Biomedical subjects

D M Creasy

Publications and source records attributed to D M Creasy.

31 records · Page 2Linked to original sources

PubMed1985

A quantitative study of stage-specific spermatocyte damage following administration of ethylene glycol monomethyl ether in the rat.

A quantitative study has been carried out to characterize the stage susceptibility of the spermatocyte to ethylene glycol monomethyl ether (EGM) toxicity. EGM was administered as a single oral dose of 250 mg/kg body wt and rats were examined at time periods after dosing. The number of spermatocytes and round spermatids in tubules at each stage of spermatogenesis was counted. A sharp transition in susceptibility was observed between zygotene spermatocytes in stage XIV which showed no effect and pachytene spermatocytes in stage I which showed death or depletion of 70% of its population after 1 day. A similar transition was seen between dividing spermatocytes and step 1 spermatids, the latter being unaffected. There was a gradual reduction in susceptibility toward midpachytene such that cells in stages VII-XI showed no effect. Analysis of later time periods revealed no effect on spermatogonia or prepachytene spermatocytes but did indicate that midpachytene spermatocytes underwent delayed cell death after further progression through the cycle. In a separate sequential morphological study of early changes, the earliest signs of necrosis were seen 12 hr after dosing and were restricted to spermatocytes in stages V, XI, and XII. Cell death then progressed in a wave-like manner through stages XIII and XIV finally reaching stage I, 24 hr after dosing.

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PubMed1985

The role of metabolism in 2-methoxyethanol-induced testicular toxicity.

The role of metabolism in 2-methoxyethanol (ME)-induced testicular toxicity has been investigated with Sprague-Dawley rats. Following administration of [14C]ME (250 mg/kg, ip) to a group of animals, there was evidence of testicular damage, identified as depletion of the spermatocyte population. Radioactivity detected in urine over 48 hr after treatment accounted for 55% of the dose. The major urinary metabolites were identified by HPLC and isotope dilution analysis, as methoxyacetic acid (MAA) and methoxyacetylglycine (accounting for 50 to 60% and 18 to 25%, respectively, of urinary radioactivity). Analysis of plasma revealed a rapid conversion of ME to MAA (t1/2 for disappearance of ME = 0.6 +/- 0.03 hr) and gradual clearance of radioactivity (t1/2 = 19.7 +/- 2.3 hr). Pretreatment of animals with pyrazole (400 mg/kg, ip) 1 hr prior to [14C]ME dosing gave complete protection against the testicular toxicity of ME. Radioactivity detected in the urine from the pyrazole-pretreated groups over 48 hr (18%) was significantly lower than in the ME-only group. The major radioactive peak co-chromatographed with ME (30 to 36% of the total urinary radioactivity). MAA and methoxyacetylglycine were not major metabolites. Analysis of plasma revealed almost complete inhibition of the conversion of ME to MAA (t1/2 for disappearance of ME = 42.6 +/- 5.6 hr, clearance of radioactivity t1/2 = 51.0 +/- 7.8 hr). The results demonstrate that metabolic activation is required for 2-methoxyethanol to exert toxicity to the male reproductive system.

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PubMed1985

Studies on the toxicity of some glycol ethers and alkoxyacetic acids in primary testicular cell cultures.

Primary mixed cultures of Sertoli and germ cells were prepared from testes of immature rats and their response to the known testicular toxicants ethylene glycol monomethyl ether (EGM) and ethylene glycol monoethyl ether (EGE) was studied. Neither EGM nor EGE produced any morphological evidence of toxicity when added to the culture medium at up to 50 mM for 72 hr. In contrast, their metabolites methoxyacetic acid (MAA) and ethoxyacetic acid (EAA) at 2 to 10 mM for 24 to 72 hr caused degeneration of the pachytene and dividing spermatocytes, the target cells of the parent ethers in vivo. As in vivo, earlier spermatocytes, spermatogonia, and Sertoli cells appeared unaffected. EAA was less potent than MAA whereas n-propoxy- and n-butoxyacetic acid, and methoxyacetylglycine, a further metabolite of MAA, produced no morphological changes under these conditions. The same order of toxicity was observed in concurrent studies with the four acids in rats. In culture, the severity of the morphological changes was paralleled by decreases in the activity of carnitine acetyltransferase and lactate dehydrogenase-X in the attached germ cell fraction. Analysis of culture medium provided no evidence for the conversion of EGM to MAA or other metabolites or for the further metabolism of MAA. The close correspondence between the testicular toxicity of alkoxyacetic acids in culture and in vivo suggests a similar mode of action in both cases and points to the potential value of these cultures for mechanistic studies and for screening purposes. The results also emphasize the role of metabolism in the testicular toxicity of glycol ethers and indicate that MAA is an active metabolite of EGM.

