Display of three-parametric data acquired by a flow cytometer

A program for producing contour maps of three-parameter flow cytometric data is described. The program is written in Pascal and can be implemented on any microcomputer with a graphics screen.     

MHC mismatch inhibits neurogenesis and neuron maturation in stem cell allografts

Background

The role of histocompatibility and immune recognition in stem cell transplant therapy has been controversial, with many reports arguing that undifferentiated stem cells are protected from immune recognition due to the absence of major histocompatibility complex (MHC) markers. This argument is even more persuasive in transplantation into the central nervous system (CNS) where the graft rejection response is minimal.

Methodology/Principal Findings

In this study, we evaluate graft survival and neuron production in perfectly matched vs. strongly mismatched neural stem cells transplanted into the hippocampus in mice. Although allogeneic cells survive, we observe that MHC-mismatch decreases surviving cell numbers and strongly inhibits the differentiation and retention of graft-derived as well as endogenously produced new neurons. Immune suppression with cyclosporine-A did not improve outcome but non-steroidal anti-inflammatory drugs, indomethacin or rosiglitazone, were able to restore allogeneic neuron production, integration and retention to the level of syngeneic grafts.

Conclusions/Significance

These results suggest an important but unsuspected role for innate, rather than adaptive, immunity in the survival and function of MHC-mismatched cellular grafts in the CNS.     

Nitric oxide function in the skin.

Endogenously produced nitric oxide (NO) has a remarkably diverse range of biological functions, including a role in neurotransmission, smooth muscle relaxation, and the response to immunogens. Over the last 10 years, it has become clear that this extraordinary molecular messenger also plays a vital role in the skin, orchestrating normal regulatory processes and underlying some of the pathophysiological ones. We thought it pertinent to review the current literature concerning the possible function of NO in normal skin, its clinical and pathological significance, and the potential for therapeutic advances. The keratinocytes, which make up the bulk of the epidermis, constitutively express the neuronal isoform of NO synthase (NOS1), whereas the fibroblasts in the dermis and other cell types in the skin express the endothelial isoform (NOS3). Under certain conditions, virtually all skin cells appear to be capable of expressing the inducible NOS isoform (NOS2). The expression of NOS2 is also strongly implicated in psoriasis and other inflammatory skin conditions. Constitutive, low level NO production in the skin seems to play a role in the maintenance of barrier function and in determining blood flow rate in the microvasculature. Higher levels of NOS activity, stimulated by ultraviolet (UV) light or skin wounding, initiate other more complex reactions that require the orchestration of various cell types in a variety of spatially and temporally coordinated sets of responses. The NO liberated following UV irradiation plays a significant role in initiating melanogenesis, erythema, and immunosuppression. New evidence suggests that it may also be involved in protecting the keratinocytes against UV-induced apoptosis. The enhanced NOS activity in skin wounding (reviewed recently in this journal [Nitric oxide 7 (2002) 1]) appears to be important in guiding the infiltrating white blood cells and initiating the inflammation. In response to both insults, UV irradiation and skin wounding, the activation of constitutive NOS proceeds and overlaps with the expression of NOS2. Thus, at a macro-level, at least three different rates of NO production can occur in the skin, which seem to play an important part in organizing the skin's unique adaptability and function.     

The destruction of allogeneic tumour cells by antibody and adherent cells from peritoneal cavities of mice.

Antisera to a DBA2 lymphoma (SL2) were raised in C57 black mice. The sera contained cell-dependent antibodies which lysed SL2 cells in conjunction with a monolayer of adherent peritoneal cells from unimmunised mice. The strongest lytic reaction was observed when the three components of the system, monolayer, target, and antiserum, were incubated together. The free antibody was not cytophilic for macrophages. It combined specifically with the target cell but precoated SL2 cells were not lysed effectively, probably because cells in the monolayer also accelerated the inactivation of antibody on the surface of the target cell.     

Long-term effects of catchment liming on invertebrates in upland streams

1. Catchment liming to mitigate acidification causes major chemical change in freshwaters but longer‐term effects are poorly understood. Using a replicated basin‐scale experiment with a multiple BACI design (= before‐after‐control‐impact), we assessed chemical and biological effects for 10 years after the catchments of three acidified Welsh streams at Llyn Brianne were limed in 1987/88. 2. Stream chemistry was measured weekly to monthly, and macroinvertebrates monitored annually, between 1985 and 1998. Biological change through time was assessed from the abundance and taxon richness of invertebrates. We paid particular attention to 18 species known to be acid‐sensitive. The effects of liming were assessed by comparing chemical and biological trends among the three replicate limed streams, three acid reference streams and two naturally circumneutral streams. 3. Following single lime applications, acid‐base chemistry in treated streams changed significantly. High mean pH (> 6), increased calcium (> 2.5 mg L^?1) and low aluminium (< 0.1 mg L^?1) persisted throughout the 10 years following liming. 4. The effects of liming on invertebrates were modest. Acid sensitive taxa increased significantly in abundance in limed streams, but only during 2 years following treatment. Significant effects on richness were more sustained, but on average added only 2–3 acid‐sensitive species to the treated streams, roughly one‐third of their average richness in adjacent circumneutral streams. Only the mayfly Baetis rhodani and the stonefly Brachyptera risi occurred significantly more often in limed streams after treatment than before it. 5. Despite these modest long‐term effects on invertebrates, nearly 80% of the total pool of acid‐sensitive species has occurred at least once in the limed streams in the 10 years since treatment. This pattern of occurrence suggests that the colonization of limed streams by acid‐sensitive taxa reflects limited persistence rather than restricted dispersal. We present evidence to show that episodes of low pH continued to affect acid‐sensitive taxa even after liming. We highlight the importance of extending the time‐periods over which the effects of large‐scale ecological experiments are assessed.