High Levels Of Acid Phosphatase Research Articles

Successful treatment of Gaucher disease type 1 by enzyme replacement therapy over a 10-year duration in a Japanese pediatric patient: A case report.

The prevalence of Gaucher disease (GD) in Japan is much lower than that in Western countries; therefore, data on Japanese pediatric patients with GD type 1 are currently limited. The present study reports on the case of a Japanese pediatric patient with GD type 1 who was diagnosed when she presented with hepatosplenomegaly, thrombocytopenia and slight anemia at the age of 2 years. Serology tests revealed high levels of acid phosphatase (ACP) and angiotensin-converting enzyme (ACE). A bone marrow biopsy revealed the presence of Gaucher cells. Abdominal MRI indicated huge hepatosplenomegaly. Erlenmeyer flask deformity was observed on X-ray examination. MRI of the femora featured a high-intensity area within the diaphysis region. The enzymatic activity of leukocyte β-glucosidase, the measurement of which is necessary for a definitive diagnosis of GD, had decreased to 186.7 nmol/h/mg (reference range, 1,424.0-2,338.0 nmol/h/mg). Based on these results, the patient was clinically diagnosed with GD. Glucocerebrosidase gene analysis identified the compound heterozygote mutation of F213I (c.754T>A) on exon 7 and L444P (c.1448T>C) on exon 11. Enzyme replacement therapy (ERT) along with an intravenous infusion of 60 U/kg of imiglucerase every other week was initiated following diagnosis. Hemoglobin levels and the platelet count gradually improved and normalized after two years. ACP and ACE levels, biomarkers of the progression of GD, also improved. Abdominal MRI at six months after the initiation of ERT revealed a decrease in the size of the liver and spleen, which normalized after 1 year. Conversely, MRI of the femora indicated no improvement in the high-intensity area within the diaphysis region for 10 years.

КОМПЛЕКСНЕ ЛІКУВАННЯ СИНДРОМУ ДІАБЕТИЧНОЇ СТОПИ

The aim was to develop a comprehensive treatment of DFS, using reconstructive surgeries and indirect revascularization methods. Our work is based on 25 year experience of treatment of 1107 patients with DFS, who were treated in Ivano-Frankivsk Central hospital. Patients were examined on admission: common clinical methods, angiography, densitometry, electronic microscopy and histochemical examination of limb tissues, TcpO2 and regional cytokine reaction. Results showed that reconstructive operations on limb arteries, in case if it was possible, were the best method of treatment. Indications for surgery were TcpO2≤25 mm.hd. Mekenberg sclerosis III was contraindication for reconstructive surgery. Tissue vitality, its future regeneration possibility depends on succinate dehydrogenase and acid phosphatase. In vital mussels of the hip, shin and foot succinate dehydrogenase range was 3-5 points, in dead tissues- 0. Acid phosphatase in vital 0-1 points, in dead tissues- 4-5. Surgical tactic algorithm was developed ccording to electronic microscopy and histochemistry. It was based on tissue vitality and gave us an answer to what part of limb must be amputated. Indication for amputation was high level of acid phosphatase, bone tissue destruction, massive purulent destructions of soft tissues, gangrene. Bone trepanation causes decreasing of internal bone tissue, decreasing of intraossal receptor irritation, increases local microcirculation. For elimination of myelodepresion and erythropoiesis stimulation we offered intraspongious automyelotransplantation which caused to extra anatomical connection’s between intraossal and muscular neoangiogenesis. Topical treatment of trophic ulcers was performed by auto desmoplastic and cultures of fibroblasts which can cover defect as by themselves and as causes to better skin autotransplant healing. We get a positive effect of prolonged intraarterial antibiotic therapy with erytrocytal containers which make antibiotic exposition in infection localization up to 36 hours.

