RNAs in the sera of Persian Gulf War veterans have segments homologous to chromosome 22q11.2.

Reverse transcriptase PCR (RT-PCR) was used for polyribonucleotide assays with sera from deployed Persian Gulf War veterans with the Gulf War Syndrome and a cohort of nonmilitary controls. Sera from veterans contained polyribonucleotides (amplicons) that were obtained by RT-PCR and that ranged in size from 200 to ca. 2,000 bp. Sera from controls did not contain amplicons larger than 450 bp. DNA sequences were derived from two amplicons unique to veterans. These amplicons, which were 414 and 759 nucleotides, were unrelated to each other or to any sequence in gene bank databases. The amplicons contained short segments that were homologous to regions of chromosome 22q11.2, an antigen-responsive hot spot for genetic rearrangements. Many of these short amplicon segments occurred near, between, or in chromosome 22q11.2 Alu sequences. These results suggest that genetic alterations in the 22q11.2 region, possibly induced by exposures to environmental genotoxins during the Persian Gulf War, may have played a role in the pathogenesis of the Gulf War Syndrome. However, the data did not exclude the possibility that other chromosomes also may have been involved. Nonetheless, the detection of polyribonucleotides such as those reported here may have application to the laboratory diagnosis of chronic diseases that have a multifactorial etiology.

Cyclical micromovement and fracture healing.

We studied 56 patients with fractures of the tibial shaft in a multicentre prospective randomised trial of three methods of external fixation. Group I was treated with a fixator which was unlocked at 4 to 6 weeks to allow free axial compression (axial dynamisation) with weight-bearing. Group II was treated with a fixator that was similarly unlocked at 4 to 6 weeks but included a small silicone spring which on weight-bearing could be compressed by up to 2 mm. this spring returns to its original length on cessation of weight-bearing thus allowing cycles of motion of up to 2 mm. Group III had a spring fixator like group II, but it was unlocked from the start to allow cyclical micromovement as soon as weight-bearing began. Fracture healing was monitored by the measurement of fracture stiffness. We defined healing as achieving a stiffness of 15 Nm per degree. The mean time was 14.1 weeks in group I, 15.9 weeks in group II, and 19.3 weeks in group III. The difference between groups was statistically significant (p = 0.004). The 95% confidence intervals for the average delay in healing with early cyclical micromovement (group III) as compared with later axial dynamisation (group I) was 1.8 to 8.7 weeks. The healing time in patients whose cyclical micromovement was delayed for 4 to 6 weeks (group II) was between these two extremes, but the differences from either of the other groups could have been due to patient selection. In the patients who completed the full trial, there were pin-track infections in over 60% of those in the cyclical micromovement groups compared with 20% in the axial dynamisation group (p = 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)

A comparative study of early motion and immediate plaster splintage after internal fixation of unstable fractures of the ankle.

We reviewed 47 patients following operatively treated ankle fracture-dislocation, at an average of 15 months after injury, to assess the outcome of two different postoperative regimens. Of the 47 patients, 27 received early active and passive ankle exercises, and 20 patients received immediate plaster splintage. Patients were assessed clinically by an independent surgeon and subjective, objective and radiological criteria recorded. No significant difference was apparent between the two groups on any of the criteria, although the early movement group contained more patients who were completely pain free, had a normal gait and no radiological signs of arthrosis (P < 0.05). This was achieved at the expense of a longer stay in hospital (average 10.2 days versus 7.4 days for plaster splintage) and more ankle swelling.

Toxicity of Fusarium moniliforme var. subglutinans for chicks, ducklings, and turkey poults.

Corn-based diets contaminated with various concentrations of a moniliformin-producing isolate of Fusarium moniliforme var. subglutinans were found to be lethal for chicks, ducklings, and turkey poults. Ducklings appeared to be the most sensitive to the lethal effects of the toxic feed. Gross lesions were ascites, hydropericardium, and myocardial pallor. Microscopic lesions were limited to the heart and liver, and they consisted of degeneration and necrosis of the myocardium and degeneration of hepatocytes. Cardiotoxicosis was the apparent cause of death.

Citrinin mycotoxicosis in the rabbit: ultrastructural alterations.

