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Biomedical subjects

M Larsen

Publications and source records attributed to M Larsen.

At least 55 records · Page 3Linked to original sources

Diabetic macular edema: passive and active transport of fluorescein through the blood-retina barrier.

PURPOSE: To investigate the passive bidirectional and active outward transport of fluorescein through the blood-retina barrier (BRB) in diabetic patients with clinically significant macular edema and in healthy controls. METHODS: The passive and active transport of fluorescein through the BRB was quantitated by vitreous fluorometry. A previously developed method was used to model passive transport. A new simulation model was developed and evaluated for estimation of active transport. The study included 10 eyes of 5 healthy controls and 31 eyes of 20 diabetic patients with clinically significant diabetic macular edema (CSME) in at least one eye, totalling 25 eyes with CSME. RESULTS: Passive permeability of fluorescein was increased by a factor of 12 in eyes with edema compared to healthy controls (edema, 23.7 nm/sec; healthy subjects, 1.9 nm/sec, P < 0.01), whereas the active transport was doubled (edema, 84.1 nm/sec; healthy subjects, 43.5 nm/sec, P < 0.01). Unlike active transport, passive permeability was related to the degree of retinopathy, in that eyes with severe non-proliferative diabetic retinopathy had a passive permeability that was significantly increased compared to moderate retinopathy (32.1 nm/sec and 14.6 nm/sec, respectively, P: < 0.05). The passive movement quantitated with vitreous fluorometry was larger for diffuse and mixed leakage compared to focal (P = 0.07). CONCLUSIONS: Insofar as the movement of fluorescein can be taken as a probe for the movement of electrolytes and water, the pathogenesis of diabetic macular edema seems to involve a disruption of the BRB, presumably its inner component. The active resorptive functions of the blood-retina barrier appear to be compensatorily increased to counteract edema formation, although the increase is too small to prevent edema in the face of severe leakage through the blood-retina barrier.

Adult↗

Effect of different times of administration of the nematophagous fungus Duddingtonia flagrans on the transmission of ovine parasitic nematodes on pasture--a plot study.

Investigations were made into the timing of administration of Duddingtonia flagrans as a biological control agent against ovine parasitic nematodes including stongylid and Nematodirus spp. Faeces from 3-4 months old male lambs were deposited onto pasture plots that had never been grazed by sheep. The trial was conducted over two consecutive years (1998 and 1999). For both years, the following three plot types were involved: Sim plots had faeces containing nematode eggs and Duddingtonia flagrans spores deposited simultaneously; Post plots had faeces containing nematode eggs followed 2 weeks later by faeces containing D. flagrans spores alone; Control plots had faeces containing only nematode eggs; Prior plots (included in 1999) had faeces containing D. flagrans spores alone followed 2 weeks later by faeces containing nematode eggs. In each year, two deposition periods were involved: July and August in 1998 and June and July in 1999. During the first year pasture samples were collected at 2, 4, 6, 8 and 12 weeks after initial deposition. In 1999, additional samples were collected at 10, 16 and 20 weeks. Larvae were extracted from the pasture samples and counts performed to estimate the number and species of infective third-stage (L(3), larvae) present. The number of third-stage strongylid larvae on pasture was significantly lower on Sim plots compared to the remaining plot types for both years at all deposition times (P<0.001). This was also the case for the number of Nematodirus infective larvae in August deposition plots in 1998 (P<0. 02). There was no significant difference between treatments in both deposition times in 1999 and July deposition plots in 1998 for the Nematodirus data. These results suggest that D. flagrans, if deposited at the same time as parasite eggs prevents transmission of third-stage larvae from the faecal deposit onto pasture, including occasionally Nematodirus species, but does not have an effect on third-stage parasitic nematode larvae in the surrounding soil.

Animals↗

Efficacy of the nematophagous fungus Duddingtonia flagrans in reducing equine cyathostome larvae on pasture in south Louisiana.