COMPREHENSIVE TREATMENT OF DIABETIC FOOT SYNDROME

The aim was to develop a comprehensive treatment of DFS, using reconstructive surgeries and indirect revascularization methods. Our work is based on 25 year experience of treatment of 1107 patients with DFS, who were treated in Ivano-Frankivsk Central hospital. Patients were examined on admission: common clinical methods, angiography, densitometry, electronic microscopy and histochemical examination of limb tissues, TcpO2 and regional cytokine reaction. Results showed that reconstructive operations on limb arteries, in case if it was possible, were the best method of treatment. Indications for surgery were TcpO2≤25 mm.hd. Mekenberg sclerosis III was contraindication for reconstructive surgery. Tissue vitality, its future regeneration possibility depends on succinate dehydrogenase and acid phosphatase. In vital mussels of the hip, shin and foot succinate dehydrogenase range was 3-5 points, in dead tissues- 0. Acid phosphatase in vital 0-1 points, in dead tissues- 4-5. Surgical tactic algorithm was developed ccording to electronic microscopy and histochemistry. It was based on tissue vitality and gave us an answer to what part of limb must be amputated. Indication for amputation was high level of acid phosphatase, bone tissue destruction, massive purulent destructions of soft tissues, gangrene. Bone trepanation causes decreasing of internal bone tissue, decreasing of intraossal receptor irritation, increases local microcirculation. For elimination of myelodepresion and erythropoiesis stimulation we offered intraspongious automyelotransplantation which caused to extra anatomical connection’s between intraossal and muscular neoangiogenesis. Topical treatment of trophic ulcers was performed by auto desmoplastic and cultures of fibroblasts which can cover defect as by themselves and as causes to better skin autotransplant healing. We get a positive effect of prolonged intraarterial antibiotic therapy with erytrocytal containers which make antibiotic exposition in infection localization up to 36 hours.

Hydroxyapatite Biosynthesis by a Serratia sp. and Application of Nanoscale Bio-HA in the Recovery of Strontium and Europium

ABSTRACTA Serratia sp. expresses a high level of acid phosphatase when grown continuously under carbon limitation. In the presence of CaCl2, biosynthesis of nanocrystalline hydroxyapatite (bio-HA) was achieved by utilizing phosphate released via enzymatic cleavage of an applied substrate (glycerol 2-phosphate: G2P). Hydroxyapatite crystals were identified by energy dispersive X-ray emission (EDX) and selected area diffraction (SAD). X-ray powder diffraction (XRD) analysis gave a mean crystallite size of ∼21–32 nm, with the smallest crystals (21–24 nm) obtained using 1 mM Ca2+ and 1 mM G2P. The uptake of Eu3+ and Sr2+ by bio-HA made by continuously pregrown cells (0.42 mg/mg and 0.043 mg/mg respectively) was ∼20% greater for Sr2+ than was previously reported for bio-HA material of size ∼40 nm made by batch-pregrown cells, while the corresponding uptake of Eu3+ was increased by > 1.8-fold. This was attributed to the localization of Eu (III) at grain boundaries by reference to previous work and highlights the potential of bio-HA as a sequestration agent for recovery of rare earth elements and trivalent actinides.

Comparative toxicity and biochemistry of organophosphates and pyrethroid compounds on both laboratory and field strain of the Cotton Leafworm Spodoptera littoralis (Boisd.)

Bioassay was carried out for monitoring resistance spectrum toward insecticides include organophosphates, Dursban H-48% EC (Chlorpyrifos) and pyrethroids Cyper (Cypermethrin 10% EC) in three different field populations (Gharbia , Kalubia and Menofeya Governorates) of the Cotton Leafworm, Spodoptera littoralis (Boisd.). Toxicity data based on LC50 values indicated that the pyrethroid Cyper (Cypermethrin 10% EC) is more toxic insecticide against the laboratory strain, while, the OP Dursban H- 48% EC (Chlorpyrifos) was less toxic. The development of resistance to organophosphates, Dursban H- 48% EC (Chlorpyrifos) and pyrethroids Cyper (Cypermethrin 10% EC) in field strains of the cotton leafworm, in relation to esterases, glutathione s- transferase, and phosphatases was studied. Field strains exhibited moderately high level of resistance to the tested insecticides. A significant increase in the activity of esterases, glutathione s- transferase suggests a good relation to resistance was detected. A high level of acid phosphatase and low level of alkaline phosphatase accompanied the level of resistance to pyrethroid and OP.

Replacement Therapy with Imiglucerase Improves Magnetic Resonance Imaging (MRI) Abnormalities in Almost Half Patients with Non-Neuronopathic Form of Gaucher Disease After Twelve Months of Therapy: Results of a Semi-Quantitative MRI Method.

