Rehydration methods to recover cysticercoids of the rat tapeworm Hymenolepis diminuta from dry flour beetle carcasses.

Terrestrial arthropods host a variety of helminth parasites, yet quantifying the intensity of infection in these hosts post-mortem is challenging because carcasses may desiccate quickly. We recovered cysticercoids of Hymenolepis diminuta from desiccated flour beetle (Tribolium confusum) carcasses by modifying a published insect rehydration procedure. Without rehydration, carcasses dissected more than 1 day post-mortem had noticeable degradation of cysticercoids. Mild rehydration (soaking in water only for 2 days, or 0.5-10% KOH for 1 h followed by 1 day in water, or 0.5% KOH for 1 day) left carcasses tough and time-consuming to dissect, but all parasites could be recovered and were similar in body size to fresh cysticercoids. Moderate rehydration (5-10% KOH for 1 day) allowed all parasites to be recovered and facilitated dissection by partially dissolving internal organs of the beetle while causing little degradation of the cysticercoids. Harsh rehydration (5-10% KOH for 1 day followed by 5 days in water) not only dissolved internal beetle tissues but also severely damaged cysticercoids, such that parasite counts were unreliable. The degree of initial carcass desiccation had little effect on results following rehydration. However, regardless of treatment used, intact cercomers were rarely retained on rehydrated cysticercoids. Rehydration was less successful on early developmental stages of the parasite, which were recovered reliably only as they neared the cysticercoid stage. This method has utility for studies of parasite-induced mortality by permitting accurate and reliable parasite counts from dead, desiccated hosts.

Islet-enriched gene expression and glucose-induced insulin secretion in human and mouse islets.

AIMS/HYPOTHESIS: Our understanding of the transcription factors that control the development and function of rodent islet beta cells is advancing rapidly, yet less is known of the role they play in similar processes in human islets. METHODS: To characterise the abundance and regulation of key proteins involved in glucose-regulated insulin secretion in human islets, we examined the expression of MAFA, MAFB, GLUT2 (also known as SLC2A2), ßGK (also known as GCK) and PDX1 in isolated, highly purified human islets with an intact insulin secretory pattern. We also assessed these features in islets from two different mouse strains (C57BL/6J and FVB). RESULTS: Compared with mouse islets, human islets secreted more insulin at baseline glucose (5.6 mmol/l), but less upon stimulation with high glucose (16.7 mmol/l) or high glucose plus 3-isobutyl-1-methyl-xanthine. Human islets had relatively more MAFB than PDX1 mRNA, while mouse islets had relatively more Pdx1 than Mafb mRNA. However, v-maf musculoaponeurotic fibrosarcoma oncogene homologue (MAF) B protein was found in human islet alpha and beta cells. This is unusual as this regulator is only produced in islet alpha cells in adult mice. The expression of insulin, MAFA, ßGK and PDX1 was not glucose-regulated in human islets with an intact insulin secretory pattern. CONCLUSIONS/INTERPRETATION: Our results suggest that human islets have a distinctive distribution and function of key regulators of the glucose-stimulated insulin secretion pathway, emphasising the urgent need to understand the processes that regulate human islet beta cell function.

Manipulation of host food availability and use of multiple exposures to assess the crowding effect on Hymenolepis diminuta in Tribolium confusum.

We studied the 'crowding effect' in Tribolium confusum infected with Hymenolepis diminuta. Manipulations included age and number of parasites, and diet, sex, age and number of exposures of hosts. Volume per parasite was unaffected until an intensity of at least 5-10 parasites per host, then declined approximately inversely as intensities increased. Parasite size was affected by host sex but not age or reproductive status. Host diet affected parasite size and the impact of crowding. Daily gain in parasite volume peaked partway through the developmental period and preceded the first evidence of a crowding effect. Parasites that established during a second exposure had a transient developmental delay but eventually grew as large or larger than parasites from a single exposure with the same total intensity. Parasites responded to crowding by differential allocation of resources. Cercomer volume decreased even with slight crowding, the capsule surrounding the scolex was not reduced until crowding became more severe, and scolex width was reduced only in the most extreme conditions. The data support the hypothesis that the crowding effect in this system is driven primarily by nutrient, rather than space limitations.

