Intracellular distribution and effect of the antimalarial drug mefloquine on lysosomes of rat liver

Abstract: Mefloquine was administered in a single dose (1–30 mg/100 g) to rats in order to study its subcellular distribution and effects on rat liver lysosomal structure and function. Subcellular fractionation showed a significant enrichment of mefloquine in lysosomes. Even repeated administration of mefloquine did not affect the levels of cytochrome-P-450 or its reductase, indicating, although not proving, that it is not metabolized by this mono-oxygenase system. Mefloquine caused an expansion of the lysosomal apparatus, earliest seen by 24 h and lasting for some 7 days. Initially, cytoplasmic constituents were seen inside the lysosomes. Later, the lysosomes harboured myelin-like figures (multilamellar bodies) disappearing after 7–10 days. The proteolytic and lipolytic capacity was assessed in isolated lysosomes. Mefloquine caused increased protein degradation but decreased breakdown of lipids. Concomitantly, all five major phospholipids (phosphatidyl-choline, -ethanolamine, -inositol, -serine and sphingomyelin) increased in the lysosomes. It is concluded that: (1) mefloquine is a lysosomotropic drug that accumulates in lysosomes; (2) mefloquine impairs lipid degradation with ensuing accumulation of lipids in lysosomes; and (3) lysosomal trapping explains the high volume distribution of mefloquine.     

Radiographic prediction of failure after fixation of cervical hip fracture

During 1984-1985, 410 patients with cervical hip fracture were randomized between 2 methods of internal fixation-a single nail (Rydell) or 2 LIH hook pins (LIH). The patients were followed-up prospectively for at least 2 years. Radiographs were taken after 1 week, 1, 3, 6, 12, and 24 months. The radiographs of the 295 paients alive 2 years postoperatively were examined by one of the authors. The sliding and the diversion of the pins and the nail in the anteroposterior projection and the diversion in the lateral projection were measured. In the failure group (non-union, late segmental collapse), the greatest sliding was noted within 1 month postoperatively and the diversion increased up to 3 months. Significant differences between the failure and the non-failure groups could be seen even after 1 week. We also found that the degree of sliding of the LIH pins and the Rydell nail 1 month postoperatively is comparable to the scintigraphic pattern 2 weeks postoperatively in predicting failure after nternal fixation of cervical hip fractures.     

Enhancement of porcine circovirus 2 replication in porcine cell lines by IFN-gamma before and after treatment and by IFN-alpha after treatment.

Stimulation of the porcine immune system causes increased replication of porcine circovirus 2 (PCV2) in vivo. In the present study, we investigated whether various cytokines (interleukin-1 [IL-1], IL-6, IL-10, tumor necrosis factor-alpha [TNF-alpha], interferon-alpha [IFN-alpha], and IFN-gamma) are able to influence PCV2 infection in vitro. No changes were observed in IL-1, IL-6, TNF-alpha, or IL-10-treated cells. However, it was demonstrated that IFN- alpha and IFN-gamma influenced PCV2 infection in porcine kidney cells (PK-15) and porcine monocytic cells (3D4/31). IFN-gamma added to the culture medium before, during, or after inoculation increased the number of PCV2 antigen- positive cells, respectively, by 418%, 171%, and 691% in PK-15 cells and by 706%, 114%, and 423% in 3D4/31 cells. IFN-alpha pretreatment decreased the number of infected PK-15 cells. When it was added after inoculation, IFN-alpha enhanced PCV2 infection by 529% in PK-15 cells and by 308% in 3D4/31 cells. The effect of both IFNs on PCV2 infection was dose dependent and could be blocked with IFN-alpha or IFN-gamma neutralizing antibodies. Leukocyte-derived porcine IFN-gamma induced a similar effect on PCV2 infection. Treatment of PK- 15 cultures with IFN-gamma caused a 20 times higher production of progeny virus. Confocal microscopy studies showed that the enhancing effect of IFN-gamma on PCV2 infection was achieved by increased internalization of PCV2 virionlike particles (VLPs). Binding of the VLPs to the cell or expression kinetics of PCV2 proteins in infected cells were not altered by IFN-gamma treatment. To our knowledge, this study reports the first enhancement of a viral infection by treatment with type I or type II IFNs.     

Impact of early dietary gamma-linolenic acid supplementation on atopic eczema in infancy

Polyunsaturated fatty acids (PUFAs) are components of cell membranes and may play an immunomodulating role in the pathogenesis of atopic dermatitis (AD). The goal was to determine the impact of PUFAs on AD by dietary supplementation of infants. Based on the parents' decision on their babies' primary feeding, mothers and newborns were randomized to the supplementation with gamma-linolenic acid (GLA) or placebo for up to 6 months. Breastfed infants received GLA by supplementing their mothers. Formula diet was commercial whey hydrolysate unsupplemented with PUFAs. Of 131 eligible infants, 24 developed AD within the first year of life. Of these, nine belonged to the exclusively breastfed group (n = 58), 14 to the combined-fed group (n = 53), and one to the never breastfed group (n = 20). We could not find an influence of GLA on the development of AD. In subjects with AD, at 1 yr of age the serum-immunoglobulin E (IgE) was the lowest in the GLA-supplemented group A-subjects. In the GLA-supplemented group, GLA-levels in breast milk were similar in atopic and non-atopic infants. In the non-supplemented group the GLA-content of breast milk was 0.07% of total fatty acids in atopic infants vs. 0.17% in non-atopic infants (p < 0.01). Dietary GLA-supplementation could not prevent AD. Interestingly, the number of infants developing AD was the lowest in never breastfed children. In infants suffering from AD, GLA-supplementation seemed to reduce total IgE in the first year of life.     