Abstract 2132Gaucher disease (GD) is a lysosomal storage disease characterized by a genetic disruption in the metabolic breakdown of glucocerebroside caused by a lack of the enzyme beta-glucocerebrosidase. Non-neuronopathic GD form is accompanied by bony complications including osteoporosis, osteonecrosis, lytic lesions, pathological fractures and Erlenmeyer flask deformity. MRI is considered as the most sensitive imaging procedure for assessing skeletal involvement in GD patients. To-date, enzyme replacement therapy with imiglucerase is the standard of care for the treatment of patients with non-neuronopathic GD. Although this treatment has been used for many years, there is limited information for the effect of imiglucerase on bone deformities and MRI findings of GD patients.To address this issue, we studied 27 adult patients (15M/12F, median age 44.5 years) with non-neuronopathic GD who received replacement therapy with imiglucerase for at least one year. Bisphosphonates were not given in any patient during the study period. MRI of the femoral bones was performed in all patients before the initiation of replacement therapy and 12 months post-imiglucerase treatment. MRI of the femoral bones was also performed in 27, gender and age-matched healthy individuals, who served as controls. We implemented T1-weighted spin echo (T1W) sequences, proton density sequences with fat suppression (PDFS) to further evaluate the detected changes on T1W sequence and short time inversion recovery (STIR) sequences for the evaluation of disease activity. We also performed a thorough comparison of T1W and PDFS images targeting in the detection of the remodeling process of the affected bone. In all patients we evaluated bone infiltration on T1W images in the affected areas as opposed with those of healthy regions and in conjunction with measurements taken from subcutaneous fat of both thighs of the same patients. We followed exactly the same process throughout the whole reevaluation we performed one year after the initiation of replacement therapy with imiglucerase.The rate of subcutaneous fat of both healthy and affected individuals was fluctuated from 400 up to 880 pixels (GE manufacturer) and those of bony fat tissue from 280 to 340 pixels. On the contrary, the affected values ranged from 80–130 pixels. In order to reduce the possibility of false positive and false negative values we combined all the region of interest (ROI) measurements with the degree of infiltration detected on T1W images. Then, we combined the above MRI findings and introduced the following classification: stage I: ROI 1/2 of normal values and bone infiltration up to 30%; stage II: ROI 1/3 of normal values and bone infiltration from 30 to 60%; stage III: ROI 1/4 of normal values and bone infiltration from 60% to 80% and stage IV: detection of epiphyseal infiltration, osteonecrosis and deformity regardless of the ROI’s values. Nevertheless in all patients the ROIs fluctuated from 80–130 pixels.Before the initiation of replacement therapy, 12 (46%) patients had bone pain, while 3 patients had restrictions in free movement and 2 patients experienced difficulty in walking at the time of study initiation. The majority of the patients (91%) had elevated chitotriosidase levels (median: 1647 nmol/ml/hr, range: 147–18880 nmol/ml/hr) indicating active disease, while 25% of the patients had abnormally high levels of acid phosphatase. All but one patient had abnormal MRI findings: 3 (11%) had stage I MRI abnormalities, 18 (66%) had stage II, 3 (11%) stage III and 2 (7%) had stage IV abnormalities in MRI. Twelve months post-imiglucerase therapy, all patients had reduction in pain and improvement in physical activity. Regarding MRI data, 7 (26%) of patients remained stable (had the same MRI stage), 14 (52%) showed an improvement in MRI stage (at least one stage down in our staging system), while 6 (22%) patients had a deterioration in MRI findings on femoral bones.Our data suggest that imiglucerase, after one year of therapy, improves symptoms in all patients with bony complications due to non-neuronopathic GD and improves MRI findings in almost half of them. The semi-quantitative MRI method that we used is relatively easy to be performed compared to other available methods for GD and would be useful for the evaluation and follow-up of bone marrow involvement in patients with non-neuronopathic GD. Disclosures:No relevant conflicts of interest to declare.

