Cryohemolysis for the detection of hereditary spherocytosis: Correlation studies with osmotic fragility and autohemolysis

Laboratory methods aimed to assess the presence of spheroidal cells such as osmotic fragility, autohemolysis, and glycerol lysis time are very elaborate, time consuming, and often give inconclusive results. We have developed a diagnostic test based on a unique sensitivity of HS cells to hypertonic cryohemolysis and analyzed blood samples of 55 HS patients. The patients were divided into two subgroups, clinically affected probands and their relatives. To get quantitative comparisons with the classic methods, the cryohemolysis results were compared to two parameters of the osmotic fragility test: the salt concentration that causes 50% hemolysis, and the percent lysis at a constant salt concentration. Autohemolysis results were also compared. To evaluate which of these tests has the best analytical power, we calculated the mean results and 2 SDs of each parameter in a control group, and then looked to see which of them was best in identifying the patients. The cryohemolysis test was the single parameter that identified all cases including asymptomatic carriers of the disease. The ability of this test to identify the less severe cases probably reflects the dependency of the cryohemolysis on factors that are more related to the primary membrane molecular defects and less by the surface area to volume ratio. Am. J. Hematol. 58:206–212, 1998. © 1998 Wiley-Liss, Inc.     

The role of a human chorionic gonadotropin burst in in vitro fertilization

Findings: No oocytes were found during four ovum pickups (OPU), despite a satisfactory ovarian response to controlled ovarian hyperstimulation. After the first attempt failed in the fourth case, five eggs were retrieved, fertilized, and cleaved after cycle rescue with hCG. Conclusions: Whenever oocytes are not aspirated during OPU due to a lack of hCG administration, the cycle may be rescued if 10,000 IU of hCG is injected immediately and OPU planned for 33–36 hr later.     

Monoallelic gene expression: a repertoire of recurrent themes

Summary:  The development of mature B and T cells in the lymphoid system involves a series of molecular decisions that culminate in the expression of a single antigen receptor on the cell surface, a phenomenon termed allelic exclusion. While feedback inhibition of the recombinase-activation gene proteins evidently plays an important role in the maintenance of allelic exclusion, the initial restriction of rearrangement to only one allele in each cell seems to be achieved through monoallelic epigenetic changes. Epigenetic mechanisms involved in the establishment of allelic exclusion also play a central role in other types of monoallelic expression, including X-chromosome inactivation in female cells, and parental imprinting. In all three systems, the inequality of the two alleles seems to be achieved mainly by differential DNA methylation, asynchronous DNA replication, differential chromatin modifications, unequal nuclear localization, and non-coding RNA. In this review, we discuss the unifying features among these monoallelically expressed systems and the unique characteristics displayed by each of them.     

A comparison of trends in incidence and mortality rates of breast cancer, incidence to mortality ratio and stage at diagnosis between Arab and Jewish women in Israel, 1979-2002.

In Israel, breast cancer is the most common malignancy in women, but there are large intra-population differences. The aim of this study was to compare the incidence and mortality, incidence to mortality rate ratio and stage at diagnosis of breast cancer between Arab and Jewish women in Israel. Data on all cases of breast cancer, stage at diagnosis and mortality were obtained from the National Cancer Registry and the Central Bureau of Statistics. Trends in age-specific and age-adjusted incidence and mortality rates, rate ratios and stage at diagnosis were examined for Arab and Jewish women during 1979-2002. Five-year survival rates for 1995-1999 were compared by stage. Among Arab women, age-adjusted incidence rates increased by 202.1%, from 14.1 per 100,000 in 1979-1981 to 42.6 in 2000-2002. Among Jewish women, the rates increased by 45.7%, from 71.1 per 100,000 women in 1979-1981 to 103.6 in 2000-2002. Incidence to mortality rate ratio increased for both population groups, but it is still lower among Arab women. In every age group, Arab women were more likely to be diagnosed at a more advanced stage of the disease. The rise in breast cancer incidence and mortality rates and the later stage of diagnosis among Arab women emphasize the urgent need for increasing early detection of breast cancer in the Arab population by improving rates of compliance with screening mammography.     

Coming together in pain and joy: A multicultural and arts-based suicide awareness project

The issue of suicide is still taboo in many cultures and the silence surrounding this problem is often destructive. Those affected by suicide have few opportunities to express their pain and their communities are left feeling helpless. The creative arts therapies have been used to address the emotions surrounding suicide, however few projects have studied the use of the arts to promote suicide awareness. Furthermore, there is a lack of multicultural research in this area. This article describes a two-day arts-based symposium that brought together members from diverse cultural communities including Inuit, Mohawk, Jewish, Christian, Baha’i, South-Asian Canadian, Senior and LGBTQ (Lesbian, Gay, Bisexual, Trans-gendered, Queer) communities. Through drama, music, art-making and writing, participants explored this difficult subject, expressed complex feelings and shared their perspectives. An instrumental case study design was employed for this research project. The objectives were to gather information on participants’ experience of exploring the issue of suicide within an arts-based approach and to determine if cross-cultural themes would emerge. The findings indicate that using the arts helped to facilitate dialogue and communication and specific cross-cultural themes did emerge.     

