An improved method for preparing refrozen rethawed human lymphocytes on plates for microcytotoxicity studies.

This report describes simplified methods for the initial freezing and thawing of human lymphocytes and the subsequent use of these cells after refreezing on cytotoxicity plates, storage, and a second thaw. The proposed initial freeze method eliminates some technical inconveniences required previously such as chilling of cells prior to addition of DMSO, preparing cryoprotective mixtures just prior to freezing, controlled rate of freezing and thawing and the washing of cells after thawing. However, pH of the media, blood freshness, type of storage tube used, and constant storage temperature were found to be very important to maintain good cell viability. Most lymphocytes maintain an average viability of 85 to 95% for at least a year when prepared according to the present freezing and thawing technique.When panels of lymphocytes are prepared for refrozen rethawed cytotoxicity test plates, the thaw time between freezes must be brief. Production of test plates on ice, however, was not found to be necessary. As the period of storage of refrozen cells on plates increases, viability of the cells after a second thaw decreases and treatment with DNase to enzymatically remove the dead cells is useful. With this procedure, refrozen rethawed lymphocytes up to a year old can be prepared on microcytotoxicity test plates with average viabilities of 90 ± 1%.     

Cover Image: Plasmodium berghei sporozoites in nonreplicative vacuole are eliminated by a PI3P‐mediated autophagy‐independent pathway (Cellular Microbiology 1/2021)

Immunofluorescence image of Plasmodium berghei sporozoites (red) in HeLa cells 1.5 hours post infection. The parasitophorous vacuole membrane (PVM) that surrounds the parasites was stained with antibodies against the PVM‐resident protein UIS4 (grey). DNA was stained with DAPI (blue). Imaging was performed on a Leica SP8 confocal microscope. For further details, readers are referred to the article by Bindschedler et al. on p. e13271 of this issue.

     

Cardiac magnetic resonance imaging in daily practice in a peripheral medical centre: description of the first 383 patients

Background. Cardiac magnetic resonance (CMR) imaging has evolved over the last decade into an indispensable diagnostic instrument. CMR imaging noninvasively provides structural, functional and morphological information with high spatial resolution and an unlimited field of view. Since October 2006 the VieCuri Medical Centre in Venlo has a CMR scanner at its disposal.Objectives. The goal of this study was to analyse the impact of CMR imaging on diagnosis and treatment in daily practice in the setting of a medium-volume peripheral hospital.Methods. All patients who underwent CMR imaging between October 2006 and November 2008 were included in this analysis. The medical history before and after the CMR scan, the application form for CMR imaging and the outcome of the scans were reviewed. CMR images, obtained using a 1.5-T magnetic resonance imaging system, were reviewed by a multidisciplinary team.Results. In 235 patients CMR imaging demonstrated one or more abnormalities, whereas CMR imaging did not identify any abnormalities in 148 patients. CMR imaging confirmed an expected finding in 166 cases, identified an unexpected condition in 69 cases, ruled out an expected finding in 59 cases and ruled out a suspected condition in 89 cases. Due to better insight into diagnosis, CMR imaging resulted in a change of treatment in 166 of the total of 383 CMR scans (43%).Conclusion. In a relevant number of cases CMR imaging leads to a change in the treatment of a patient, proving the value of CMR imaging as a diagnostic modality. Therefore, CMR imaging is an excellent opportunity for peripheral medical centres to improve efficiency and the standard of patient care. (Neth Heart J 2010;18:524–30.)     

Vital rates of two small populations of brown bears in Canada and range‐wide relationship between population size and trend

Identifying mechanisms of population change is fundamental for conserving small and declining populations and determining effective management strategies. Few studies, however, have measured the demographic components of population change for small populations of mammals (<50 individuals). We estimated vital rates and trends in two adjacent but genetically distinct, threatened brown bear (Ursus arctos) populations in British Columbia, Canada, following the cessation of hunting. One population had approximately 45 resident bears but had some genetic and geographic connectivity to neighboring populations, while the other population had <25 individuals and was isolated. We estimated population‐specific vital rates by monitoring survival and reproduction of telemetered female bears and their dependent offspring from 2005 to 2018. In the larger, connected population, independent female survival was 1.00 (95% CI: 0.96–1.00) and the survival of cubs in their first year was 0.85 (95% CI: 0.62–0.95). In the smaller, isolated population, independent female survival was 0.81 (95% CI: 0.64–0.93) and first‐year cub survival was 0.33 (95% CI: 0.11–0.67). Reproductive rates did not differ between populations. The large differences in age‐specific survival estimates resulted in a projected population increase in the larger population (λ = 1.09; 95% CI: 1.04–1.13) and population decrease in the smaller population (λ = 0.84; 95% CI: 0.72–0.95). Low female survival in the smaller population was the result of both continued human‐caused mortality and an unusually high rate of natural mortality. Low cub survival may have been due to inbreeding and the loss of genetic diversity common in small populations, or to limited resources. In a systematic literature review, we compared our population trend estimates with those reported for other small populations (<300 individuals) of brown bears. Results suggest that once brown bear populations become small and isolated, populations rarely increase and, even with intensive management, recovery remains challenging.     

“We Went Out to Explore,But Gained Nothing But Illness”: Immigration Expectations,Reality, Risk and Resilience in Chinese-Canadian Women with a History of Suicide-Related Behaviour

Suicide is a complex and tragic outcome driven by biological, psychological, social and cultural factors. Women of Chinese descent and women who have immigrated to other countries have higher rates of suicidal ideation and behaviour, and immigration-related stress may contribute. To understand the experiences of immigration and their relationship with distress and suicide-related behaviour in Chinese women who have immigrated to Canada. 10 semi-structured qualitative interviews with Chinese women who have immigrated to Toronto, Canada and have a history of suicide-related behaviour were completed and analyzed using a constructivist grounded theory methodology. Immigration-related and acculturation stress stemmed from unmet expectations and harsh realities. These repeated experiences resulted in hopelessness, helplessness, and alienation, which are risk factors for suicide and suicide-related behaviour. However, immigration-related support can also increase hope, self-efficacy and connectedness to foster recovery and resilience. This is the first qualitative study focusing on immigration experiences and its relationship to suicide-related behaviour in Chinese immigrant women. Knowledge of immigration and acculturation stressors can a) help identify and support women at risk for suicide and b) form a target for social intervention for all immigrant women, regardless of suicide risk.