The effectiveness of Duddingtonia flagrans in reducing the free living third stage larvae (L(3)) of equine cyathostomes on pasture when fed to horses has been demonstrated in cold temperate climates. The objective of this experiment was to assess the efficacy of D. flagrans against equine cyathostomes in the subtropical environment of southern Louisiana. Fecal pats were prepared by mixing feces obtained from a parasite-free horse fed D. flagrans at a dose of approximately 2 x 10(6) spores kg(-1), with feces containing cyathostome eggs from a parasitized horse. Control pats contained feces from a parasite-free horse mixed with feces containing cyathostome eggs. The fecal pats were placed on pasture in six replicates at 4-week intervals from March 1997 until January 1998. Comparison of recoveries of L(3) from non-treated control pats in the field with non-treated coprocultures maintained in the laboratory indicated that L(3) survival on pasture was reduced during the months of May, June, July, August and September. The efficacy of the fungus was determined by L(3) recovery from grass surrounding the fecal pats of treated and control groups. D. flagrans significantly reduced L(3) during the months of April, May, and October 1997 to January 1998 (range 66-99% reduction, p=0.0001), and for the year as a whole (p=0.0001).

Animals↗

Kluyveromyces lactis killer system: identification of a new gene encoded by pGKL2.

The cytoplasmic linear plasmid pGKL2 of the yeast Kluyveromyces lactis was reported to harbour ten open reading frames (ORF1-ORF10). By re-examining the sequence, we identified an additional ORF encoding a polypeptide of 70 amino acids; and a homologous ORF with 70% similarity was also identified on plasmid pSKL of Saccharomyces kluyveri. As for pGKL2, the newly detected ORF11 is located at the same position with the same direction. ORF11 transcripts of pGKL2 were verified by reverse transcriptase-polymerase chain reaction; and the corresponding promoter activity was proven by expression of a reporter gene driven by the ORF11 upstream conserved sequence, indicating that ORF11 constitutes a functional gene.

Amino Acid Sequence↗

The WFDC1 gene encoding ps20 localizes to 16q24, a region of LOH in multiple cancers.

We previously identified ps20 protein as a secreted growth inhibitor and purified the protein from fetal rat prostate urogenital sinus mesenchymal cell conditioned medium. The rat cDNA was subsequently cloned, and ps20 was found to contain a WAP-type four-disulfide core motif, indicating it may function as a protease inhibitor. We now report cloning and characterization of the mouse ps20 gene (designated Wfdc1), the human homolog cDNA, and the human gene (designated WFDC1). Both the mouse and human WFDC1 genes consist of seven exons and encode respective ps20 proteins sharing 79.1% identity and nearly identical WAP motifs in exon 2. The WFDC1 gene was mapped by FISH analysis to human Chromosome (Chr) 16q24, an area of frequent loss of heterozygosity (LOH) previously identified in multiple cancers including prostate, breast, hepatocellular, and Wilms' tumor. Identification and characterization of the WFDC1 gene may aid in better understanding the potential role of this gene and ps20 in prostate biology and carcinogenesis.

Amino Acid Sequence↗

Monofocal outer retinitis associated with hand, foot, and mouth disease caused by coxsackievirus.

PURPOSE: To report symptoms and signs from an otherwise healthy man with photopsia and monofocal outer retinitis occurring 3 days after the onset of hand, foot, and mouth disease. METHOD: Case report. RESULTS: Ophthalmic examination revealed a single circumscribed area of pigment epithelial mottling in the left eye and a corresponding nonarcuate absolute scotoma. Fluorescein angiography demonstrated attenuation of the retinal pigment epithelium. Serologic tests at 1 month after the onset disclosed serum antibody to enterovirus. CONCLUSION: The close association between the cutaneous eruption and the onset of visual symptoms suggests that the retinal lesion was caused by coxsackievirus infection.

Adult↗

Biological control of trichostrongylid infections in calves on pasture in Lithuania using Duddingtonia flagrans, a nematode-trapping fungus.