Bone Involvement in Patients with Non-Neuronopathic Gaucher Disease Is Characterized by Increased Osteoclast Function Partially Due to High Circulating C-C Motif Ligand 3,

Abstract 3215Gaucher disease (GD) is a lysosomal storage disease characterized by a genetic disruption in the metabolic breakdown of glucocerebroside caused by a lack of the enzyme beta-glucocerebrosidase. Bone complications are common in the non-neuronopathic form of GD (formerly type I) and lead to considerable pain, limitations in mobility and negative impact on the quality of life. Skeletal manifestations of GD include osteopenia and osteoporosis, bone loss, Erlenmeyer flask deformity and osteonecrosis. Low bone mineral density (BMD) is thought to arise from enhanced bone resorption mediated through macrophage-derived factors, such as the C-C motif ligand 3 (CCL3) chemokine. The aim of this study was to evaluate the clinical and biochemical characteristics of bone involvement in patients with the non-neuronopathic form of GD.We studied 27 adult patients (15M/12F, median age 44.5 years, range: 18–71 years) with non-neuronopathic GD. Three patients were evaluated at diagnosis, while the others after a 6-month discontinuation of replacement therapy with imiglucerase due to temporary shortage of the drug supply in Greece. BMD of the lumbar spine and femoral neck was determined in all patients, using dual energy X-ray absorptiometry (DXA). Furthermore, magnetic resonance imaging (MRI) of the femoral bones was also performed in all patients. The MRI protocol included T1-weighted spin echo and turbo echo sequences, short time inversion recovery (STIR) sequences and T2-weighted spin echo, turbo spin echo (TSE) sequences. Based on the severity of abnormal MRI findings and the decreased fat content in the bone marrow, patients were categorized in 4 grades: I-IV. On the day of MRI, we evaluated also the circulating levels of CCL3 along with two sensitive serum markers of bone resorption: C-telopeptide of collagen type-1 (CTX) and tartrate resistant acid phosphatase isoform type-B (TRACP-5b), which is produced mainly by activated osteoclasts. Bone markers were also evaluated in 25 healthy, gender- and age- matched controls, as well as in all patients, 6 months post re-initiation of replacement therapy (when the drug became available again).Twelve patients (46%) had bone pain, while 3 patients had restrictions in free movement and 2 patients experienced difficulty in walking at the time of study initiation. The majority of the patients (91%) had elevated chitotriosidase levels (median: 1647 nmol/ml/hr, range: 147–18880 nmol/ml/hr; UNL: 150 nmol/ml/hr) indicating active disease, while 25% of the patients had abnormally high levels of acid phosphatase. Six (22%) patients had osteoporosis at least in one of the studied sites, 18 (66%) patients had osteopenia and 3 patients had normal DXA scan. All but one patient had abnormal MRI findings: 3 (11%) had grade I MRI abnormalities, 18 (66%) had grade II, 3 (11%) grade III and 2 (7%) grade IV. Erlenmeyer flask sign was observed in 23 (85%) patients.At study initiation, GD patients had elevated serum levels of TRACP-5b (mean±SD: 7.6±5.3 IU/L) compared to controls (1.3±0.6 IU/L; p<0.001) and high circulating CCL3 (71±31 pg/ml) versus controls (16±11 pg/ml, p<0.001). There was a positive correlation between serum TRACP-5b and chitotriosidase levels (r=0.493, p=0.017) as well as between TRACP-5b and CCL3 levels (r=0.512, p=0.01). Patients with osteopenia or osteoporosis had elevated chitotriosidase levels (4741±4876 nmol/ml/hr) compared to others (478±415 nmol/ml/hr; p=0.035). Patients with advanced MRI disease (grades III & IV) had elevated levels of acid phosphatase (13.1±3.7 U/L) compared to grades I & II (8.6±1.8 U/L; p=0.011). Six months post- re-initiation of replacement therapy TRACP-5b levels (7.1±4.1 IU/L; p=0.011) and CCL3 levels (42±29 pg/ml; p=0.01) were significantly reduced.In conclusion, our findings suggest that patients with active non-neuronopathic GD have abnormal MRI bone findings in the femoral bones and increased incidence of bone loss (osteopenia or osteoporosis). The enhanced osteoclast activity (assessed by the high TRACP-5b serum levels) which is present in GD, is - at least partially - due to elevated CCL3 (a potent osteoclast activator) and it seems to be implicated in the biology of bone involvement in non-neuronopathic GD. Replacement therapy reduces abnormal osteoclast function; longer follow-up will reveal if this reduction is followed by the reversal of bone involvement in this disease. Disclosures:No relevant conflicts of interest to declare.