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PubMed1984

The morphological development of glycol ether-induced testicular atrophy in the rat.

The testicular effects of daily oral dosing with two glycol ethers--either ethylene glycol monomethyl ether (EGM) or monoethyl ether (EGE) were studied in the prepubertal rat by histological examination of the testes. Over the 11-day dosing period studied, EGM was found to produce testicular damage at dose levels of and in excess of 100 mg/kg/day with a no-effect level at 250 mg/kg/day. The findings at sequential time intervals throughout the dosing period indicated that primary spermatocytes undergoing pachytene development constituted the initial and major site of morphological damage. Within this population, differential sensitivity was demonstrated depending on the precise stage of meiotic maturation. A consistent order of spermatocyte susceptibility emerged from the results: dividing spermatocytes (Stage XIV) greater than early-pachytene spermatocytes (Stages I-III) greater than late-pachytene spermatocytes (Stages IX-XIII) greater than midpachytene spermatocytes (Stages IV-VIII). Leptotene/zygotene spermatocytes and Step 1 spermatids also showed degenerative changes but only after prolonged dosing at high-dose levels. The significance of the findings with respect to mechanisms of cellular toxicity in the testis is discussed.

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PubMed1984

Testicular toxicity produced by ethylene glycol monomethyl and monoethyl ethers in the rat.

Ethylene glycol monomethyl ether (EGME) and ethylene glycol monoethyl ether (EGEE) were administered orally to young male rats at doses varying from 50 to 500 mg/kg/day and 250 to 1000 mg/kg/day for EGME and EGEE, respectively, for 11 days. At sequential times animals were killed and testicular histology examined. The initial and major site of damage following EGME treatment was restricted to the primary spermatocytes undergoing postzygotene meiotic maturation and division. EGEE produced damage of an identical nature, but a larger dose was required to elicit equivalent severity (500 mg EGEE/kg being approximately equivalent to 100 mg EGME/kg). Additionally, within the spermatocyte population, differential sensitivity was observed depending on the precise stage of meiotic maturation: dividing (stage XIV) and early pachytene (stages I-II) greater than late pachytene (stages VIII-XIII) greater than mid-pachytene (stages III-VII). Equivalent doses of methoxyacetic acid (MAA) and ethoxyacetic acid (EAA) gave injury similar to the corresponding glycol ether. When animals were pretreated with inhibitors of alcohol metabolism followed by a testicular toxic dose of EGME (500 mg/kg), an inhibitor of alcohol dehydrogenase (pyrazole) offered complete protection. Pretreatment with the aldehyde dehydrogenase inhibitors disulfiram or pargyline did not ameliorate the testicular toxicity of EGME. In mixed cultures of Sertoli-germ cells, MAA and not EGME produced effects on spermatocytes analogous to that seen in vivo, at concentrations approximately equivalent to steady-state plasma levels after a single oral dose of EGME (500 mg/kg). It would seem likely that a metabolite (MAA or possibly methoxyacetaldehyde) and not EGME is responsible for the production of testicular damage.

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PubMed1983

The morphological development of di-N-pentyl phthalate induced testicular atrophy in the rat.

Prepubertal rats treated orally with di-n-pentyl phthalate at 2.2 g/kg body weight were killed at 1, 3, 6 and 24 hr following a single dose, and after 2, 3 and 4 days of repeated daily dosing. At 3 hr Sertoli cells in a proportion of the seminiferous tubules showed vacuolation of the perinuclear smooth endoplasmic reticulum with an associated inward displacement of germinal cells. By 6 hr the vacuolation had extended to the apical cytoplasm and was evident in most tubules. Early degenerative changes were also apparent in spermatocytes and spermatids and were accompanied by an acute interstitial inflammatory infiltrate. By 24 hr, germinal cell degeneration was extensive with desquamation and general disorganisation of cell layers within the epithelium, but the interstitial inflammatory infiltrate had declined. Mitochondrial succinic dehydrogenase activity in Sertoli cells was reduced at 3 and 6 hr and absent by 24 hr. In germinal cells it was unaffected at 3 and 6 hr but absent by 24 hr. Two, three and four days of daily phthalate treatment resulted in a gradual depletion of germinal cells from all tubules, leaving a Sertoli cell matrix containing a few necrotic spermatocytes and occasional normal spermatogonia. The significance of the early Sertoli cell changes is discussed.