The effect on the pasture contamination level with infective trichostrongylid larvae by feeding the nematode-trapping fungus, Duddingtonia flagrans at two dose levels to first time grazing calves was examined in Lithuania. Thirty heifer-calves, aged 3-6 months, were divided into three comparable groups, A, B and C. Each group was turned out on a 1.07 ha paddock (a, b and c). The paddocks were naturally contaminated with infective trichostrongylid larvae from infected cattle grazing the previous year. Fungal material was fed to the animals daily during a two month period starting 3 weeks after turnout. Groups A and B were given 10(6) and 2.5x10(5) chlamydospores per kg of live weight per day, respectively, while group C served as a non-dosed control group. Every two weeks the heifers were weighed and clinically inspected. On the same dates, faeces, blood and grass samples were collected. From mid-July onwards, the number of infective larvae in grass samples increased markedly (P<0.05)on paddock c, whereas low numbers of infective larvae were observed on paddocks a and b grazed by the fungus treated groups. However, the results indicate that administering fungal spores at a dose of 2.5x10(6)chlamydospores per kg live weight per day did not significantly prevent parasitism in calves, presumably due to insufficient suppression of developing infective larvae in the faeces. In contrast, a dose of 10(6) chlamydospores per kg lowered the parasite larval population on the pasture, reduced pepsinogen levels (P<0.05), and prevented calves from developing parasitosis.

Animals↗

Prospects for controlling animal parasitic nematodes by predacious micro fungi.

Resistance against anthelmintics is widespread, particularly in parasitic nematode populations of small ruminants. Several new techniques or supplements have been developed or are under investigation. Biological control (BC) is one of these new methods. The net-trapping predacious fungus Duddingtonia flagrans produces thick walled resting spores, chlamydospores, which are able to survive passage through the gastrointestinal tract of cattle, horses, sheep and pigs. Under Danish climatic conditions it has been shown that the number of parasite larvae on pasture and the worm burden of the grazing animals is significantly reduced when animals are fed spores during the initial 2-3 months of the grazing season. Work with D. flagrans in France, Australia, USA, and Mexico has confirmed the strong BC potential of this fungus. Today much work is going into development of suitable delivery systems for grazing livestock worldwide. Ultimately, BC should be implemented in integrated parasite control strategies, both in conventional and organic livestock production.

Animals↗

High rate of GB virus type C/HGV transmission from mother to infant: possible implications for the prevalence of infection in blood donors.

BACKGROUND: Because GB virus type C(GBV-C)/HGV (GBV-C/HGV) is blood-borne and sexually transmitted, persons at risk of infection with such viruses have a high prevalence of GBV-C/HGV markers. However, adults with no apparent risk factors, such as blood donors, frequently are positive for GBV-C/HGV markers. Mother-to-infant transmission could explain this high prevalence, but it has been studied only through small series of GBV-C/HGV-infected mothers co-infected with HCV or HIV. STUDY DESIGN AND METHODS: To determine the rate of mother-to-infant transmission of GBV-C/HGV RNA in women who are HCV- or HIV-negative, a prospective study was performed in a cohort of 288 mothers screened for viral RNA and in the infants born to GBV-C/HGV-infected mothers. RESULTS: Thirteen mothers (4.5%) were found positive for GBV-C/HGV RNA. Of the infants in whom at least one blood sample was collected between the third and the ninth months of life, 89 percent were positive for viral RNA. The majority of these newborns were negative for GBV-C/HGV RNA at birth and positive after the third month. The viral RNA titers of infants born to GBV-C/HGV-infected mothers appeared as elevated as those of their mothers. All the GBV-C/HGV-infected infants remained positive for viral RNA during the entire study period. No clinical events possibly linked to a primary GBV-C/HGV infection were reported in infants. Serum ALT level and blood count remained within normal values throughout the follow-up of all GBV-C/HGV-infected infants. CONCLUSION: The frequency of mother-to-infant GBV-C/HGV transmission is elevated and could explain the high prevalence of GBV-C/HGV markers (viral RNA and E2 antibody) in adults at low risk for blood-borne or sexually transmitted viruses, such as blood donors.

Adult↗

Absence of obvious short-term impact of the nematode-trapping fungus Duddingtonia flagrans on survival and growth of the earthworm Aporrectodea longa.

The nematode-trapping fungus Duddingtonia flagrans may be used in biological control of parasitic nematode larvae in faeces of domestic host animals after feeding the hosts with fungal chlamydospores. In this experiment a possible undesirable fungal impact on earthworms, of the species Aporrectodea longa, was investigated. As earthworms eat animal faeces, D. flagrans may come into contact with earthworms both in their alimentary tract and on their body surface. However during the experimental period of 20 days, when earthworms were living in soil and eating cattle faeces that were heavily infested with viable chlamydospores of D. flagrans there were no indications of internal or external mycosis among the earthworms.