Bone pathology inpsoriatic arthritis

Objective. To study different variants of osteolysis in pts with psoriatic arthritis (PA) and to reveal their relationship with other clinico-radiological features of joint damage. Material and methods. 370 pts with definite PA having different variants of joint damage were included. Radiological examination of bones and joints (in some cases large picture frame) was performed. Morphological evaluation of synovial biopsies was done in 34 pts with PA and 10 pts with rheumatoid arthritis (RA). Results. Different types of osteolysis were revealed in 80 (21,6%) pts. Osteolytic variant of joint damage was present in 29 pts. 33 pts had acral, 48 — intra-articular osteolysis and 16 - true bone atrophy. Frequency and intensity of bone resorption were associated with severity of PA. Acral osteolysis correlated with arthritis of distal interphalangeal joints and onychodystrophy. Intra-articular osteolysis was most often present in distal interphalangeal joints of hands and metacarpophalangeal joints (39,6% and 41,7% respectively). Characteristic feature of PA was combination of prominent resorption with formation of bone ankylosis and periosteal reaction. Ankylosis was present in 33,3% of pts with intra-articular osteolysis and in 60% of pts with combination of different osteolysis variants. Systemic reaction of microcirculation in synovial biopsies was most prominent in osteolytic variant: marked thickening of capillary and venule basal membrane with high level of acid phosphatase, increased capillary and precapillary blood flow with stasis features, vascular lymphocyte and macrophage infiltration, productive vasculitis with annular wall thickening, thrombovasculitis and villi deep layer sclerosis. Conclusion. Different variants of osteolysis show bone involvement in PA. Acral and intra- articular osteolysis association with bone ankylosis and periostitis proves their common pathogenetic entity.

Serum acid phosphatase as a tumour marker in giant cell tumour of bone.

The serum acid phosphatase value was examined in nine patients with giant cell tumour of bone. Five showed a high level of acid phosphatase, which fell to within normal limits after surgery. Although the remaining four patients showed a normal acid phosphatase level before surgery, the postoperative acid phosphatase level was lower than the preoperative level in each case. Therefore, it is strongly suggested that serum acid phosphatase is a useful tumour marker in diagnosing giant cell tumour of bone as well as in evaluating the efficacy of treatment.

Hydrolysis of triclosan monophosphate by dental plaque and selected species of oral micro-organisms.

Triclosan monophosphate is a phosphorylated derivative of the antimicrobial agent, triclosan. In comparison with triclosan, it is highly soluble in aqueous solutions. It is hypothesized that, within the oral environment, triclosan monophosphate (which may be devoid of antimicrobial activity) will be hydrolyzed into triclosan by the action of microbial phosphatases. The liberated triclosan may then exert antimicrobial activity. To test this hypothesis, we designed experiments to measure the phosphatase activity of plaque and selected species of oral micro-organisms and to demonstrate hydrolysis of triclosan monophosphate. Tests comparing the minimal inhibitory concentration and minimal bactericidal concentration of triclosan and triclosan monophosphate were also undertaken. Dental plaque and the majority of the bacterial strains tested showed phosphatase activity against p-nitrophenyl phosphate which peaked below neutral pH (acid phosphatases) or above neutral pH (alkaline phosphatases). Dental plaque showed the highest levels of alkaline phosphatase (optimum at pH 9.0) and relatively high levels of acid phosphatase (optimum at pH 6.0 to 6.5). Dental plaque and selected species of micro-organisms were all capable of hydrolyzing triclosan monophosphate, albeit at different rates. The minimal inhibitory concentration and minimal bactericidal concentration values for triclosan monophosphate against eight bacterial strains were always considerably higher than the corresponding values for triclosan. Addition of triclosan monophosphate to an established culture (ca. 10(9) cfu/mL) of Capnocytophaga gingivalis growing continuously showed that triclosan monophosphate was rapidly hydrolyzed into triclosan with concomitant loss of total bacterial viability. It is therefore likely that triclosan monophosphate will be broken down into triclosan within the oral environment with concomitant antimicrobial activity.

Phosphatase production and activity in Citrobacter freundii and a naturally occurring, heavy-metal-accumulating Citrobacter sp.