Citrinin was given to rabbits as a single oral dose of 120 or 67 mg/kg. Rabbits were killed at 4, 6, 8, 10, and 12 hours post dosing, and the kidneys were fixed by intravascular perfusion. Ultrastructural alterations were evident by 4 hours after treatment. In the proximal tubule, alterations were brush border disruption, cytoplasmic rarefaction, and swelling of interdigitating processes. At higher doses, mitochondria were condensed and distorted. Medullary and straight cortical distal tubules had marked distention of the intercellular spaces and disorganization of interdigitating processes. Changes in cortical and outer medullary collecting ducts were similar but less severe. Renal alterations were suggestive of damage to membrane structure and/or transport functions and interference with cellular bioenergetics. Leukocytic infiltration was associated with damaged tubules indicating a contribution of inflammation to the development of the lesions.

Natural occurrence of deoxynivalenol, 15-acetyl-deoxynivalenol, and zearalenone in refusal factor corn stored since 1972.

Two samples of "refusal factor" corn, one stored frozen in Minnesota and one stored dry in Indiana since 1972 or 1973, were analyzed for the presence of Fusarium spp. and Fusarium toxins. Both samples were from corn refused by swine in Indiana from 1972 to 1973. Sample FS 808 (stored in Indiana) contained 20 ppm of deoxynivalenol (20 micrograms/g), 16 ppm of 15-acetyl-deoxynivalenol, 5 ppm of zearalenone, and 0.2 ppm of alpha-zearalenol. Sample FS 362 (stored in Minnesota) contained 3 ppm of deoxynivalenol, 1 ppm of 15-acetyldeoxynivalenol, and 0.3 ppm of zearalenone. The presence of 15-acetyl-deoxynivalenol is significant because it is the first report of it occurring naturally in refusal factor corn, and it may account in part for the refusal that could not be solely attributed to deoxynivalenol.

Citrinin mycotoxicosis in the rabbit: clinicopathological alterations.

Citrinin, a nephrotoxic mycotoxin, was dissolved in 0.5 N-NaOH neutralized with HCl and given in a single oral dose of 120 mg/kg (Trial I) or 80 or 100 mg/kg (Trial II) to male New Zealand white rabbits weighing 2.0-2.7 kg. In Trial I, sequential measurements of clinicopathological parameters were made over a 24-hr period. Azotaemia and metabolic acidosis with haemoconcentration and hypokalaemia developed within 4-12 hr. In Trial II, clinicopathological and urinary parameters were measured daily for 7 days. Increased blood urea nitrogen and serum-creatine levels and decreased creatinine clearance indicated renal failure; these values were most abnormal on days 2-4, returning to normal or near normal by day 7 in rabbits that survived. Urine analysis indicated tubular dysfunction and necrosis with glucosuria, isosthenuria and cylindruria; most urinary parameters were normal by day 7.

Studies on the sequential development and pathogenesis of citrinin mycotoxicosis in turkeys and ducklings.

The toxic effects of citrinin in turkeys and ducklings was studied in four trials. Citrinin dissolved in dimethyl sulfoxide-70% ethanol solution (3:1, volume/volume) was administered by gavage to male turkey poults and male white Pekin ducklings. When seven-day-old ducklings were given doses of citrinin between 30 to 110 mg/kg body weight, most of the treated ducklings which died (49/80) did so within four to 12 hours. Blood samples were collected sequentially at 3, 6, 12, and 24 hours after administration from seven-day-old ducklings given the single lethal dose (LD50). The alterations included hyperkalemia (P less than or equal to 0.01) and metabolic acidosis characterized by reduced blood pH (P less than or equal to 0.01) and base excess (P less than or equal to 0.01). Fourteen-day-old turkeys and ducklings given 56 or 57 mg/kg, respectively, were killed at 1, 3, 6, 12, 24, 48, and 72 hours after treatment. The principal alteration in both species was nephrosis that was more severe in turkeys than in ducklings. Tubular necrosis was the dominant lesion at three to 72 hours in turkeys and at six to 24 hours in ducklings. Hepatic and lymphoid lesions occurred in both turkeys and ducklings treated with citrinin.

Mycotoxicoses produced in ducklings and turkeys by dietary and multiple doses of citrinin.

Citrinin was mixed in the diet (100, 250, 500 parts/10(6)) and fed to 1-day-old male White Pekin ducklings (Trial I), or it was dissolved in dimethyl-sulphoxide-70% ethanol (3:1, v/v) and administered by crop gavage to 14-day-old male White Pekin ducklings (Trial II) or 7-day-old male turkey poults (Trial III). Ducklings given 250 or 500 parts/10(6) citrinin diet for 15 days had clinical signs of citrinin toxicity. Body weight gain was significantly decreased by the 500 parts/10(6) diet (P < 0.05). Ducklings fed 250 and 500 parts/10(6) developed nephropathy that was more severe in the 500 parts/10(6) group. The nephropathy was characterised by degeneration, necrosis, mineralisation and regeneration of tubular epithelial cells of both the cortical and medullary regions. Interstitial fibrosis was found in the medullary regions of the 500 parts/10(6) group only and focally was accompanied by interstitial nephritis. Seven daily doses of citrinin equal to a half or three-quarters of the single-dose LD(50) produced no additive toxic effects in either ducklings or turkeys, but body weight gain was significantly decreased in duckling given three-quarters of the single-dose LD(50).