Animals↗

The URA5 gene encoding orotate-phosphoribosyl transferase of the yeast Kluyveromyces lactis: cloning, sequencing and use as a selectable marker.

A pair of degenerate primers was used for amplification and cloning of an internal fragment of the K. lactis URA5 gene. Primers were designed on the basis of highly conserved motifs within protein sequences predicted for URA5 genes from several microorganisms. Using the amplified fragment as a probe, we finally cloned and sequenced a 1.9 kb chromosomal fragment containing the orotate-phosphoribosyltransferase-encoding URA5 gene and an incomplete open reading frame strikingly similar to SEC65 of Saccharomyces cerevisiae and other yeasts, in which the gene encodes a subunit of the signal recognition particle. Uracil-requiring mutants of K. lactis CBS 683 were selected on media containing 5-fluoro-orotic acid and used as recipients in transformation experiments using K. lactis URA5 as the selectable marker, thereby proving functionality of the cloned gene.

Amino Acid Sequence↗

The efficacy of two isolates of the nematode-trapping fungus Duddingtonia flagrans against Dictyocaulus viviparus larvae in faeces.

A series of experiments was carried out to examine the effects of two different isolates of the nematode-trapping fungus Duddingtonia flagrans to reduce the number of free-living larvae of the bovine lungworm, Dictyocaulus viviparus. A laboratory dose-titration assay showed that isolates CI3 and Troll A of D. flagrans significantly reduced (P < 0.05 to P < 0.001) the number of infective D. viviparus larvae in cultures at dose-levels of 6250 and 12,500 chlamydospores/g of faeces. The larval reduction capacity was significantly higher for Troll A compared to CI3 when lungworm larvae were mixed in faecal cultures with eggs of Cooperia oncophora or Ostertagia ostertagi and treated with 6250 chlamydospores/g of faeces. Both fungal isolates showed a stronger effect on gastrointestinal larvae than on lungworm larvae. Two plot trials conducted in 1996 and 1997 involved deposition of artificial faecal pats containing free-living stages of D. viviparus and C. oncophora on grass plots. Herbage around the pats was collected at regular intervals and infective larvae recovered, counted and identified. These experiments showed that both D. flagrans isolates reduced the number of gastrointestinal as well as lungworm larvae in faecal pats. During both plot trials, the transmission of C. oncophora larvae, but not D. viviparus, from faecal pats to the surrounding herbage was clearly affected by climatic conditions. After collection of faecal pats from the grass plots one month after deposition, the wet and dry weight of pats as well as organic matter content were determined. No differences were found between the fungus-treated and non-treated control pats. This indicated that the rate of degradation of faeces was not affected by the addition of the fungus.

Animals↗

Integrated and biological control of parasites in organic and conventional production systems.

Organic and other non-intensive animal production systems are of growing importance in several countries worldwide. In contrast to conventional farms, parasite control on organic farms is affected by several of the prescribed changes in management e.g. access to the outdoors in the summer and in most countries, a ban on preventive medication, including use of anti-parasiticides. Organic animal production relies heavily on grazing, and pasture or soil related parasites are thus of major importance. Several studies in northern temperate climate have indicated that outdoor production of pigs, primarily sows, and laying hens results in heavier and more prevalent helminth infections compared to conventional intensive production under indoor conditions. In organic dairy cattle, parasitic gastroenteritis in heifers may be more prevalent. In a short to medium term perspective, integrated control may combine grazing management with biological control using nematophagous micro-fungi, selected crops like tanniferous plants and on conventional farms, limited use of anti-parasiticides. At present, the non-chemotherapeutic control of pasture related infections is based mainly on grazing management strategies. Preventive strategies, where young, previously unexposed stock, are turned out on parasite-free pastures, can be used for grazing first season dairy heifers and in all-in-all-out poultry production. Evasive strategies aim at avoiding disease producing infections of a contaminated area by moving to a clean area and may be relevant for ruminants and pigs. In cattle, effective control of nematodes can be achieved by repeated moves of the herd or alternate grazing with other species. High stocking rates seem to be an important risk factor. In pig production, the effect of paddock rotation on parasite infections is largely unknown and studies are warranted. Control of nematodes by larvae-trapping fungi, or perhaps in the future by egg-destroying fungi, looks promising for ruminants and certain monogastric animals but delivery systems and practical dosing regimes integrated with grazing management have to be developed. In conclusion, good prospects are expected for acceptable parasite control without a heavy reliance on anti-parasiticides through integration of the above mentioned procedures but future studies are needed to confirm their efficacy under practical farming conditions.

