ARX polyalanine expansion mutations lead to migration impediment in the rostral cortex coupled with a developmental deficit of calbindin-positive cortical GABAergic interneurons.

The Aristaless-related homeobox gene (ARX) is indispensable for interneuron development. Patients with ARX polyalanine expansion mutations of the first two tracts (namely PA1 and PA2) suffer from intellectual disability of varying severity, with seizures a frequent comorbidity. The impact of PA1 and PA2 mutations on the brain development is unknown, hindering the search for therapeutic interventions. Here, we characterized the disturbances to cortical interneuron development in mice modeling the two most common ARX polyalanine expansion mutations in human. We found a consistent ∼40-50% reduction of calbindin-positive interneurons, but not Stt+ or Cr+ interneurons, within the cortex of newborn hemizygous mice (p=0.024) for both mutant strains compared to wildtype (p=0.011). We demonstrate that this was a consequence of calbindin precursor cells being arrested or delayed at the ventral subpallium en route of tangential migration. Ex-vivo assay validated this migration deficit in PA1 cells (p=0.0002) suggesting that the defect is contributed by intrinsic loss of Arx function within migrating cells. Both humans and mice with PA1 mutations present with severe clinical features, including intellectual disability and infantile spasms. Our data further demonstrated the pathogenic mechanism was robustly shared between PA1 and PA2 mutations, as previously reported including Arx protein reduction and overlapping transcriptome profiles within the developing mouse brains. Data from our study demonstrated that cortical calbindin interneuron development and migration is negatively affected by ARX polyalanine expansion mutations. Understanding the cellular pathogenesis contributing to disease manifestation is necessary to screen efficacy of potential therapeutic interventions.

Matrix-assisted laser desorption/ionisation-time of flight mass spectrometry: rapid identification of bacteria isolated from patients with cystic fibrosis.

Despite extensive research into the diagnosis and management of cystic fibrosis (CF) over the past decades, sufferers still have a median life expectancy of less than 37 years. Respiratory tract infections have a significant role in increasing the morbidity and mortality of patients with CF via a progressive decline in lung function. Rapid identification of organisms recovered from CF sputum is necessary for effective management of respiratory tract infections; however, standard techniques of identification are slow, technically demanding and expensive. The aim of this study is to asses the suitability of matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI-TOF MS) in identifying bacteria isolated from the respiratory tract of patients with CF, and is assessed by testing the accuracy of MALDI-TOF MS in identifying samples from a reference collection of rare CF strains in conjunction with comparing MALDI-TOF MS and standard techniques in identifying clinical isolates from sputum samples of CF patients. MALDI-TOF MS accurately identified 100% of isolates from the reference collection of rare CF pathogens (EuroCare CF collection). The isolate identification given by MALDI-TOF MS agreed with that given by standard techniques for 479/481 (99.6%) clinical isolates obtained from respiratory samples provided by patients with CE In two (0.4%) of 481 samples there was a discrepancy in identification between MALDI-TOF MS and standard techniques. One organism was identified as Pseudomonas aeruginosa by MALDI-TOF but could only be identified by the laboratory's standard methods as of the Pseudomonas genus. The second organism was identified as P. beteli by MALDI-TOF MS and Stenotrophomonas maltophilia by standard methods. This study shows that MALDI-TOF MS is superior to standard techniques in providing cheap, rapid and accurate identification of CF sputum isolates.

First Report of Glomerella cingulata (Colletotrichum gloeosporioides) Causing Anthracnose and Tip Dieback of Lygodium microphyllum and L. japonicum in Australia.