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PubMed1983

Testicular toxicity of ethylene glycol monomethyl and monoethyl ethers in the rat.

Ethylene glycol monomethyl (EGM) and monoethyl (EGE) ethers were administered po to rats at dosages varying from 50 to 500 mg/kg body weight/day for EGM and 250 to 1000 mg/kg body weight/day EGE for 11 days. First evidence of testicular damage following EGM treatment was observed 24 hr after a single dose of 100 mg/kg body weight when the lesion appeared localized in the primary spermatocyte. At 16 hr after a single dose of 500 mg/kg, mitochondrial damage was one of the first subcellular changes to be demonstrated. Treatment of animals with EGE resulted in a similar lesion; however, to obtain damage of equivalent severity, a larger dosage for a longer period was required. In limited studies with 2-methoxy- and 2-ethoxyacetic acids (putative metabolites of EGM and EGE, respectively), using equimolar doses to their parent compounds (500 mg EGM or EGE/kg for 4 or 11 days, respectively) gave damage of equivalent severity to the corresponding glycol ether. After dosing animals with 500 mg EGM/kg body weight for 4 days, the testes recovered weight, and the majority of tubules recovered their spermatogenic potential within one full maturation cycle. The recovery study also indicated a possible effect on the spermatogonia in a small number of tubules although no morphological abnormalities to this cell type could be observed. No effect levels over the 11-day treatment period were 50 and 250 mg/kg body weight/day for EGM and EGE, respectively.

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PubMed1979

Prostacyclin (PGI2) effects on anterior pituitary hormones in the rat in vivo.

Intravenous injection of 600 microgram PGE2 or PGI2 significantly increased serum LH and prolactin levels in estradiol treated ovariectomized rats. There was no effect on serum FSH concentration. PGE2 and PGI2 stimulated LH release in a non-dose dependent manner, while prolactin levels were positively correlated with the dose administered following PGI2 treatment. 6-keto-PGF1 alpha at a comparable dose had no effect on pituitary hormone levels. Subcutaneous administration of 1 mg/kg or 60 mg/kg PGI2 for seven days significantly depressed serum LH level both in male and female rats. These doses had no effect on serum FSH or prolactin levels.

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PubMed1976

Wholebody autoradiography: a beta-radioluminescence method for detecting tritium.

A method of increasing the sensitivity of X-ray film for the wholebody autoradiography of tritiated compounds is described. Thirty micrometre sections are sprayed with a scintillator, Diphenyl oxazole (PPO) and exposed to light sensitive (screen) X-ray film at -70 degrees C. The method permits a substantial reduction in the required dose of tritium and of the period of exposure of tissue sections.

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PubMedDate not supplied

Two-generation reproduction toxicity studies of di-(C(7)-C(9) alkyl) phthalate and di-(C(9)-C(11) alkyl) phthalate in the rat.

Di-(C(7)-C(9) alkyl) phthalate (D79P) and di-(C(9)-C(11) alkyl) phthalate (D911P), based on high-normality linear oxo-alcohols, have been assessed for their impact upon reproductive performance in Sprague-Dawley rats. Rats were continuously exposed to either D79P or D911P at dietary levels of 0%, 0.1%, 0.5%, or 1.0% over two generations. Selected F(0) offspring (F(1) generation) were exposed to the same dietary concentration of D79P or D911P as the respective F(0) animals, and were mated to produce F(1) offspring. Both D79P and D911P markedly reduced body weight gain in F(0) and F(1) adult males at the highest dose, but females were affected to a lesser extent. There was no impairment of fertility, fecundity, or development in either generation, but body weights of offspring in the 1.0% D79P and 1.0% D911P groups were slightly and transiently reduced over the weaning period. Although decreases in the weight of several organs were accounted for by depressed body weight, ovary weights were reduced in both generations exposed to 1.0% D79P, and epididymidal weights were slightly reduced in adults of both generations exposed to 1.0% D911P. However, ovarian function-assessed by the oestrus cycle and mating behaviour-and epididymidal sperm concentration, motility, and morphology were unaffected by either substance. Treatment resulted in liver changes, particularly in males, characterised by increased liver weight in young animals, histopathologic changes and reduced organ weight in mature animals, and an increase in palmitoyl CoA oxidase activity. In conclusion, neither D79P nor D911P impaired reproductive function in rats when administered in the diet at levels that induce systemic toxicity, and the NOAEL for effects on reproduction in the rat is 0.5% for both D79P and D911P.

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PubMedDate not supplied

Pathogenesis of male reproductive toxicity.