Animal Husbandry↗

Growth rate and trapping efficacy of nematode-trapping fungi under constant and fluctuating temperatures.

The effect of temperature on radial growth and predatory activity of different isolates of nematode-trapping fungi was assessed. Four isolates of Duddingtonia flagrans and one isolate of Arthrobotrys oligospora were inoculated on petri dishes containing either cornmeal agar (CMA) or faecal agar and then incubated for 14 days under three different constant and fluctuating temperature regimes. The radial growth was similar on the two substrates at each temperature regime. All fungal isolates showed a higher growth rate at a constant 20 degrees C. At 10 degrees and 15 degrees C, all D. flagrans isolates showed very similar patterns of radial growth at both constant and fluctuating temperatures. At 20 degrees C, they grew significantly faster at constant than at fluctuating temperatures. A. oligospora grew significantly faster than all D. flagrans isolates except when incubated at a fluctuating 20 degrees C. Spores of each fungal isolate were added to faecal cultures containing eggs of Cooperia oncophora at a concentration of 6250 spores/g faeces. The cultures were incubated for 14 days at the same temperature regimes described above. Control faeces (without fungal material) were also cultured. More larvae were recovered from the fungus-treated cultures incubated at a constant 10 degrees or 15 degrees C than from those incubated at the respective fluctuating temperatures, except for one D. flagrans isolate. Incubation at 20 degrees C showed the opposite effect. The general reduction observed in the number of nematode larvae due to fungal trapping was 18-25% and 48-80% for a constant and fluctuating 10 degrees C, 70-96% and 93-95% for a constant and fluctuating 15 degrees C, and 63-98% and 0-25% for a constant and fluctuating 20 degrees C, respectively.

Animals↗

Biological control of helminths.

As a potential component in future integrated parasite-control strategies, biological control by means of predacious fungi seems to be moving from a promising possibility toward becoming a reality, and the netforming nematode-destroying fungus Duddingtonia flagrans appears to be the candidate of choice. Not only has this fungus been found in, and isolated from, fresh sheep, cattle and horse faeces, but it also appears to be the only fungus that is able to consistently and significantly reduce the number of infective trichostrongyle larvae in faeces from animals fed fungal spores. Results from the last few years have shown that D. flagrans is able to trap and destroy free-living stages of the most important and common trichostrongylid larvae with very similar external life-cycles, as well as larvae of parasites with a slightly different transmission biology (Nematodirus spp., Dictyocaulus viviparus). The introduction of microfungi for biological control could be as part of a feed supplement or incorporated in feed-blocks presented to animals which are raised under relatively intensive conditions and constant surveillance. Apart from the special niche for organic farmers, such a product would be suited for horses, small ruminants (as either milking herds or housed daily for other reasons), cattle in special situations and free-roaming pigs. The most important constraint, still, for a major breakthrough in biological control in the latter is the lack of good antagonists against the long-lived and rather resistant infective stages of parasites, being transmitted as larvae inside the egg. Since the first Conference on Novel Approaches to the Control of Helminth Parasites of Livestock in Armidale, Australia, 1995, there has been a steady evolution within the area of biological control of parasitic nematodes. Today this principle is being exploited and tested out in almost all parts of the world, under various climatic conditions and production systems. Where, in the past, a large part of the work focused on cattle and to a lesser degree horse and sheep parasites, the focus of the research in many of the newly involved countries is on small ruminants, because of their importance to primarily small-scale farmers in local communities. Today research and trials are either on-going or being planned in many developing countries, as well as in countries in transition. The involvement of multinational agencies in addition to national and industrial interests is very welcome and should increase the chances and keep up the momentum for development and implementation of biological control in future animal production around the world.

Animals↗

Pathophysiology and hemodynamics of branch retinal vein occlusion.

