Optical response of the cultured bovine lens; testing opaque or partially transparent semi-solid/solid common consumer hygiene products

This study determines the relative ocular lens irritancy of 16 common partially transparent or non-transparent consumer hygiene products. The irritancy was found by measuring the changes in the sharpness of focus [referred to as the back vertex distance (BVD) variability] of the cultured bovine lens using a scanning laser In Vitro Assay System. This method consists of a laser beam that scans across the lens, and a computer, which then analyses the average focal length (mm), the BVD variability (mm), and the intensity of the beam transmitted. Lenses were exposed to the 16 hygiene products and the lens’ focusing ability was monitored over 192 h. The products are semi-solids or solids (e.g. gels, lotions, shampoos). They are categorized into six groups: shampoos, body washes, lotions, toothpastes, deodorant, and anti-perspirant. Damage (measured by > 1 mm BVD variability) occurred slower for the shampoos, especially in the case of baby shampoo. The results indicate that shampoos exhibit the lowest level of ocular lens toxicity (irritability) while the deodorant is the most damaging.     

DIAGNOSIS OF CONGENITAL CYTOMEGALOVIRUS INFECTION BY EXAMINATION OF PLACENTA: Application of Polymerase Chain Reaction and in Situ Hybridization

We examined the placentas of 12 patients in whom congenital cytomegalovirus CMV infection was suspected from serological and or pathological evaluation. Seven patients died including four intrauterine deaths and five survived. On histological examination, the characteristic inclusion bodies were detected in only three placentas, and villitis with plasma cell infiltration was seen in eight placentas. Immunohistochemistry using a specific antibody against CMV improved the sensitivity of CMV detection 10 cases were positive . With the polymerase chain reaction PCR following the extraction of DNA from formaldehyde-fixed placenta samples, CMV DNA was detected in seven cases. All 12 subjects were diagnosed with CMV infection by additional Southern blot analysis after the PCR. CMV DNA was also detected by an in situ hybridization method in all cases. With current molecular biological techniques the placenta can be reliably used for the diagnosis of congenital CMV infection.     

In vitro T-lymphocyte function in head and neck cancer patients

T-lymphocyte cell function was studied in vitro in peripheral blood mononuclear cells (PBMC) from 61 male patients with head and neck squamous cell carcinomas compared to 46 control patients. Patients older than 80 years or with reduced tumor-related performance status as measured by Karnofsky score less than 75 were excluded. In contrast to previous similar studies, control subjects ensured a minimum stress load by sampling all patients on the day of either diagnostic or therapeutic surgery. PBMC were separated by density-gradient centrifugation and subsequently cultured with autologous sera in vitro. The mitogen concanavalin A (Con A), which stimulates all T-cell clones, was employed. Findings showed that increased Con A stimulation and PBMC proliferation occurred with PBMC from cancer patients compared to that from control patients. In contrast, no differences could be detected with respect to the stimulated supernatant level of interleukin-2, interleukin-4 or interferon-γ between the groups. These results suggest that T-lymphocytes from PBMC are generally affected by neoplastic disease through either a supporting cell or serum factor.     

The change in tenascin expression in mouse uterus during early pregnancy

Purpose: Our aim was to examine the changes in spatiotemporal tenascin (TN) expression in mouse uterus during early pregnancy, when the uterine tissue undergoes a tremendous restructuring. Methods: Using immunohistochemistry and in situ hybridization, the changes in distribution of TN protein in mouse uterine tissues in pregnancy Day 0 through Day 5 were analyzed. Results: Immunoreactive TN and TN mRNA were expressed in the basement membrane of the epithelium as well as in the smooth muscle layer, and their distribution shifted from the subbasement region on Day 0–3 to the smooth muscle layer on Days 4 and 5. Conclusions: These results indicate that TN expression in the uterus during early pregnancy is spatiotemporally different and may be regulated by a different mechanism.     