The ability of a naturally occurring Citrobacter sp. to accumulate cadmium has been attributed to cellular precipitation of CdHPO4, utilizing HPO4(2-) liberated via the activity of an overproduced, Cd-resistant acid-type phosphatase. Phosphatase production and heavy metal accumulation by batch cultures of this strain (N14) and a phosphatase-deficient mutant were compared with two reference strains of Citrobacter freundii. Only strain N14 expressed a high level of acid phosphatase and accumulated lanthanum and uranyl ion enzymically. Acid phosphatase is regulated via carbon-starvation; although the C. freundii strains overexpressed phosphatase activity in carbon-limiting continuous culture, this was approximately 20-fold less than the activity of strain N14 grown similarly. Citrobacter strain N14 was originally isolated from a metal-contaminated soil environment; phosphatase overproduction and metal accumulation were postulated as a detoxification mechanism. However, application of Cd-stress, and enrichment for Cd-resistant C. freundii ('training'), reduced the phosphatase activity of this organism by about 50% as compared to Cd-unstressed cultures. The acid phosphatase of C. freundii and Citrobacter N14 had a similar pattern of resistance to some diagnostic reagents. The enzyme of the latter is similar to the PhoN acid phosphatase of Salmonella typhimurium described by other workers; the results are discussed with respect to the known phosphatases of the enterobacteria.

High-level expression of endogenous acid phosphatase inhibits growth and vectorial secretion in Saccharomyces cerevisiae.

The secretion pathway of Saccharomyces cerevisiae was challenged by constitutively overexpressing plasmid-encoded acid phosphatase, a secreted endogenous glycoprotein. A 2-microns-based multicopy plasmid carrying the coding sequence of acid phosphatase under the control of a truncated variant of the strong constitutive glyceraldehyde-3-phosphate dehydrogenase promoter was used for expression. Selection for the promoterless dLEU2 marker leads to a growth arrest. This is not per se due to leucine starvation, but due to intracellular accumulation of highly glycosylated enzymatically active acid phosphatase. Immunofluorescence and cytological analysis indicate that intracellular accumulation of acid phosphatase occurs in a subpopulation of cells. By Ludox-AM density centrifugation, these cells can be enriched on the basis of their higher density. The dense accumulating cells have a higher average plasmid copy number and produce more acid phosphatase than non-accumulating cells of low density. These cells are defective in directed secretion and bud formation, therefore can no longer grow and show dramatic changes in cell morphology. We suggest that the secretion pathway in these cells is overloaded with the high level of acid phosphatase leading to a shutdown in vectorial secretion, subsequently to a standstill in growth and to the intracellular accumulation of further expressed acid phosphatase. We have indications that accumulation of acid phosphatase occurs in the late Golgi, suggesting a limitation of the overall secretion at this stage.

Acid phosphatase activity of virulent and avirulent clones of Leishmania donovani promastigotes.

Virulent and avirulent clones of Leishmania donovani promastigotes were examined for their acid phosphatase activity. The acid phosphatase activity of whole-cell lysates of virulent clones was 1.5 to 2.0 times higher than that of avirulent clones. Pellet fractions (260,000 x g, 30 min) from sonicated promastigotes of a virulent clone and an avirulent clone contained 60 and 40% of the total enzyme activity, respectively. Membrane-bound acid phosphatase was extracted with Triton X-100 from the pellet. This membrane-bound phosphatase activity was 2.4-fold higher in virulent organisms than in avirulent organisms. The membrane acid phosphatase exhibited two distinct bands on polyacrylamide gels stained for enzyme activity. One diffuse, faster-migrating band showed identical electrophoretic mobility in both virulent and avirulent clones, although a higher enzymatic activity was observed with the extract from virulent cells. In contrast, a slower-migrating band was different between the two clones in the mobility. These results suggest that membrane-bound acid phosphatase was quantitatively and qualitatively different between virulent and avirulent promastigotes of L. donovani. In addition, virulent cells produced a relatively high level of acid phosphatase throughout the growth in culture.

