differential binding to urinary hCG derived from these normal pregnant controls and that from patients with malignant forms of gestational trophoblastic disease and male germ cell tumors. Galanthus nivalis agglutinin and Maackia amurensis lectin, which bind terminal mannose and alpha(2-3)sialic acid, respectively, preferentially bound pregnancy-derived hCG, whereas the lectin, wheat germ agglutinin, which binds sialic acid and beta(1-4)N-acetylglucosamine, exhibited decreased binding to pregnancy-derived hCG compared to that from patients with male germ cell tumors and malignant gestational trophoblastic neoplasia. The differential binding observed with these three promising lectins is most encouraging and warrants further examination. The experimental paradigm also holds promise for the development of comparable assays for other glycosylated tumor markers.

Why lightning strikes twice: postpartum resumption of sexual activity during adolescence.

PURPOSE: To describe the circumstances surrounding the postpartum resumption of sexual activity in a cohort of teenage mothers. The goal was to shed new light on the reasons why teenagers who have just given birth put themselves at risk for conception. METHODS: A racially and ethnically diverse group of 267 poor, predominantly unmarried, primiparous, 13-21 year olds was enrolled consecutively at delivery and followed through the third postpartum month. The primary outcome measure was weeks postpartum at resumption of sexual activity, further categorized as protected or unprotected. The analysis controlled for factors thought to influence the postpartum resumption of sexual activity among adult couples and the use of contraception during adolescence. The Kaplan-Meier method was used to estimate the time to end point and Cox proportional-hazards regression analysis to compute relevant relative risks (RR). RESULTS: By the end of the third postpartum month, 58% of the teenagers had had sexual intercourse and the majority (80%) used contraception when they did so. The median time to first coitus, 10.7 weeks, was unrelated to contraceptive use but was significantly shorter among teenagers who lived with their boyfriends (RR: 2.4; 95%CI: 1.7-3.4) and those who delivered prior to term (RR: 2.1; 95%CI: 1.3-3.6). The analysis revealed that the teenagers who did not use contraception at first postpartum sexual intercourse exhibited more theorized risk factors for conception than those who did and those who remained sexually abstinent. CONCLUSIONS: This new empirical evidence that coital activity resumes soon after delivery should dispel the normative belief that contraception is unnecessary during the puerperium. Early contraceptive vigilence may also decrease the frequency and rapidity with which teen mothers conceive, as the differences in the prevalence of teen pregnancy risk factors we uncovered suggest that decisions about using contraception (not sexual intercourse) determine the risk of rapid repeat conception. Teens with live-in boyfriends and premature babies are especially apt to benefit from the new information in this report as they resume sexual activity sooner than their peers and are less apt to use contraception when they do so.

Lectin immunoassays using antibody fragments to detect glycoforms of human chorionic gonadotropin secreted by choriocarcinoma cells.

Immobilized antibodies are commonly used to recognize and bind proteins of interest from heterogeneous samples; however, subsequent probing of the glycan(s) of captured glycoproteins with lectins is limited by interference due to the competing oligosaccharides inherently present on antibodies. To prepare capture antibodies with significantly reduced binding of any lectin, the glycosylated protein domains (F(c)) of two anti-human chorionic gonadotropin antibodies were proteolytically removed. Depending on the individual antibody, usable fragments were generated either directly or effectively separated after cleavage through partial reduction and thiol coupling to an appropriate matrix. Importantly, neither method required additional purification of the antibody fragments before immobilization. Binding of a variety of lectins to the functional fragments was reduced by approximately 90% compared with intact immunoglobulin G in both an enzyme-linked immunosorbent assay and a biosensor format. These carbohydrate-free antibody fragments were used to bind the glycoprotein hormone, human chorionic gonadotropin, produced during normal pregnancy and that secreted by three human choriocarcinoma cell lines. Lectins bound to the unpurified gonadotropin glycoforms in distinct patterns consistent with glycan structures previously elucidated by others on hormone samples purified from the urine of pregnant women and of patients with choriocarcinoma. The methods described in this article are applicable for generating capture reagents universally suitable for lectin immunoassays of glycoproteins.