The Old World climbing fern, Lygodium microphyllum (Cav.) R. Br., and Japanese climbing fern, L. japonicum (Thunb.) Sw., are invasive noxious weeds in Florida (1). Exploratory surveys for classical biological control agents of L. microphyllum in the fern's native range of Australia and Asia have focused on aboveground herbivores (1). From February to August 2006, fungi were isolated from symptomatic foliage, including lesions associated with leaf curls caused by the mite Flocarus perrepae Knihinicki & Boczek., obtained from L. microphyllum at sites across southeast Queensland, Australia and from both fern species grown at the CSIRO Long Pocket Laboratories in Brisbane, Australia. Anthracnose symptoms with chlorotic margins, initiating at the tip or base of the individual pinnules, were observed on fronds. Dieback symptoms affected growing tips, with sunken lesions and a gradual necrotic wilt as far as the next growth junction of pinnae. Sections from diseased margins were surface sterilized, placed onto water agar, and incubated at 23°C with a 16-h photoperiod. Variable colonies of white-to-gray mycelia, felted or tufted with complete margins, grew well on oatmeal agar and potato dextrose agar. Conidia were hyaline to light salmon, aseptate, straight, and cylindrical (10.4 to 18.2 × 2.6 to 5.2 µm), borne in salmon-to-bright orange masses at 25°C, and consistent with previous descriptions of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. (3), anamorph of Glomerella cingulata (2). Asci that formed after 3 to 4 weeks in culture were eight-spored, clavate to cylindrical (46.8 to 62.4 × 9.1 to 11.7 µm), and thickened at the apex, and ascospores were cylindrical (11.7 to 18.2 × 3.9 to 5.2 µm), slightly curved, unicellular and hyaline, which is consistent with descriptions of G. cingulata (2). No fruiting bodies were observed in planta; acervuli, setae, and perethecia were not observed. Identification was further confirmed by molecular analysis using the primer pair ITS1/ITS4 (4) (GenBank Accession No. EU697014), indicating 100% similarity to isolates of G. cingulata. To confirm pathogenicity, Koch's postulates were performed on three plants of L. japonicum and 12 plants of L. microphyllum, with an equal number of controls. Conidial suspensions were made to 1.7 × 106 conidia ml-1. During the experiments in the glasshouse, temperatures ranged from 12.6 to 40°C and relative humidity from 39 to 85%. Tips and fronds were collected after 2 to 8 weeks and isolation and identification performed. G. cingulata was consistently reisolated from diseased tissue. No symptoms appeared on controls and isolations did not yield the pathogen. To our knowledge, this is the first report of G. cingulata infecting L. microphyllum and L. japonicum in Australia. Its potential as a biological control agent in the ferns' introduced range remains to be tested. References: (1) J. A. Goolsby et al. Biol. Control. 28:33, 2003. (2) J. E. M. Mordue. Glomerella cingulata. No. 315 in: CMI Descriptions of Pathogenic Fungi and Bacteria. CAB, Kew, UK, 1971. (3) B. C. Sutton. The Genus Glomerella and its Anamorph Colletotrichum. In: Colletotrichum: Biology, Pathology and Control. J. A. Bailey and M. J. Jeger, eds. CAB International, Wallingford, UK, 1992. (4) T. M. White et al. Amplification and Direct Sequencing of Fungal Ribosomal RNA for Phylogenetics. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

Breast-feeding and serum p,p'DDT levels among Mexican women of childbearing age: a pilot study.

For almost 50 years, millions of Mexicans have been directly and/or indirectly exposed to (1,1,1-trichloro-2,2-bis[p-chlorophenyl]ethane) (p,p'DDT). The potential related health outcomes of this exposure are of international concern. The objective of the study was to determine the effect of breast-feeding on serum levels of 1,1-dichloro-2; 2'-bis(p-chlorophenyl)ethylene (p,p'DDE) and p,p'DDT in women of childbearing age who were residents of the state of Morelos, Mexico. In March 1999, we examined a sample of 24 women, ages 21 to 36 years, who came with their children to the outpatient facility of a county Health Center in the state of Morelos. The geometric mean for p,p'DDE was 21.8 ng/ml+/-2.58 (GSD) and 2.9 ng/ml+/-2.84 for p,p'DDT. For each month of breast-feeding we observed similar significant decreases for both p,p'DDE (beta=-0.0403 per log concentration unit, P=0.001) and p,p'DDT (beta=-0.0309, P=0.03) serum levels, adjusted by mothers' age and number of children. The adjusted half-life estimate for p,p'DDE serum levels was 17 months. It was concluded that breast-feeding leads to rapid removal of p,p'DDT from the body (<2 year half-life) compared to nonlactational elimination rates (<5 year half-life). In this population, serum p,p'DDE levels were similar to those found 20 years ago in the United States.

Anticoagulation of children undergoing cardiopulmonary bypass is overestimated by current monitoring techniques.