A short review of experimental peritoneal sclerosis: from mice to men.

Peritoneal sclerosis has been induced in rodents in vivo by exposing the membrane to a variety of experimental interventions: asbestos, 0.1% chlorexidine, iron dextran, glucose degradation products, AGE deposits derived from uremia per se, sodium hypochlorite, lypopolysaccharide, low pH, pure water, silica or zymosan. With a few exceptions (pure water, chlorhexidine and low pH), the other substances mentioned operate setting out different degrees of oxidative stress. This short review describes several experimental interventions in rodents, aimed at acute exfoliation or long-term, sustained injury of the mesothelial monolayer performed by means of intraperitoneal injections of different oxidant agents. Acute exfoliation induced by deoxycholate resulted in a depopulated monolayer coincident with immediate alteration of the peritoneal permeability, evidenced by increased urea D/P ratio, higher glucose absorption rate, elevated albumin losses in the effluent and significant reduction of the ultrafiltration rate. In the long term (30 days), these manifestations of membrane failure persisted and coincided with substantial peritoneal sclerosis. Peritoneal sclerosis was also induced by IP injections of 0.125% trypsin and 6.6 mM/L solution of formaldehyde. Using the doughnut rat model of mesothelial regeneration, exposure to 4.25% glucose or 7.5% icodextrin solutions severely hampered repopulation of the monolayer, which was replaced by a thick sheet of fibrous tissue. It is concluded that peritoneal sclerosis derives mostly from sustained oxidative injury to the peritoneal membrane. Loss of the mesothelial monolayer is the first step in the chain of events leading to this complication.

Temperature effects on survival and excystment of cercariae of Halipegus occidualis (Trematoda).

Survival of the cystophorous cercariae of Halipegus occidualis (Trematoda: Hemiuridae) from naturally infected Helisoma anceps (Gastropoda: Planorbidae) was determined following constant temperature storage at 4, 16, 22, 26 and 30 degrees C. At 4 degrees C, 90% of cercariae survived for 14 weeks and 50% survived for 30 weeks. Survival varied inversely with temperature, and at 30 degrees, 90% survived for 1.5 weeks and 50% survived for 2.5 weeks. At the time of release from snails, cercariae excysted within 0.5 s of stimulation. Excystment times ultimately increased to 1-3 s at all temperatures as length of storage increased, but no connection was detected between excystment time and the activity of the cercaria. Excystment times began increasing while survival was still above 90%. Temperature specific mortality rates of cercariae were estimated, and used to simulate the survival of cercariae released naturally at various times of the year. The results of these simulations suggest that cercariae of H. occidualis are present in the environment at all times of the year, but that cercariae present in late winter (January to mid-March) may have an impaired excystment mechanism that reduces their infectivity.

Increased mesenteric, diaphragmatic, and pancreatic interstitial albumin content in rats with acute abdominal sepsis.

This study was designed to evaluate the decreased permselectivity of the capillary wall and the resultant higher permeability to macromolecular anionic albumin in septic rats, by quantitative estimation of Evans blue-albumin complexes in interstitial tissue. Septic peritonitis was induced by intraperitoneal injection of Escherichia coli-O6 KS H16. Twenty-four hours after induction of septic peritonitis, intact (healthy, noninoculated animals) and septic rats were perfused with 5 mL/Kg of a solution of Evans blue in normal saline (20 mg/mL in .9% NaCl). In septic rats, the interstitial concentration of Evans blue in mesentery, pancreas, and diaphragmatic muscle was significantly higher than that observed in intact animals. The present observations were made in the same experimental model of abdominal sepsis that showed a substantial reduction in the endothelial negative charge of the mesenteric, pancreatic, and diaphragmatic capillary beds. The evidence obtained from this experiment confirms that the loss of the permselective properties of capillary wall for macromolecular anionic albumin derives from a drastic reduction of its normally present and regularly distributed fixed electronegative charges.

