Physical and chemical limnology of alpine lakes and pools in the Rwenzori Mountains (Uganda–DR Congo)

This study describes the physical and chemical properties of 17 Afroalpine lakes (>2 m deep) and 11 pools (<2 m deep) in the Rwenzori mountains, Uganda-DR Congo, with the aim to establish the baseline conditions against which to evaluate future environmental and biological changes in these unique tropical ecosystems, and to provide the foundation for lake-based paleoenvironmental studies. Most Rwenzori lakes are located above 3,500 m elevation, and dilute (5–52 μS/cm specific conductance at 25°C) open systems with surface in- and outflow. Multivariate ordination and pairwise correlations between environmental variables mainly differentiate between (1) lakes located near or above 4,000 m (3,890–4,487 m), with at least some direct input of glacial meltwater and surrounded by rocky catchments or alpine vegetation; and (2) lakes located mostly below 4,000 m (2,990–4,054 m), remote from glaciers and surrounded by Ericaceous vegetation and/or bogs. The former group are mildly acidic to neutral clear-water lakes (surface pH: 5.80–7.82; Secchi depth: 120–280 cm) with often above-average dissolved ion concentrations (18–52 μS/cm). These lakes are (ultra-) oligotrophic to mesotrophic (TP: 3.1–12.4 μg/l; Chl-a: 0.3–10.9 μg/l) and phosphorus-limited (mass TN/TP: 22.9–81.4). The latter group are mildly to strongly acidic (pH: 4.30–6.69) waters stained by dissolved organic carbon (DOC: 6.8–13.6 mg/l) and more modest transparency (Secchi-disk depth: 60–132 cm). Ratios of particulate carbon, particulate nitrogen and chlorophyll a in these lakes indicate that organic matter in suspension is primarily derived from the lakes’ catchments rather than aquatic primary productivity. Since key features in the Rwenzori lakes’ abiotic environment are strongly tied to temperature and catchment hydrology, these Afroalpine lake ecosystems can be expected to respond sensitively to climate change and glacier melting. Electronic supplementary material The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

Effects of cyclic nucleotide dependent protein kinases on the endoplasmic reticulum Ca2+ pump of bovine pulmonary artery

This paper describes the stimulation by cyclic nucleotide dependent protein kinases on the Ca2+ uptake by isolated endoplasmic reticulum (ER) vesicles from the bovine main pulmonary artery. This ER fraction has previously been shown to be highly enriched in phospholamban, a protein kinase substrate that has been well characterized in cardiac sarcoplasmic reticulum (SR), where its phosphorylation is accompanied by an increased rate of Ca2+ uptake. As previously observed for the phosphorylation of phospholamban, the stimulation of the rate of Ca uptake was as high with cGMP dependent protein kinase as with cAMP dependent protein kinase. The effect of phosphorylation of the ER membranes from smooth muscle on the Ca2+ uptake was smaller than that seen in cardiac SR, and it was only observed if albumin was included during the isolation of the membranes. This relatively small effect is probably not due to a lower ratio of phospholamban to Ca2(+)-transport enzyme in the ER membranes as compared to cardiac SR. Several alternative explanations are discussed.     

HDAC6 inhibition restores TDP‐43 pathology and axonal transport defects in human motor neurons with TARDBP mutations

TDP‐43 is the major component of pathological inclusions in most ALS patients and in up to 50% of patients with frontotemporal dementia (FTD). Heterozygous missense mutations in TARDBP, the gene encoding TDP‐43, are one of the common causes of familial ALS. In this study, we investigate TDP‐43 protein behavior in induced pluripotent stem cell (iPSC)‐derived motor neurons from three ALS patients with different TARDBP mutations, three healthy controls and an isogenic control. TARDPB mutations induce several TDP‐43 changes in spinal motor neurons, including cytoplasmic mislocalization and accumulation of insoluble TDP‐43, C‐terminal fragments, and phospho‐TDP‐43. By generating iPSC lines with allele‐specific tagging of TDP‐43, we find that mutant TDP‐43 initiates the observed disease phenotypes and has an altered interactome as indicated by mass spectrometry. Our findings also indicate that TDP‐43 proteinopathy results in a defect in mitochondrial transport. Lastly, we show that pharmacological inhibition of histone deacetylase 6 (HDAC6) restores the observed TDP‐43 pathologies and the axonal mitochondrial motility, suggesting that HDAC6 inhibition may be an interesting therapeutic target for neurodegenerative disorders linked to TDP‐43 pathology.     