Platelet aggregation,disc haemorrhage and progressive loss of visual fields in glaucoma

Platelet aggregationin vitro, deterioration of visual field defects (VFD) and the prevalence of disc haemorrhages (DH) were assessed in 49 patients with primary open angle glaucoma (POAG) and compared with the findings for 67 individuals with suspected glaucoma (GS) in a seven-year follow-up study (range 5.8 to 8.2 years). The percentage patients with spontaneous platelet aggregation (SPA) was higher for POAG patients with visual field deterioration (60%) than both POAG patients without progressive loss of visual fields (12.5%; P<0.005) and those with suspected glaucoma (22.4%; P<0.005). The occurrence of DH was higher among POAG patients with progressive loss of visual field (28%) compared to the GS group (8.4%; P<0.025) and the group of patients consisting of POAG patients without deterioration of VFD and GS (9.9%; P<0.05). DH also occurred more often in patients with low tension glaucoma (41.6%) than in the remaining POAG patients (13.5%; P<0.05). No relation between the patients with SPA and the patients with DH was observed.Abbreviations NPB normal platelet behavior - SPA spontaneous platelet aggregation - DH disc haemorrhage     

Diagnosis of scaphoid fractures:A prospective multicenter study of 1,052 patients with 160 fractures

In a prospective multicenter study of 1,052 patients with clinical signs of a scaphoid fracture, mammo-graphic films and fine intensifying screens were used at the radiographic examination. 5 standardized projections including 3 special projections focused on the scaphoid were taken. 150 fractures were diagnosed at the first examination but in 10 cases the fracture was first diagnosed at a second radiographic examination after 10-14 days.

The second examination still seems mandatory despite the use of high quality radiographs with optimal spatial resolution and contrast, and the value of supplementary special projections.     

Identification of a 28 kDa lychee allergen as a triose-phosphate isomerase

The aim of this study was to investigate the allergenic potency of the exotic fruit lychee (Litchi chinensis SONN.). For this purpose the lychee protein extract was separated by ion exchange chromatography and a purified allergen with a molecular weight of 28 kDa was identified by N-terminal sequencing and peptide mass fingerprinting. Both methods determined the lychee allergen as a triose-phosphate isomerase. To this protein 67% of patients showed allergic reactions respectively IgE binding of their sera. Similar enzymes from other plants were also recognized as allergens.     

Light microscopic morphometric analysis of castration effects in the different lobes of the rat prostate

The morphologic effects of androgen deprivation in the different lobes of the rat prostate were examined by light microscopic morphometry. The prostates of Wistar male rats (260-340 g) were fixed in situ by glutaraldehyde perfusion in castrated animals 1 week after gonadectomy and in intact animals. The ventral (VP), dorsal (DP), and lateral (LP) lobes as well as the coagulating gland (CG) were dissected out, weighed, and processed for light microscopy. Using stereologic methods the following parameters were estimated for each lobe: volume fraction of connective tissue, epithelium and glandular lumina, average epithelial height, average epithelial cell volume, and total number of epithelial cells. Castration leads to a 58-76% reduction of the wet weight of all prostatic lobes. The decrease of glandular tissue is greater in VP than in LP, DP, and CG. In VP and LP, there is a 39-45% reduction of the epithelial height, and this effect is less pronounced in DP and CG. For all lobes, the shrinkage of average epithelial cell volume is in the same range (25-30%). Moreover, in VP and LP, there is a 70% reduction of the total number of cells, whereas the reduction is less in DP and CG. It thus seems that the reduction of prostatic epithelial tissue mass upon castration is due to a reduction of the number of cells as well as a reduction of the volume of individual cells. VP and LP appear to be more androgen-dependent than DP and CG.     

Prospective comparison of hip fracture treatment: 856 cases followed for 4 months in The Netherlands and Sweden

In a prospective multicenter study 1115 hip fracture patients were registered in Rotterdam (The Netherlands), Sundsvall and Lund (Sweden). The patients had similar background parameters with a mean age of 78 years, about half of them living alone and just above 80 percent coming from independent living

For cervical fracture, hemiarthroplasty was the predominating treatment in Rotterdam (n 169), whereas osteosynthesis was used in Sundsvall (screws n 135) and Lund (hook-pins n 148). The mean (median) hospitalization time was 32 (20) days in Rotterdam, 16 (12) days in Sundsvall, and 17 (10) days in Lund. Discharge to independent living varied from 53 percent in Lund to 72 percent in Sundsvall. Functional outcome (walking ability and ADL capacity) was at 4 months similar in all groups, but at 2 weeks was lower in Rotterdam. Mortality at 2 weeks / 1 month 14 months was in Rotterdam 4/9/20, in Sundsvall 2/4/13, and in Lund 01311 0 percent.

Trochanteric fractures were treated by screwplate in Rotterdam (n 146) and Lund (n 78), and by Ender nails in Sundsvall (n 11 7). The mean (median) hospitalization time was in Rotterdam 39 (29) days, in Sundsvall 24 (1 5) days and in Lund 19 (11) days. Discharge to independent living varied from 41 percent in Lund to 57 percent in Sundsvall. Functional outcome was similar between the groups. Mortality at 2 weeks / 1 month 14 months was in Rotterdam 2/6/14, in Sundsvall 611 2/19 and in Lund 12/12/18 percent.

Thus, our study has shown that it is possible to perform a prospective multicenter study involving different European countries. The functional out- come after 4 months was very consistent between the centers studied, irrespective of choices made concerning operation method and rehabilitation routines. However, a difference in mortality within these first postoperative months was found, which seems attributable to the operation procedure.