HYPOTHESIS: Children who undergo cardiopulmonary bypass (CPB) are proportionally more hemodiluted than adults who undergo CPB. Current methods of monitoring high-dose heparin sulfate anticoagulation are dependent on fibrinogen level. Because of the decreased fibrinogen levels in children, current methods of monitoring heparin anticoagulation overestimate their level of anticoagulation. DESIGN: Prospective controlled trial. MAIN OUTCOME MEASURE: Production of thrombin (adequacy of anticoagulation). METHODS: Children and adults undergoing cardiac surgery who received CPB were anticoagulated in the standard fashion as directed by activated clotting time (ACT) results. Each subject had blood sampled at baseline; heparinization; start of the CPB; CPB at 30, 60, and 90 minutes; and at termination of CPB. Samples were used to assess anticoagulation with the Heparin Management Test (less dependent on fibrinogen level than ACT). We also assessed 2 subclinical markers of thrombosis, thrombin-antithrombin complexes and prothrombin fragment F1.2; a marker of procoagulant reserve, fibrinogen; the natural antithrombotic, antithrombin; and heparin concentration. RESULTS: Ten children and 10 adults completed the study. Children had lower fibrinogen levels than adults throughout CPB (P<.05). All adults had both therapeutic ACT and Heparin Management Test levels measured throughout CPB. Although children had therapeutic ACT levels, their Heparin Management Test levels were subtherapeutic while undergoing CPB. The children had significantly higher thrombin-antithrombin complexes and prothrombin fragment F1.2 than adults, indicating ongoing thrombin production (P<.01). The increases in thrombin-antithrombin complexes and prothrombin fragment F1.2 in children were inversely proportional to their weight. CONCLUSIONS: Children undergoing CPB with heparin dosing adjusted to optimize the ACT manifest inadequate anticoagulation (ongoing thrombin formation). High-dose heparin anticoagulation therapy in children undergoing CPB should be directed by tests (like the Heparin Management Test) that are less dependent on fibrinogen level than ACT.

Sinus histiocytosis presenting as bilateral epibulbar masses.

PURPOSE: To report a case of sinus histiocytosis presenting as multiple, bilateral epibulbar masses. METHODS: We examined a 4-year-old child with a 2-month history of gradually enlarging, salmon-colored epibulbar masses. Excisional biopsies and a systemic evaluation were performed. RESULTS: Systemic evaluation was normal; no lymphadenopathy was documented. Histopathologic evaluation disclosed features consistent with sinus histiocytosis. Flow cytometry and gene rearrangement studies confirmed the benign nature of the disease. No tumor recurrence was noted. CONCLUSION: Sinus histiocytosis may present as isolated epibulbar masses that may be treated with surgical excision.

Predictors of plasma concentrations of DDE and PCBs in a group of U.S. women.

We evaluated predictors of plasma concentrations of dichlorodiphenyldichloroethylene (DDE), a metabolite of dichlorodiphenyltrichloroethane (DDT), and polychlorinated biphenyls (PCBs) in a group of 240 women, controls from a breast cancer case-control study nested in the Nurses' Health Study. We considered personal attributes such as age, serum cholesterol, region of residence, adiposity, lactation, and dietary intake. DDE levels increased 0.17 ppb/year of age (p = 0.0003), and PCBs increased 0.08 ppb (p = 0.0001). DDE and PCBs increased 0.20 (p = 0.02) and 0.13 ppb (p = 0.001), respectively, per 10 mg/dl serum cholesterol. Women living in the western United States had higher levels of DDE (mean = 11.0 ppb; p = 0.003), and women in the Northeast and Midwest had higher levels of PCBs (mean = 5.6 ppb; p = 0.0002) as compared to women from other parts of the country (mean DDE = 6.3; mean PCBs = 4. 5 ppb). Levels of DDE could not be predicted from consumption of meat, fish, poultry, dairy products, vegetables, fruits, and grains. There was a positive association between fish consumption and PCB concentrations among women in the Northeast and Midwest. Using data from the cases in the nested case-control study to assess the predictive ability of the models, we confirmed that the most reliable predictors of DDE were age and serum cholesterol, and the most important predictors of PCBs were age, serum cholesterol, and residence in the Midwest or Northeast. The null results for the majority of the food variables suggest that specific dietary factors, other than fish, are not currently a substantial contributor to human exposure to DDE and PCBs.

Surgical management of persistent müllerian duct syndrome.