Matrix effects break the LC behavior rule for analytes in LC-MS/MS analysis of biological samples

High-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are generally accepted as the preferred techniques for detecting and quantitating analytes of interest in biological matrices on the basis of the rule that one chemical compound yields one LC-peak with reliable retention time (Rt.). However, in the current study, we have found that under the same LC-MS conditions, the Rt. and shape of LC-peaks of bile acids in urine samples from animals fed dissimilar diets differed significantly among each other. To verify this matrix effect, 17 authentic bile acid standards were dissolved in pure methanol or in methanol containing extracts of urine from pigs consuming either breast milk or infant formula and analyzed by LC-MS/MS. The matrix components in urine from piglets fed formula significantly reduced the LC-peak Rt. and areas of bile acids. This is the first characterization of this matrix effect on Rt. in the literature. Moreover, the matrix effect resulted in an unexpected LC behavior: one single compound yielded two LC-peaks, which broke the rule of one LC-peak for one compound. The three bile acid standards which exhibited this unconventional LC behavior were chenodeoxycholic acid, deoxycholic acid, and glycocholic acid. One possible explanation for this effect is that some matrix components may have loosely bonded to analytes, which changed the time analytes were retained on a chromatography column and interfered with the ionization of analytes in the MS ion source to alter the peak area. This study indicates that a comprehensive understanding of matrix effects is needed towards improving the use of HPLC and LC-MS/MS techniques for qualitative and quantitative analyses of analytes in pharmacokinetics, proteomics/metabolomics, drug development, and sports drug testing, especially when LC-MS/MS data are analyzed by automation software where identification of an analyte is based on its exact molecular weight and Rt.     

Cerebro-oculo-facio-skeletal (COFS) syndrome. The variability of presenting symptoms as a manifestation of two subtypes?

In this paper we present two female newborns with the cerebro-oculo-facio-skeletal (COFS) syndrome. The variability of presenting symptoms and differences in clinical evolution of the two infants favour the suggestion of Preus that two subtypes of this autosomal recessive inherited syndrome may exist.     

Recruitment of inflammatory cells to a tumor deposit potentiates the immunotherapeutic effects of interleukin-2

Summary Interleukin-2 (IL-2) is a potent immunotherapeutic agent in murine models of intraperitoneal, pulmonary, and hepatic tumor implantation. Because of the systemic toxicity documented at doses of IL-2 required to control tumor growth, potentiation of the effects of low dose IL-2 is an important problem in immunotherapy. To address this problem, we attempted to recruit lymphocytes into a tumor mass. Allogeneic P185 (H-2^d) tumor was mixed with MCA-105 (H-2^b) tumor and injected s. c. into C57BL/6 (H-2^b) mice. Mice were treated with 50,000 units of IL-2 twice daily from day 0 to day 6. When IL-2 alone was used to treat s. c. tumor, there was no reduction in the size of tumor implants. When allogeneic tumor was mixed with syngeneic tumor, there was a reduction in tumor size at the high dose of allogeneic tumor but not at the low dose. When allogeneic tumor was mixed with syngeneic tumor and the mice treated with IL-2, the immunotherapeutic effects of IL-2 were markedly increased. These studies show that an immune response to alloantigens, generated within tumor tissue can augment the immunotherapeutic effects of exogenous IL-2.