Toxicologic disturbance of male reproductive function can occur at many sites and produce a range of effects, some primary and some secondary to the initial insult. The challenge to the toxicological pathologist is to identify the primary site of damage and provide an insight into the pathogenesis of the morphologic lesion or functional deficit. Target sites include the testis, the epididymis, the mature sperm, and the hormonal regulatory system. Detection of effects at these varied sites requires the measurement of multiple endpoints only 1 of which is histopathology, but once identified, careful microscopic examination of the early changes in lesion development can provide essential information on the probable target cell and possible mechanisms of toxicity. Chemicals that affect different cell types or specific cellular functions generally elicit predictable patterns of pathological changes that can be readily recognized. Understanding the pathogenesis, the likely reversibility and the significance of reproductive tract lesions is aided by a sound knowledge of the physiology of the testis and epididymis and, in particular, an understanding of the timing of sperm production and transport.

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PubMedDate not supplied

Oral (drinking water) two-generation reproductive toxicity study of dibromoacetic acid (DBA) in rats.

In a two-generation study of dibromoacetic acid (DBA), Crl SD rats (30 rats/sex/group/generation) were provided DBA in drinking water at 0 (reverse osmosis-deionized water), 50, 250, and 650 ppm (0, 4.4 to 11.6, 22.4 to 55.6, and 52.4 to 132.0 mg/kg/day, respectively; human intake approximates 0.1 microg/kg/day [0.0001 mg/kg/day]). Observations included viability, clinical signs, water and feed consumption, body and organ weights, histopathology, and reproductive parameters (mating, fertility, abortions, premature deliveries, durations of gestation, litter sizes, sex ratios and viabilities, maternal behaviors, reproductive organ weights, sperm parameters and implantation sites, sexual maturation). Histopathological evaluations were performed on at least 10 P and F1 rats/sex at 0 and 650 ppm (gross lesions, testes, intact epididymis; 10 F1 dams at 0, 250, and 650 ppm for primordial follicles). Developmental observations included implantations, pup numbers, sexes, viabilities, body weights, morphology, and reproductive performance. At 50 ppm and higher, both sexes and generations had increased absolute and relative liver and kidneys weights, and female rats in both generations had reduced absolute and relative adrenal weights; adrenal changes were probably associated with physiological changes in water balance. The livers and kidneys (10/sex/group/generation) had no histopathological changes. Other minimal effects at 50 ppm were reduced water consumption and a transient reduction in body weight. At 250 and 650 ppm, DBA reduced parental water consumption, body weight gains, body weights, feed consumption, and pup body weights. P and F1 generation male rats at 250 and 650 ppm had altered sperm production (retained step 19 spermatids in stages IX and X tubules sometimes associated with residual bodies) and some epididymal tubule changes (increased amounts of exfoliated spermatogenic cells/residual bodies in epididymal tubules, atrophy, and hypospermia), although inconsistently and at much lower incidences. Unilateral abnormalities of the epididymis (small or absent epididymis) at 650 ppm in four F1 generation male rats were considered reproductive tract malformations. The no-observable-adverse-effect level (NOAEL) and reproductive and developmental NOAELs for DBA were at least 50 ppm (4.5 to 11.6 mg/kg/day), 45,000 to 116,000 times the human adult exposure level. Reproductive and developmental effects did not occur in female rats exposed to DBA concentrations as high as 650 ppm. Based on the high multiples of human exposure required to produce effects in male rats, DBA should not be identified as a human reproductive or developmental risk.

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PubMedDate not supplied

Evaluation of testicular toxicity in safety evaluation studies: the appropriate use of spermatogenic staging.

Toxicology of the male reproductive system has received increased interest in recent years partly fuelled by the growing reports of falling sperm counts and rising reproductive disorders in the human population. Recently revised regulatory guidelines for the safety assessment of pharmaceuticals and chemicals on reproduction and fertility have emphasized the importance of detailed histopathological examination of the testes as a sensitive method for detecting disturbances in spermatogenesis. Unfortunately this has been accompanied by a general confusion regarding a practical approach to undertaking such a detailed examination, particularly in respect to the use of spermatogenic or tubular staging to identify subtle disturbances in spermatogenesis. The ability to identify tubular stages of the spermatogenic cycle in sections of testis plus a good understanding of the spermatogenic process and its dynamics are essential in order to carry out a sensitive of testicular histopathology and to interpret the changes seen. A rational approach is required initially to detect and subsequently to characterize toxic effects to the male reproductive system. It is important that a distinction is made between these two objectives since different study designs are required and different methodology may be employed to produce the type of information or data required.

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