Citrinin mycotoxicosis in the rabbit.

In three trials, single or multiple doses of citrinin dissolved in 0.5 N-NaOH and adjusted to neutral pH with HCl were given to rabbits by either the oral or intraperitoneal route. The 72-hr LD50 was 50 mg/kg body weight by intraperitoneal administration and 134 mg/kg by the oral route. The primary clinical sign in rabbits receiving a single oral dose of 125-150 mg citrinin/kg was fluid diarrhoea commencing 8 hr after dosing. Pathological alterations were generally confined to the kidney and consisted of degeneration and necrosis of proximal convoluted tubules and straight segments. In rabbits given a single oral dose of citrinin (130 mg/kg) the earliest histopathological change, seen 8 hr after dosing, was cytoplasmic vacuolation of tubular epithelial cells. Rabbits given a single oral dose of 120 mg citrinin/kg had regeneration of renal tubular epithelium accompanied by slight tubular cell necrosis when examined 7 days after dosing. Rabbits given multiple sublethal doses of citrinin (33.5 or 77 mg/kg daily for 7 days) had renal alterations of mild tubular degeneration and necrosis, and tubular regeneration.

Effect of Fusarium roseum (Gibberella zea) on pregnancy and the estrous cycle in gilts fed molded corn on days 7-17 post-estrus.

A laboratory produced corn culture of Fusarium roseum (Gibberella zea) was fed as 10% of the total diet to bred and non-bred gilts on days 7-17 postestrus. The corn culture contained both zearalenone and deoxynivalenol by analysis, and caused considerable feed refusal. Although 6 of 10 bred gilts fed the corn culture appeared to be pregnant by ultrasound testing between days 40-50 post-breeding, none of them farrowed. Progesterone analyses indicated that none of the 10 gilts cycled normally during the 20 week observation period. Three maintained high serum progesterone (greater than 10 ng/ml) during the entire period. Progesterone analyses were similar in 5 non-bred gilts fed the corn culture in that none of them cycled normally in 20 weeks. The retention or loss of corpora lutea could not be attributed to levels of luteinizing hormone, as no relationship was found between concentrations of progesterone and luteinizing hormone. This experiment indicates that limited dietary exposure of female swine to Fusarium roseum infected corn can result in embryonic loss and disruption of normal reproductive cycling for an extended length of time.

Acute toxicity of citrinin in turkeys and ducklings.

Citrinin, a naturally occurring mycotoxin, was dissolved in dimethyl-sulphoxide - 70% ethanol (3:1, v/v) and administered orally in two trials to 7-day-old male turkey poults and male white Pekin ducklings. The single dose LD50 value in 7-day-old male turkey poults was 56 mg/kg and in 7-day-old male white Pekin ducklings it was 57 mg/kg. The mycotoxin was nephrotoxic in both species, but the renal lesions were more severe in turkeys and were characterised by degeneration and necrosis of renal tubular epithelium. In turkeys, lesions were found in the liver and included hepatic cell necrosis and biliary hyperplasia. Lymphoid necrosis with depletion involved the thymus and cloacal bursa of turkeys and ducklings. These latter lesions were the most prominent histopathological alterations in citrinin-treated ducklings.

Effect of Fusarium roseum corn culture containing zearalenone on early pregnancy in swine.

A corn culture of Fusarium roseum was added to a standard corn-soybean swine gestation ration. Low, middle, and high dosage mixed feeds contained 7, 38, and 64 mg of zearalenone/kg of feed (7, 38, and 64 ppm) and 0.5, 2.5, and 4.5 mg of deoxynivalenol/kg, respectively. Control feed was the standard ration without added F roseum corn culture. Mature gilts were bred by natural service and fed control or F roseum molded feed from 3 to 34 days after breeding. The main effect of the molded feed was an inhibition of fetal development, with decreased numbers of fetuses present in treated animals at slaughter (38 to 43 days after breeding). Normal litters were present in 7 of 8 control animals, in 2 of 4 gilts given the low-dosage feed, in 1 of 4 gilts given the medium dosage, and in 0 of 4 given the high-dosage feed. Corpora lutea were maintained in all treated animals, as evidenced by serum progesterone concentrations. Serum estradiol concentrations were decreased in gilts in the middle- and high-dosage groups. The genital system of the gilts fed low- and middle-dosage feeds had a gross and microscopic appearance similar to that of the pregnant controls and reflected prolonged progesterone stimulation. Morphologic changes in the genital system of the high-dosage group were intermediate between changes induced by progesterone and those induced by estrogen. Clinical signs of hyperestrogenism and partial feed refusal were noticed in only some of the high-dosage group animals.