Treatment of surgical and non-surgical septic multiorgan failure with bicarbonate hemodialysis and sequential hemofiltration.

OBJECTIVE: Hospital mortality of patients with septic multiorgan failure (MOF) is still around 95%. The present study investigates whether this high mortality could be significantly reduced by the addition of sequential hemofiltration (SH) with bicarbonate hemodialysis (HD) to the currently used life supportive measures. DESIGN: 35 (18 surgical and 17 nonsurgical) patients, with 3 or more organ failures, had daily sessions of zero balance SH, for periods ranging from 2-22 days. MEASUREMENTS AND RESULTS: SH induced significant improvement of PaO2/100 FIO2, Apache II score, MAP, as well as blood chemistry in survivors. Dying patients had less marked improvement of blood oxygenation, non-significant changes in other variables, in addition to low MAP before and after SH, as well as marked hemodynamic unstability during the procedure. The observed hospital mortality was 38% for the surgical group, and 35.3% for the medical patients (n.s.). CONCLUSIONS: Mortality observed in this retrospective, uncontrolled study was significantly lower than that currently observed with conventional supportive therapy, with or without the addition of other forms of blood purification, e.g. CAVH and CAVHD. This improvement in results appears to be related to the property of SH to completely clear 90% of the blood from mediators of inflammation in only one passage through the hemofilter, and to better tolerance of HD done using bicarbonate buffer. A definite evaluation of this technique will be eventually reached by a programmed, appropriate sample size study, which is out of reach for one individual ICU.

Climbing simulated vegetation to heights of ungulate hosts by larvae of Dermacentor albipictus (Acari: Ixodidae).

Larvae of winter ticks, Dermacentor albipictus (Packard), ascend vegetation in autumn and form clumps that attach to passing ungulate hosts. We tested the hypothesis that vegetation height determines the height of clumps. During the vegetation-to-ungulate transmission period (early September to mid-November), larvae were released at the base of simulated vegetation (nylon rods 245 cm tall) in outdoor and laboratory trials and in the absence of host cues. Rod height exceeded the height of the tallest ungulate host, which is the moose, Alces alces (L.). Most larvae stopped climbing and formed clumps 50-190 cm above ground, which coincided with torso heights of moose; elk, Cervus elaphus L.; and deer, Odocoileus spp. Rafinesque. More clumps formed in outdoor trials than in laboratory trials and clump heights tended to increase over the course of the experiment, but clump number, size, and height did not correlate with weather conditions. Winter tick larvae appear to determine their height above ground in the absence of external cues, but this mechanism may be modified by external conditions.

Application of electron spin resonance for evaluation of the level of free radicals in the myometrium in full-term pregnancy with normal labour and uterine inertia.

In order to identify and quantify free radicals in the tissues of patients with normal physiological and pathological states of births, we developed a method to evaluate the amount of free radicals in myometrium of subplacental area and from body of uterus, using electron spin resonance spectroscopy. Analysis of the concentration of free radicals in the myometrium in full-term pregnancy with normal labour and during uterine inertia was studied. The activities of Ca2+-ATPase, cytochrome c oxidase and succinate dehydrogenase in samples of these tissues were tested too. Low free radical concentrations in these tissues were associated with disturbances in contractile activity of myometrium along with reduction of Ca2+-ATPase, cytochrome c oxidase and succinate dehydrogenase activity. There proved to be an association between the level of free radicals in the tissues and alteration in the physiological processes.

The Ural train-gas pipeline catastrophe: the report of the IDF medical corps assistance.

