A multiplex polymerase chain reaction for the simultaneous detection of the virus and satellite components associated with cotton leaf curl begomovirus disease complex.

Cotton leaf curl disease (CLCuD) is caused by a complex of several whiteflies (Bemisia tabaci Genn.)-transmitted begomovirus species, Cotton leaf curl Multan virus (CLCuMuV), Cotton leaf curl Kokhran virus (CLCuKoV) and Cotton leaf curl Alabad virus (CLCuAlV) by individual of mixed infection, associated with Cotton leaf curl Multan betasatellite (CLCuMB) and several alphasatellites. The disease causes major economic losses in cotton in the Indian subcontinent. For monitoring of epidemiology and development of management strategies of CLCuD, a quick, sensitive and effective method capable of detecting all the begomovirus, betasatellite and alphasatellite components associated with CLCuD is required. With this objective, a multiplex polymerase chain reaction (mPCR) assay was developed for the simultaneous detection of these three viral components associated with CLCuD of cotton. Primers for each component were designed based on the retrieved reference sequences from the GenBank. Each pair of primers, designed for each of the respective component, was evaluated for its sensitivity and specificity in both the component-specific simplex polymerase chain reaction (sPCR) and mPCR assay. This report identified three viral component-specific pairs of primers which, in all combinations, amplified simultaneously the CP gene (780 nts) of the begomovirus, the ßC1gene (375 nts) of the betasatellite and the Rep gene (452 nts) of the alphasatellite associated with CLCuD in the mPCR assays. The amplified products specific to each component produced by these assays were identified based on their amplicon sizes, and the identities of the viral components amplified were confirmed by cloning and sequencing the amplicons obtained in the mPCR. The mPCR assay was validated using naturally CLCuD-affected cotton plants of the fields. This assay will be useful for rapid detection of CLCuD-associated begomovirus, betasatellite and alphasatellite DNA in field samples, extensive resistance screening in resistance breeding programme, and also monitoring epidemiology for detection of virus and its components when symptoms are mild or absent in the plant.

Severe intravascular hemolysis associated with brown recluse spider envenomation. A report of two cases and review of the literature.

Envenomation by the brown recluse spider (loxoscelism) is classically associated with a necrotic ulcer. Systemic manifestations occur in a minority of cases, but are generally mild and self-limited. The hematologic complications of brown recluse spider bite range from mild hemolysis to fulminant intravascular hemolysis with or without evidence of disseminated intravascular coagulation. Intravascular hemolysis is a rare but occasionally lethal complication of brown recluse spider envenomation. This article presents two cases of severe hemolysis associated with loxoscelism occurring in two young women in Memphis, Tennessee. The second documented death in an adult from severe hemolysis due to a brown recluse spider bite is reported. A review of the literature emphasizing the pathogenic mechanisms of spider bite hemolysis is also included.

Association of Chlamydia trachomatis immunoglobulin gamma titers with dystrophic peritoneal calcification, psammoma bodies, adhesions, and hydrosalpinges.

OBJECTIVE: To correlate Chlamydia trachomatis immunoglobulin gamma (IgG) titers with psammoma bodies, dystrophic peritoneal calcification, degree of calcification, adhesions, and hydrosalpinges. DESIGN: This is a prospective single-blinded histologic analysis of tissue and retrospective analysis of historical laboratory and clinical variables. SETTING: Tertiary hospital and private practice patient charts. PATIENTS: Sixty consecutive patients with C. trachomatis IgG titers reported on the coding sheets of a previous study for endometriosis. MAIN OUTCOME MEASURES: The histologic slides were reviewed in a blinded fashion for calcification. Previously used data sheets were reviewed for C. trachomatis IgG titers. Historical data, adhesion scores, hystrosalpingogram findings, and laparoscopic findings were obtained from charts. RESULTS: Dystrophic calcification, psammoma bodies, moderate-to-severe dystrophic calcification and hydrosalpinges were associated with positive C. trachomatis IgG titers. CONCLUSION: This study suggests relationship of C. trachomatis with dystrophic calcification, psammoma bodies, adhesions, and hydrosalpinges. This relationship suggests that C. trachomatis IgG titers can be used as a marker to help determine those infertility patients who might best benefit from hysterosalpingogram or laparoscopy and in clinical studies of endometriosis, infertility, pain, or ovarian cancer. However, there is no current data to suggest a need for therapy on the basis of a positive C. trachomatis IgG titer or of dystrophic peritoneal calcification.

Placental metastasis from maternal carcinoma. A report of three cases.

Metastasis of a maternal neoplasm to the products of conception is extremely rare. Of the 54 reported cases in the world literature, only 14 (25%) showed fetal metastasis. More than 50% of the reported cases were not examined grossly or had no visually apparent tumor deposits. Malignant melanoma is the most common malignant maternal neoplasm to metastasize to the products of conception. We report three unusual maternal malignant neoplasms (one pancreatic and two breast cancer) with evidence of placental metastasis and discuss the risk factors for fetal involvement in these cases.