Experimental trichothecene mycotoxicosis produced in broiler chickens by Fusarium sporotrichiella var. sporotrichioides.

Fusarium sporotrichiella var. sporotrichioides (Bilay), cultured on . sterilised popcorn at 23 degrees C and then at 8 degrees C, 16 degrees C and 23 degrees C and fed as 50% of the diet, was lethal to 7-day-old male broiler chickens. The 8 degrees C culture, containing T-2 toxin at 50 parts per million (ppm) and neosolaniol at 5 ppm, was given as whole culture at dietary concentrations of 10%, 5%, 1% and 0% for 17 days and 1% for 42 days. Half the chickens that were fed the 10% diet died during the 17 days (5 ppm T-2 toxin and 0.5 ppm neosolaniol). The corresponding daily dose was 0.24 mg T-2 toxin and 0.02 mg neosolaniol/kg body weight/day. The chickens that died were dehydrated, had necrosis and depletion of lymphoid and haematopoietic tissues and necrosis of the hepatobiliary system, gastroenteric mucosa, feather epidermis and renal tubular epithelium. The survivors had anaemia, reduction of weight gain and transiently altered righting reflex. The comb and beak were pale yellow and the feather barbs were dishevelled. Survivors also had atrophied lymphoid tissues, reduced haematopoietic cellularity in the bone marrow, necrosis of oral and crop mucosa, vacuolated hepatocytes, hyperplastic bile ductules, and reduction of the thyroid follicular diameter.

Influence of prepubertal consumption of zearalenone on sexual development of boars.

Littermate pairs of crossbred boars were fed 40 ppm zearalenone from 14 to 18 weeks of age, and the effects on reproductive development were examined. Compared to controls, boars fed zearalenone subsequently had reduced libido scores (P greater than.05), and fewer treated animals exhibited mating behavior. Plasma concentrations of testosterone were depressed (P greater than .05) during consumption of zearalenone. Zearalenone did not significantly affect age at which sperm were first voided in the urine (P greater than .25). Body weight and testicular size were not affected by zearalenone. At 36 weeks of age, testicular weight, epididymal weight, spermatid numbers, motility of cauda epididymal sperm and testicular morphology did not differ between control boars and those that had consumed zearalenone.

Mycotoxicosis produced in swine by cultural products of an isolate of Aspergillus ochraceus. II. Clinicopathologic changes.

Rice culture of a toxigenic strain of Aspergillus ochraceus was fed at a concentration of 25% to weanling pigs for 10 days. The clinicopathological abnormalities reflected renal damage. Activities of lactic dehydrogenase, glutamic-oxaloacetic transaminase and isocitrate dehydrogenase were increased in the urine but not in the serum. Serum concentrations of urea nitrogen and creatinine were high. Cellular and granular casts, blood, protein, and glucose were in the urine of pigs fed toxic diet. Serum concentrations of K+, Na+ and Cl- were unchanged, but concentrations of these electrolytes were reduced in the urine.

Mycotoxicosis produced in swine by cultural products of an isolate of Aspergillus ochraceus. I. Clinical observations and pathology.

Pigs fed a ration, 25% of which was rice culture, of Aspergillus ochraceus lost weight or failed to gain and became depressed. Some pigs died and most developed subcutaneous edema, hydrothorax, hydroperitoneum, pulmonary atelectasis, edema of the mesentery and perirenal edema. Microscopic lesions in addition to edema were primarily renal and consisted of tubular degeneration and necrosis, hyaline tubular casts, interstitial fibrosis and tubular cell regeneration. The first change found after 3 days was cytoplasmic vacuolation of the convoluted and straight segments of the proximal tubules. Necrotic proximal tubules were found after 4 days and after 9 days degeneration and necrosis involved predominantly proximal tubular segments. Pigs fed a ration, 12.5% of which was rice culture, for 8 weeks did not develop perirenal edema but had firm kidneys. Extensive interstitial fibrosis of the cortical labyrinth was the principal change. Within the fibrous connective tissue, some tubules were necrotic and others were atrophied.