Evaluating sex chromosome content of sorted human sperm samples with use of dual-color fluorescence in situ hybridization

OBJECTIVE: Although most methods for selecting the sex of offspring by sorting spermatozoa are ineffective at shifting the ratio of Y- to X-containing cells, some commercial sources continue to offer such services. Our objective was to evaluate commercially “sorted” samples with use of dual-color fluorescence in situ hybridization and to identify variations in assessment by comparing motile and total sperm populations, donors, observers, and fluorescence in situ hybridization probes.STUDY DESIGN: Cryopreserved sperm from seven anonymous donors were processed as for insemination. Sperm cells from each total sample or motile subfraction were prepared for fluorescence in situ hybridization by incubation with disulfide-reducing agents to expand sperm nuclei. Two sets of X and Y chromosome–specific, fluorophore-labeled deoxyribonucleic acid probes were used. At least 400 nuclei from each preparation were classified independently by three blinded observers. Hybridization efficiency, aneuploidy, and sex chromosome content were evaluated in subsets of five unsorted, five female-oriented, and five male-oriented samples. Total and motile subfractions were compared with eight samples. Fluorescence in situ hybridization probes were compared in five paired unsorted samples.RESULTS: No differences were detected between washed samples and paired motile subfractions. No differences in hybridization and aneuploidy were detected between groups of sorted samples. The Y/X ratio was significantly different between the sorted groups. However, male-oriented samples had a lower Y/X ratio than female-oriented samples did. Observer and probe choice accounted for small but significant variations that did not alter conclusions about the Y/X ratio for sorted samples.CONCLUSION: In a series of 10 sorted samples from one commercial source, dual-color fluorescence in situ hybridization demonstrated a small but significant shift in the sex chromosome ratios among samples. However, this shift was opposite to that expected by the orientation of the sorted samples. (Am J Obstet Gynecol 1997;176:1172-80.)     

MYCN AMPLIFICATION IN MEDULLOBLASTOMA

Prominent neuronal differentiation and MYCN amplification occur only in a small percentage of medulloblastomas (primitive neuroectodermal tumor of the cerebellum). In this article, we describe two medulloblastomas that showed a striking degree of neuronal differentiation with islands of mature neurons associated with abundant neuropil. In these differentiated foci, there were no mitoses or atypia, and the histology was reminiscent of a malformative or hamartomatous lesion. Both tumors were assessed for MYCN amplification by the novel technique of differential polymerase chain reaction (PCR) and fluorescent in situ hybridization (FISH). In case 1, MYCN amplification was in the 10 - fold range determined by differential PCR, while in case 2 the level of amplification was marked, with 20 - 30 copies compared with diploid controls. FISH analysis also confirmed the presence of MYCN amplification in both tumors. These two cases are of great interest, especially as they show MYCN amplification in medulloblastomas with marked degree of neuronal differentiation. Furthermore, in both cases, there was evidence of a slower and more indolent clinical course, suggesting a more favorable outcome in medulloblastoma with this degree of neuronal differentiation.     

The effect of interleukin-6 on enhancing the invasiveness of head and neck cancer cells in vitro

Patients with head and neck cancers that produce a high concentration of granulocyte colony-stimulating factor (G-CSF) or patients with esophageal squamous cell carcinomas who have elevated serum interleukin-6 (IL-6) concentrations have been found previously to be at significant risk for tumor invasion to adjacent organs as well as frequent metastases. This suggests that G-CSF and Il-6 enhance the invasiveness and metastatic potential of cancer cells. We studied the in vitro invasiveness of head and neck cancer cell lines with and without recombinant human G-CSF (rhG-CSF) and human IL-6 (hIL-6) in an extracellular matrix membrane system. The degree of invasiveness was affected by incubating cells with hIL-6, but not by pre-incubating the cell-matrix with hIL-6. The maximum concentration of hIL-6 for enhanced invasiveness was approximately 5,000 u/ml. In addition, rhG-CSF enhanced the invasiveness of tumor cells that produced large amounts of G-CSF. The present study also suggests that tumor cells tend to invade and metastasize in an environment rich in hIL-6. Received: 3 November 1997 / Accepted: 1 April 1998     

Retrospective analysis of clonality and detection of residual disease in myeloid leukemia by FISH on long-term stored bone marrow smears