OBJECTIVES: To describe the optimal surgical management of the testes and müllerian duct structures in patients with persistent müllerian duct syndrome. METHODS: We performed a comprehensive Medline literature search regarding the surgical management of persistent müllerian duct syndrome and extracted information regarding the etiology, pathogenesis, and treatment of this disorder. We specifically assessed the risks of retained müllerian structures versus surgical excision of the infantile uterus and fallopian tubes. Using this information, we formulated a comprehensive strategy for the management of patients with persistent müllerian duct syndrome. An illustrative case is described. RESULTS: No malignant degeneration of persistent müllerian structures has been reported. The risk of testicular neoplasia in persistent müllerian duct syndrome approximates the risk of neoplasia in other intra-abdominal gonads. Fertility has rarely been reported although virilization is unaffected. Surgical excision of the infantile uterus and fallopian tubes risks damage to vasa deferentia and the deferential blood supply to the testis. CONCLUSIONS: Surgical excision of persistent müllerian duct structure may result in ischemic and/or traumatic damage to the vasa deferentia and testes. Optimal surgical management is orchiopexy leaving the uterus and fallopian tubes in situ. Meticulous proximal salpingectomy and hysterectomy is indicated only in patients whose müllerian structures limit intrascrotal placement of the tests. Orchiectomy is indicated for testes that cannot be mobilized to a palpable location.

In vitro and in vivo developmental capacity of oocytes from prepubertal and adult sheep.

Development to the blastocyst stage and survival following embryo transfer were assessed for oocytes obtained from prepubertal and adult sheep matured and fertilized in vitro. The rates of maturation, fertilization and cleavage in vitro did not differ significantly between oocytes from prepubertal and adult sheep. The proportion of cleaved zygotes reaching the blastocyst stage was significantly lower for oocytes derived from prepubertal than for those from adult sheep (15.4% and 34.1% respectively). There were no differences in the pregnancy rate and number of lambs born following transfer of blastocyst stage embryos derived from prepubertal and adult sheep to adult recipients. These data show that embryos derived from prepubertal lamb oocytes have reduced developmental potential in vitro but, of those which do reach the blastocyst stage, they have equal capacity to develop to term as embryos derived from adult sheep.

Plexiform neurofibromatosis involving the uterine cervix, endometrium, myometrium, and ovary.

BACKGROUND: Neurofibromatosis involving the female genital tract is unusual. The vulva appears to be most frequently affected, with only rare reports of vaginal, cervical, and ovarian involvement in patients with known von Recklinghausen's disease. CASE: We present a case of diffuse neurofibromatosis involving the cervix, endometrium, myometrium, ovarian cortex, and serosa in a patient with uncontrollable pelvic pain, diagnosed initially as endometriosis. No gross pathology was appreciated, and the histologic features of the small lesions were consistent with plexiform neurofibromas. CONCLUSION: The case presented is an example of diffuse involvement of the female genital tract by neurofibromatosis in a patient without a family history of von Recklinghausen's disease and without other clinical manifestations of the disease.

Epithelial neoplasms of the ovary. An update on current concepts.

New concepts concerning the histologic features of peritoneal implants in serous tumors of low malignant potential and their proposed impact on prognosis are discussed. Endometrioid tumors of low malignant potential, transitional cell carcinoma, and psammocarcinoma are also reviewed.

New developments in sex cord-stromal and germ cell tumors of the ovary.

This article examines unusual and recently described neoplasms such as variants of Sertoli-Leydig cell tumors, monodermal teratomas, and small cell carcinoma, both the hypercalcemic and pulmonic types. Additionally, clues to assist in differentiating metastatic from primary tumors of the ovary are discussed.

Resection of primary hepatic malignant fibrous histiocytoma, fibrosarcoma, and leiomyosarcoma.