Anecdotal association of endosalpingiosis with chlamydia trachomatis IgG titers and Fitz-Hugh-Curtis adhesions.

STUDY OBJECTIVE: To correlate Chlamydia trachomatis IgG titers with endosalpingiosis. DESIGN: Prospective, single-blind, histologic analysis of tissue, and retrospective analysis of historical laboratory and clinical variables. SETTING: A tertiary care patient practice. PATIENTS: Sixty consecutive patients with C. trachomatis IgG titers, 28 positive and 32 negative. INTERVENTIONS: The second review of histology slides and charts of these women. MEASUREMENTS AND MAIN RESULTS: One patient was found to have endosalpingiosis. She had Fitz-Hugh-Curtis (FHC) adhesions and a positive C. trachomatis IgG titer, but no endometriosis. CONCLUSIONS: This review confirmed an anecdotal association of endosalpingiosis with positive C. trachomatis IgG titers and FHC adhesions, but failed to demonstrate a statistically significant relationship due to the size of the sample.

Buttonhole ulceration and perforation of the rectum.

Three laparoscopies were performed with treatment of cul-de-sac endometriosis near the rectum Two patients had a subsequent laparotomy for rectal perforation. The bowel damage appears to have been from a combination of failure to appreciate the position of the bowel with anatomy distorted by endometriosis, dissecting into healthy tissue with a carbon dioxide laser, desiccating healthy tissue with bipolar electrosurgery, and thermal spread into healthy tissue from the desiccated endometriotic tissue. Gynecologists must be aware of this potential complication when treating endometriosis near the rectum.

The neuropeptide/mast cell secretagogue substance P is expressed in cutaneous melanocytic lesions.

Substance P (SP) is a neuropeptide found in both the central and peripheral nervous system. In the skin, SP-containing neurons stimulate the release of histamine from connective tissue mast cells (MC). SP also can potentiate neoangiogenesis and induce dermal fibrosis. MC-derived histamine has potent vasoactive effects, is angiogenic, and promotes tissue fibroplasia. In addition to histamine, MC contain many other angiogenic factors and a variety of cytokines, growth factors, and proteolytic enzymes implicated in tissue remodeling, and normal as well as tumor-associated neoangiogenesis. Many MC-derived factors, including histamine, can enhance melanoma cell growth directly. MC often concentrate around cutaneous melanomas which also frequently are associated with angiogenesis and peritumoral fibrosis. The precise mediators of these responses have not been well defined. We evaluated by immunohistochemistry cutaneous lesions representing stages of progression of malignant melanoma and its precursor lesions for the expression of SP. SP was expressed in 17/25 (68%) primary invasive malignant melanomas, 2/5 (40%) metastatic melanomas, 6/10 (60%) melanomas in situ, 7/12 (58%) atypical (dysplastic) nevi, and 4/10 (40%) spindle and epithelioid cell (Spitz) nevi, but was not detected in any (0/11, 0%) acquired benign melanocytic nevi (p<0.05). Invasive melanomas were immunolabeled in both the intraepidermal and the dermal components of the lesions. For those atypical and Spitz nevi which expressed SP, most of the immunoreactive melanocytes were located at the dermal-epidermal junction overlying areas of papillary dermal fibrosis. The results show differential expression of SP among cutaneous melanocytic lesions and suggest that the expression of this neuropeptide together with other factors may contribute to some of the host responses associated with these lesions.

A comparative study of ovarian and pelvic wall-infiltrating endometriosis.

STUDY OBJECTIVE: To compare the histology of ovarian endometriomas (OE) with that of pelvic wall-infiltrating endometriosis (PWIE) in an attempt to elucidate their histopathologic and clinical differences. DESIGN: Retrospective histologic study. SETTING: University-affiliated hospital. SUBJECTS: Randomly selected tissue samples of 10 cases each of OE and PWIE. Interventions. Immunoperoxidase and Masson's trichrome stains were applied to the samples to determine the presence of myofibroblasts and collagen, respectively. MEASUREMENTS AND MAIN RESULTS: All cases of OE showed changes suggestive of type 3 collagen deposition and myofibroblast proliferation. All cases of PWIE showed changes suggestive of type 1 collagen deposition and extensive smooth muscle metaplasia. The endometrial glands and stroma of OE tended to be on the inside lining of the chocolate cyst and did not freely intermix with native ovarian collagen. The lesions of PWIE, however, showed endometrial glands and stroma mixing freely with the native pelvic wall collagen. CONCLUSIONS: These findings suggest two different mechanisms for the pathogenesis of OE and PWIE and offer a rationale for using different surgical approaches to the different sites. Ovarian endometriosis is a lesion within the ovary with a response compatible with recent metastasis, whereas PWIE appears to be a metaplastic reaction.

Use of algorithmic pathways to develop quality, cost-effective clinical care.