Following the destruction of two trains in the Urals 2000 km east of Moscow, as a consequence of the conflagration caused by an explosion from a leaking natural gas pipeline, 3000 people were injured;* most of them (2200) died* immediately and the others (about 800) were badly burned. At the request of the Soviet Union Government a medical military delegation was sent to give assistance to the injured people. This report describes the treatment given by the delegation to 40 patients with burns of between 40 and 90 per cent TBSA during a period of 10 days. An insight into a Soviet Union Trauma Center is given and the good treatment given by the Soviet colleagues is emphasized.

Helminth position within the intestine of naturally infected pike (Esox lucius) relative to host stomach contents.

The positions of 3 cestode species, Triaenophorus crassus Forel, Triaenophorus nodulosus (Pallas), and Proteocephalus pinguis LaRue, and a nematode, Raphidascaris acus (Bloch), within the intestine of naturally infected pike (Esox lucius L.) were evaluated with respect to the presence or absence and state of digestion of host stomach contents. The positions of scolices and strobilae of the cestodes did not vary with respect to host stomach contents. By contrast, R. acus was anterior (near the stomach) when the stomach contained partially digested items, posterior when the stomach was empty, and in an intermediate position when the stomach contained only intact items. These results are interpreted as migration of R. acus, but not T. crassus, T. nodulosus, or P. pinguis, in response to feeding activity of the host. Adult and larval R. acus migrated, but the extent of migration was reduced in hosts harboring T. crassus, more so for larval than adult R. acus.

Moisture and temperature effects on survival and infectivity of first-stage larvae of Parelaphostrongylus odocoilei and P. tenuis (Nematoda: Metastrongyloidea).

The survival of first-stage larvae (L1) of Parelaphostrongylus odocoilei and P. tenuis (Nematoda: Metastrongyloidea) and their infectivity to the snail Triodopsis multilineata were determined experimentally in a variety of temperature and moisture conditions. Survival of larvae of P. odocoilei increased with decreasing temperature. Survival of larvae in water was similar to survival in air at 17 and 45% RH; survival of larvae in air at 75, 85, and 95% RH was considerably lower at corresponding temperatures. The infectivity of larvae of P. odocoilei that survived desiccation was greatly reduced. Repeated freezing or repeated desiccation resulted in reduced survival of P. odocoilei and P. tenuis. Larvae of Parelaphostrongylus odocoilei from mule deer of Jasper National Park, Canada, were better able to resist the effects of freezing, but less able to resist the effects of desiccation, than were larvae of P. tenuis from white-tailed deer of Pennsylvania, USA.

Effects of source of metacercariae on experimental infection of Zygocotyle lunata (Digenea: Paramphistomidae) in CD-1 mice.

Outbred (CD-1 strain) mice were infected experimentally with metacercariae of Zygocotyle lunata obtained from naturally infected Helisoma trivolvis. Growth, survival, and fecundity of the parasite and gross pathology in the host were evaluated. Metacercariae from 5 snail sources produced 3 patterns of growth and survival: significantly smaller worms that survived less than 4 wk and did not mature, significantly larger worms that matured but survived less than 6 wk, and worms of intermediate size that matured and lived at least 19 wk. Parasite maturation (at 3-4 wk) and egg production (14 eggs.worm-1.hr-1) were similar among infections from different sources. Ceca of mice with infections at least 4 wk old were enlarged with localized mucosal alteration at the site of parasite attachment, but there was no evidence of splenomegaly. The differences in infection due to the source of metacercariae within 1 strain of final host were similar to differences reported among other species of final host. These results suggest that variation associated with the intermediate host may confound studies of specificity to the definitive host.

Parasitology meets ecology on its own terms: Margolis et al. revisited.

We consider 27 population and community terms used frequently by parasitologists when describing the ecology of parasites. We provide suggestions for various terms in an attempt to foster consistent use and to make terms used in parasite ecology easier to interpret for those who study free-living organisms. We suggest strongly that authors, whether they agree or disagree with us, provide complete and unambiguous definitions for all parameters of their studies.