Four patients had resection for primary hepatic sarcoma: one with malignant fibrous histiocytoma (MFH), two with poorly differentiated fibrosarcoma, and one with leiomyosarcoma. Age ranged from 40 to 69 years. One patient had a cousin and a grandmother who had died of hepatic tumors. At presentation, all patients had pain; one had tumor rupture, and one had mental changes and hypoglycemia. None had hepatitis or cirrhosis. Results of laboratory evaluation were nonspecific, including normal carcinoembryonic antigen and alpha-fetoprotein levels. Computed tomography showed hypodense masses with enhancement. Angiography showed a hypervascular mass in three patients and an avascular mass in the patient with MFH. Despite large tumors (8 to 32 cm), portal and hepatic veins were not invaded. The pattern of vascularization and lack of venous invasion helps differentiate primary hepatic sarcomas from hepatocellular carcinoma, especially in noncirrhotic patients. All patients had extensive hepatic resections, with one operative death. Immunohistochemical stains of the tumors were positive for vimentin but negative for epithelial markers, differentiating these lesions from other hepatic tumors. The patient with MFH died with recurrence at 10 1/2 months. The patient with the ruptured fibrosarcoma had a second resection and chemotherapy, but died with recurrence at 3 years. The patient with the leiomyosarcoma had a second resection and was disease free at 4 years. Resection of primary hepatic sarcoma is warranted, with potential survival measured in years.

An issue of sensitivity. Use of analgesic gel in catheterizing women.

Catheterisation is an invasive, and often painful procedure for women, yet the use of analgesic gel does not appear to be the norm. It does, however, have a number of benefits for patient and nurse.

Comparative cytologic yield and quality of three Pap smear instruments.

BACKGROUND: Various sampling techniques of the cervix have established false negative rates ranging from 18% to 45%. A number of studies suggest that this false negative rate can be reduced by sampling techniques that are associated with higher yields of endocervical cells. METHOD: This study enrolled 301 women, each of whom had a Pap smear obtained using three different sampling instruments (a cervical brush, a plastic spatula, and a cotton swab) in random order. RESULTS: The cervical brush yielded a better endocervical sample than either of the other instruments (P < .001), and there was no difference between the swab and spatula. In addition, the recovery of endocervical cells increased with each additional sample taken, regardless of instrument used (P < .001), although the difference in yield between the second and third samples was not significant. CONCLUSIONS: To enhance the yield of endocervical cells in Pap smear sampling, consideration should be given to using the cervical brush routinely as a sampling instrument and to taking more than one sample per screening.

Use of a heat exchanger to prevent hypothermia during thoracic and thoracoabdominal aneurysm repairs.

Hypothermia is a common unplanned occurrence in many patients undergoing repair of thoracic and thoracoabdominal aneurysms. Many undesirable side effects of hypothermia have been documented, including decreased cardiac output, conduction abnormalities, and blood coagulopathies. We have developed a simple system that incorporates a Sci-Med Biotherm heat exchanger into our left heart bypass circuit. This provides us with the ability to actively rewarm the patient safely and efficiently. This study looks at 16 consecutive patients undergoing repair of thoracic or thoracoabdominal aneurysms. In the 9 patients in whom the heat exchanger was used, there were no adverse effects related to the heat exchanger. All patients had significantly higher temperatures at the conclusion of the procedure than the 7 patients in whom the heat exchanger was not used.

The effectiveness of cytologic evaluation of products of conception.

In therapeutic abortions, the major issue of concern to physicians and patients is the documentation of an intrauterine pregnancy and its successful termination. We describe an inexpensive cytologic approach by which results can be obtained in a few hours and a permanent method of documentation provided. Three hundred cases of therapeutic abortion were evaluated by the cytologic method, and 93.7% of cases were positive, ie, they demonstrated the presence of nucleated red blood cells and/or placental villi. By routine histologic examination, 96.3% of the cases were positive. In only 2% of cases was the cytologic finding negative and the histologic finding positive. If the histologic finding is also negative, we recommend, based on our data, that the patient be reevaluated.

Gastric linitis plastica with metastases to the colon: a mimic of Crohn's disease.

A 60-year-old woman had progressive lower abdominal pain, nonbloody diarrhea, and weight loss, followed by severe epigastric pain and dysphagia. Radiographic evaluation of the colon showed segmental strictures which were interpreted as Crohn's disease. Medical treatment was not helpful. Neither gross endoscopic appearance nor multiple biopsies of the esophagus, stomach, and colon were diagnostic. Finally, laparotomy with full-thickness biopsies of the stomach and colon revealed linitis plastica. The clinician should be alert to colonic metastases from gastric linitis plastica, for it can produce focal or segmental strictures, mimicking more common colonic diseases such as Crohn's disease. A full-thickness biopsy is often necessary for a firm diagnosis. We review the literature on this occurrence, highlighting the clinical and radiologic spectrum, as well as the organ systems most often affected when gastric linitis plastica metastasizes.