STUDY OBJECTIVES: To analyze the cost of diagnosis and treatment associated with hydrosalpinges and pelvic adhesions using algorithmic pathways. DESIGN: Analysis of six pathways: (1) Chlamydia IgG antibody titer -> hysterosalpingogram -> diagnostic laparoscopy -> in vitro fertilization (IVF); (2) Chlamydia IgG antibody titer -> diagnostic laparoscopy -> therapeutic laparoscopy -> IVF; (3) hysterosalpingogram -> diagnostic laparoscopy -> therapeutic laparoscopy -> IVF; (4) diagnostic laparoscopy -> therapeutic laparoscopy -> IVF; (5) no treatment; and (6) IVF only. SETTING: A tertiary care practice at a university-associated hospital. PATIENTS: Patients were retrospectively chosen on the basis of availability of results of Chlamydia IgG titers, operative notes from laparoscopy, and hysterosalpingograms (HSGs). All patients identified with these three results available were included. INTERVENTIONS: Determination of results of laparoscopic treatment, IVF, and no therapy (observation only), based on the literature. MEASUREMENTS AND MAIN RESULTS: The theoretical cost to achieve pregnancy for each pathway was calculated by mathematical modeling. The greatest number of pregnancies for the least cost ($18, 883/pregnancy) was from the diagnosis and treatment of adhesions at laparoscopy with no previous screening. The least costly approach to pregnancy for blocked tubes ($26,614/pregnancy) was to start with an HSG. All pathways for adhesions and any screening pathway using HSG for hydrosalpinges were more cost effective than IVF. CONCLUSIONS: These pathways are limited, as they require several assumptions and are based on a tertiary care population. Although HSG and laparoscopy appeared to be the most cost-effective approaches to pregnancy, a primary care group may have a more cost-effective response to the use of Chlamydia trachomatis IgG antibody screening.

Association of positive Chlamydia trachomatis and Chlamydia pneumoniae immunoglobulin-gamma titers with increasing age.

STUDY OBJECTIVES: To use Chlamydia trachomatis immunoglobulin-gamma (IgG) titers to investigate the possibility of their association with ovarian cancer, and to evaluate the effectiveness of this titer in algorithmic protocols in infertility. DESIGN: Prospective, age-matched, pilot study (Canadian Task Force classification II-2). SETTING: University and university-affiliated office practice. PATIENTS: The original 30 patients were seen for follow-up of ovarian cancer (19) and at yearly examination for nonmalignant disease (10). An additional group of 21 women seen for pelvic pain and infertility was added to clarify questions that arose during the study. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Chlamydia trachomatis IgG titers were positive in 15 (79%) of 19 of women with ovarian cancer, 9 (90%) of 10 age-matched controls, and 14 (67%) of 21 patients with infertility and pain. When analyzed by age, 4 (40%) of 10 patients under 30 years and 34 (85%) of 40 patients 30 years of age or older had positive titers (p = 0.007). Of 21 women with positive Chlamydia pneumoniae titers, 17 (81 %) had positive C. trachomatis titers, and 17 (85%) of 20 with positive C. trachomatis titers had positive C. pneumoniae titers. CONCLUSION: The test kit used in this study may not be adequate in older patients due to cross-reaction with C. pneumoniae titers. Further evaluation of C. trachomatis IgG titers as a marker in the study of ovarian cancer will require titers that are more specific than those we used. Although these titers may be useful as an immunologic screening marker in infertile patients, results should be interpreted with caution. A positive test may not be evidence of C. trachomatis infection and is not an indication for specific therapy. Successful use of some currently available C. trachomatis IgG titers in algorithms for infertility may be related to a patient's age.

Non-hematopoietic cutaneous metastases in children and adolescents: thirty years experience at St. Jude Children's Research Hospital.

BACKGROUND: The spectrum of cutaneous metastasis of non-hematopoietic neoplasms in the pediatric population is not well documented. We report the histologic diversity of this unusual process over a 30-year period at a tertiary care center for pediatric malignancy (St. Jude Children's Research Hospital, Memphis, TN, USA). METHODS: Of 1,971 pathology accessions which included histologic material on skin (1,604 surgical cases and 367 autopsy cases) we found 40 cases (2% of total skin accessions) coded for metastatic non-hematopoietic malignancy. RESULTS: The patients (n=34) ranged in age from 1 month to 20 years (mean=9.8 years) and had a male:female ratio of 1:1. The histologic diagnoses were as follows: rhabdomyosarcoma NOS (6 cases), embryonal rhabdomyosarcoma (4 cases), alveolar rhabdomyosarcoma (4 cases), neuroblastoma (8 cases), osteosarcoma (2 cases), choriocarcinoma (2 cases), peripheral neuroepithelioma or Ewing's sarcoma (2 cases), malignant rhabdoid tumor (1 case), paraganglioma (1 case), nasopharyngeal carcinoma (1 case), sarcoma NOS (1 case), colon adenocarcinoma (1 case), and malignant melanoma (1 case). CONCLUSIONS: Cutaneous or subcutaneous metastasis of non-hematopoietic malignancies in children and adolescents is a rare occurrence but in a high percentage of cases may be the first manifestation of disease. The tumors most likely to metastasize to the skin in children are rhabdomyosarcoma and neuroblastoma and they are more likely than adult malignancies to disseminate to multiple distant sites.