Serum Soluble Triggering Receptor on Myeloid Cells-1 (sTREM-1) is Elevated in Systemic Lupus Erythematosus but does not Distinguish Between Lupus Alone and Concurrent Infection

We sought to determine serum triggering receptor expressed on myeloid cell-1 (sTREM-1) level in a cohort of patients with systemic lupus erythematosus (SLE). Serum sTREM-1 level of 98 patients with SLE and 49 healthy controls was assayed by ELISA. Serum sTREM-1 level was significantly elevated in a cohort of 78 unselected consecutively recruited patients with SLE (mean 1.1?±?2.8 ρg/ml, median 0.02 ρg/ml) compared to that of the controls (mean 0.11?±?0.3 ρg/ml, median 0 ρg/ml; p?<?0.0001). We also determined serum sTREM-1 level of 20 SLE patients with a concurrent infection (mean 0.6?±?1.1 ρg/ml, median 0.12 ρg/ml) and found it not statistically significant compared with that of the patients without infection. Serum sTREM-1 level did not correlate with SLE disease activity. Our finding of elevated serum sTREM-1 level suggests an increased shedding of TREM-1 in SLE and a possible novel pathway of innate immune response in autoimmunity.     

Monocyte chemoattractant protein-1 in the aqueous humour of patients with age-related macular degeneration

Background: To investigate the role of inflammation in age‐related macular degeneration by measuring the levels of cytokines in the aqueous humour. Methods: Samples of aqueous humour were collected from 34 patients with age‐related macular degeneration and 16 age‐matched control subjects undergoing cataract surgery. Age‐related macular degeneration stage was determined clinically, before surgery. Levels of cytokines were measured using Luminex X‐MAP technology, and positive results were verified by Western blot. Results: Age‐related macular degeneration was moderate in 18 patients and advanced in 16. The advanced age‐related macular degeneration group was further divided into patients with active choroidal neovascularization (n = 7), disciform scar (n = 7) or central geographic atrophy (n = 2). Higher‐than‐normal levels of monocyte chemoattractant protein‐1 in the aqueous humour were associated with advanced age‐related macular degeneration (200 ± 140 pg/mL vs. 100 ± 61 pg/mL; P = 0.03), especially active choroidal neovascularization (255 ± 155 pg/mL; P = 0.02), Western blot analysis verified the monocyte chemoattractant protein‐1 findings. Patients with disciform scar showed a trend of abnormally high levels of interleukin‐12 (p70) (1.7 ± 2.4 pg/mL vs. 0.2 ± 1 pg/mL; P = 0.07), tumour necrosis factor‐α (1.8 ± 2.4 pg/mL vs. 0.3 ± 1 pg/mL; P = 0.06) and interleukin‐12 (4.7 ± 6.4 pg/mL vs. 1.2 ± 2.1 pg/mL; P = 0.08). Conclusion: Elevated levels of inflammation‐related cytokines in the aqueous humour in various stages of age‐related macular degeneration may suggest a pathogenic role of inflammation. Monocyte chemoattractant protein‐1 may be indicative of the angiogenic phase. Further corroborative studies are required.     

Rapid detection of blaKPC carbapenemase genes by internally controlled real-time PCR assay using bactec blood culture bottles

Rapid detection of drug-resistant bacteria in clinical samples plays an instrumental role in patients' infection management and in implementing effective infection control policies. In the study described in this report, we validated a multiplex TaqMan real-time quantitative PCR (qPCR) assay for the detection of bla(KPC) genes and the human RNase P gene in Bactec blood culture bottles. The MagNA Pure LC (version 2.0) instrument was utilized to extract nucleic acids from the inoculated broth, while bovine serum albumin (BSA) was utilized as the PCR inhibitor reliever. The multiplex assay, which was specific for the detection of bla(KPC) genes, had a limit of detection of 19 CFU per reaction mixture with human blood-spiked Bactec bottles. Of the 323 Bactec blood culture sets evaluated, the same 55 (17%) blood cultures positive for carbapenem-resistant bacteria by culture were also positive by the validated qPCR assay. Thus, the sensitivity, specificity, positive predictive value, and negative predictive value of the qPCR assay compared to the results of culture were all 100%. bla(KPC) genes were also detected from the same Bactec bottle broth after manual extraction with a QIAamp DNA minikit; however, there was an average 3-threshold-cycle delay in the qPCR readings. With the limited therapeutic options available, the accurate and rapid detection of bla(KPC)-possessing bacteria by the described bla(KPC)/RNase P assay will be a crucial first step in ensuring optimal clinical outcomes and infection control.