Abstract Background. Fluorescence in situ hybridization (FISH) has allowed the detection of numerical chromosomal aberrations in interphase nuclei on fresh or frozen smears of leukemia.
Methods. To analyze clonality and residual disease in myeloid leukemia retrospectively, we applied FISH to bone marrow smears stored at ambient temperature for up to 9 years.
Results: When hybridization efficiency was investigated on stored control smears from patients without hematological malignancy, more than 96% of nuclei showed the expected number of signals using DNA probes specific for chromosome 7, X or Y. In combination with cell morphology, we observed much higher hybridization efficiency in blasts and granulomonocytic cells compared with lymphoid and erythroid cells. On the basis of good hybridization efficiency for old smear specimens, we applied FISH to stored bone marrow smears of myeloid leukemias, in which either loss of chromosome 7 or loss of sex chromosomes had been verified previously by conventional cytogenetics (one patient with chronic myelomonocytic leukemia (CMML) and four with acute myeloid leukemia (AML; three M2 and one M7)). As a result, the loss of chromosome was detected in blasts from all patients and was observed in mature granulocytes, except in M7. In the CMML patient and one AML (M2) patient with t(8;21), lymphoid and erythroid cells also showed the loss of chromosomes, suggesting that it should occur at stem-cell level. A high amount of residual disease was detected in the morphological remission samples in one AML (M2) patient after induction therapy. The patient eventually succumbed to relapse.
Conclusion Thus, the present FISH technique is useful to analyze the clinical significance of clonality and the residual disease in myeloid leukemia, retrospectively.     

In vivo neutron dosimetry during high-energy Bremsstrahlung radiotherapy

Purpose: A new technique is presented for in vivo measurements of the dose equivalent from photoneutrons produced by high-energy radiotherapy accelerators.Methods and Materials: The dosimeters used for this purpose are vials of superheated halocarbon droplets suspended in a tissue-equivalent gel. Neutron interactions nucleate the formation of bubbles, which can be recorded through the volume of gel they displace from the detector vials into graduated pipettes. These detectors offer inherent photon discrimination, dose-equivalent response to neutrons, passive operation, and small sensitive size. An in vivo vaginal probe was fabricated containing one of these neutron detector vials and a photon-sensitive diode. Measurements were carried out in patients undergoing high-energy x-ray radiotherapy and were also repeated in-phantom, under similar irradiation geometries.Results and Conclusion: Neutron doses of 0.02 Sv were measured in correspondence to the cervix, 50 cm from the photon beam axis, following a complete treatment course of 46.5 Gy with an upper mantle field of 18-MV x-rays. This fraction of dose from neutrons is measured reliably within an intense photon background, making the technique a valid solution to challenging dosimetry problems such as the determination of fetal exposure in radiotherapy. These measurements can be easily carried out with tissue-equivalent phantoms, as our results indicate an excellent correlation between in vivo and in-phantom dosimetry.     

体外受精-胚胎移植治疗不孕症85例报道

目的：评价应用体外受精－胚胎移植（ＩＶＦ－ＥＴ）治疗８５例不孕患者的临床结果。方法：用药方案有：①ＧｎＲＨａ／ＦＳＨ／ｈＭＧ；②ＧｎＲＨａ／ＦＳＨ；③ＧｎＲＨａ／ｈＭＧ。培养液包括Ｅａｒｌｅ’ｓ平衡盐液及人类输卵管液（ＨＴＦ）。结果：８５例进行了１０９个ＩＶＦ周期治疗，１２个周期（１１０％）因反应不良而取消。每周期获取卵母细胞（１０２±５４）个，卵子受精率６８９％，卵裂率６０８％，每个转移胚胎植入率７７％（３１／４０２）。９４个移植周期中，每周期移植（４９±２４）个胚胎，获２０次临床妊娠（２１３％）。采用ＨＴＦ妊娠率为２２４％，高于应用Ｅａｒｌｅ’ｓ液的妊娠率（１８５％），但差异无统计学意义。２０次妊娠患者中，１次输卵管妊娠，４次自然流产（２０％），５例足月分娩，１０例继续妊娠。多胎妊娠率４０％，重度ＯＨＳＳ发生率４１％。结论：ＩＶＦ－ＥＴ是治疗输卵管因素、子宫内膜异位症等不孕症的重要而有效的手段。体外培养时用ＨＴＦ可能优于Ｅａｒｌｅ’ｓ液，而有利于提高妊娠率。