PURPOSE: To describe the pathophysiologic and the hemodynamic changes associated with branch retinal vein occlusion (BRVO) on the basis of selected angiographic observations from a cohort of patients with BRVO and related anomalies of retinal blood flow. DESIGN: Retrospective, observational case series. PARTICIPANTS: A total of 250 patients with incipient or manifest BRVO and 5 patients with related anomalies. METHODS: Color and red-free gray-scale fundus photography and intravenous fluorescein angiography. MAIN OUTCOME MEASURES: Morphologic signs of disturbed retinal blood flow. RESULTS: All occlusions occurred at arteriovenous crossing sites where the artery is positioned anterior to the vein. Presumptive precursor abnormalities of blood flow at arteriovenous crossings include turbulence and upstream venous dilation. After the onset of BRVO, the clinical course is determined by the location of the BRVO in relation to the fovea, the extent of the involved venous drainage area, and the collateral drainage capacity from the area with compromised venous drainage to the adjacent areas of intact venous drainage. Collateral maturation occurs over a period of 6 to 24 months after the onset of BRVO, when a transient retinal edema may be seen that has a preponderance for foveal involvement because the perifoveal area has the highest density of preformed collaterals between adjacent venous drainage areas. CONCLUSIONS: The treatment of BRVO may possibly benefit from a refined angiographic analysis of the process of collateral formation and new treatment methods aimed at accelerating the process of collateral maturation.

Acute Disease↗

[Retrospective study of exchange transfusion for newborn infants with reconstituted blood. Review of 60 exchanges].

PURPOSE: The necessity of saving blood products has established the practice of exchange transfusion (ET) with reconstituted blood in newborns. The aim of this retrospective study was to evaluate the indications and the practice of this technique at the Perinatal Hemobiology Centre (Paris, France). METHODS: The records of intervention allowed us to review the etiologic categories for neonates having undergone exchange transfusion with reconstituted blood, the dosages used (bilirubin, hemoglobin), and the other main parameters of ET. RESULTS: Sixty ETs were performed in 48 newborns between the 1st July 1996 and the 1st July 1998. Twenty-seven with Rh hemolytic disease had 39 ETs (19 for hyperbilirubinemia, 12 for anemia, and eight for both), whereas ten out of 12 repeated ETs were indicated for hyperbilirubinemia (six of these cases were in newborns weighing > or = 2500 g and after a volume exchange < or = 1 blood mass [range 0.72-1.0] at the last ET). Twenty-one cases showed other diseases: six of them had anemia, nine had hyperbilirubinemia, and seven showed disseminated coagulopathy. The tolerance of ET was poor in 24% infants in this group. CONCLUSIONS: The volume of 1.3 blood mass for ET is sufficient for the majority of cases with hyperbilirubinemia, allowing transfusional savings in comparison with the previous recommendation of two blood volumes. Exact labeling of the content of units of packed red cells and plasma is essential to fulfill the volume and hematocrit requirements in every case.

Anemia↗

Effect of Duddingtonia flagrans against Ostertagia ostertagi in cattle grazing at different stocking rates.

The efficacy of an isolate of the nematophagous fungus Duddingtonia flagrans against gastrointestinal nematodes of cattle was examined at 2 dose levels on 2 permanent pastures, with high and low stocking rates, respectively. Thirty calves, experimentally infected with Ostertagia ostertagi, were divided into 3 comparable groups and allocated to 3 similar paddocks in each of the 2 trials. Two of the 3 groups received fungal material once per day during the initial 2 months, either at high dose (10(6) fungal spores/kg body weight) or low dose (5 x 10(5) or 2.5 x 10(5) fungal spores/kg body weight). The third group remained as an untreated control group. Faecal, blood, and herbage samples were collected and animals were weighed every month from May to September. The pasture grazed at a high stocking rate had a large number of overwintering infective larvae, while the pasture grazed at a low stocking rate had a low overwintering herbage larval infectivity. The results showed that, at a high stocking rate, the recovery of infective larvae on pasture was diminished and calves were prevented from clinical ostertagiosis by using the D. flagrans Troll A-isolate. At low stocking rate, the parasite burden seemed not to be very heavy, and a conclusive effect of the fungi at the dose-level used could not be detected.

Animal Husbandry↗