The Examination of Vaginally Inserted Plastic Tampon Applicators for Genetic Markers and Evidence of Prior Sexual Intercourse

Vaginally inserted plastic tampon applicators were obtained from 42 female volunteers. The applicators were examined for the presence of ABH blood group substances, phosphoglucomutase (PGM), amylase, acid phosphatase, P30, and intact spermatozoa. Each applicator was accompanied by a control blood sample, a saliva specimen, a brief sexual and menstrual history, and method of birth control of the donor. Eight of the male sexual partners of the donors submitted blood and saliva samples. One male sexual partner submitted only a saliva sample. ABH blood group substances corresponding to the donor were recovered from 36 of the 42 applicators. The remaining 6 applicators revealed a combination of the donor's and sexual partner's ABH substances. The female's PGM type was recovered from 34 of the applicators. The remaining 8 applicators failed to show PGM activity. Of the applicators, 15 indicated evidence of prior sexual intercourse by the detection of ABH substances not consistent with the applicator donor (6 samples), high levels of acid phosphatase (11 samples), or recovery of spermatozoa (8 samples) or some combination of these. All applicator samples failed to show the presence of either P30 activity or PGM factors foreign to the female.

Serum acid phosphatase elevation associated with transurethral resection syndromes

In 6 patients undergoing transurethral resection of the prostate for benign prostatic hyperplasia symptoms of post-transurethral prostate resection syndrome developed. Serum acid phosphatase determinations in the recovery room showed that all patients had high acid phosphatase levels although each had normal levels preoperatively. All patients showed a normal acid phosphatase level on the first postoperative day. The acid phosphatase elevations indicate significant intraoperative absorption of prostate tissue substances. The association of clinical symptoms with enzyme elevation suggests that the etiology of the confusing clinical syndromes following transurethral prostate surgery may be due to the intravenous absorption of not only irrigant solution but also tissue substances from the prostate gland.

Calcitonin effects on isolated bone cells.

Calcitonin decreased calcium uptake in specific rat bone cell populations obtained by sequential collagenase digestions of calvaria. The calcitonin effect on calcium uptake was observed in the same populations that manifested calcitonin-induced increases in cyclic AMP as well as high levels of acid phosphatase and the ability to release 45Ca from prelabeled devitalized bone. No effect of calcitonin was observed in cell populations that had high levels of alkaline phosphatase and lacked the potential to resorb devitalized bone. These results suggest that changes in cell calcium as well as cyclic AMP may be involved in the mode of action of calcitonin on osteoclast-like cells.

Prostacyclin: Effects on Cyclic AMP in Bone Cells

Prostacyclin (PGI2) increased cyclic AMP in specific rat bone cell populations obtained by sequential collagenase digestion of calvariae. An osteoclastic population, characterized by high acid phosphatase levels and the ability to resorb bone in vitro, was more sensitive as reflected by responses to lower doses of PGI2 (2.3 x 10(-8)M) as well as greater responses to higher doses (3.5 x 10(-4)) of PGI2 than the other populations isolated. The osteoblastic populations, characterized by high alkaline phosphatase levels and the inability to resorb devitalized bone did not respond significantly to PGI2 at doses less than 10(-4)M and increases in cAMP were smaller. Then results suggest a differential effect of PGI2 on osteoclasts and osteoblasts which may be involved in the mode of action of endogenously produced PGI2 in bone.

Pre-fertilization ovule development inCapsella: ultrastructure and ultracytochemical localization of acid phosphatase in the meiocyte

Pre-meiotic and prophase I ovules ofCapsella bursa-pastoris (L.) Medic.(monosporic,Polygonum type of gametophyte development) were fixed routinely or incubated in a modified Gomori medium containing β-glycerophosphate as a substrate. Prior to the beginning of meiosis the potential meiocyte is ultrastructurally similar to the other cells of the nucellus and is distinguished only by its size and position. At the initiation of prophase I dramatic ultrastructural and ultracytochemical changes take place in the female meiocyte. These include the sudden appearance of cytoplasmic structures composed of single and multiple concentric cisternae, distinctive changes in plastids and mitochondria, and the blebbing of 0.3 μm double-membraned vesicles from the nuclear envelope. The concentric cisternae encapsulate portions of cytoplasm containing ribosomes, plastids, mitochondria, ER fragments and vesicles. Both single and multiple concentric cisternae localize high levels of acid phosphatase and function as autophagic vesicles (AVs) that sequester ribosomes and organelles for destruction during meiosis. Plastids stop dividing and become more spherical during prophase I. Some plastids localize acid phosphatase and many show continuities between the outer membrane and the plastid envelope and acid phosphatase-rich RER cisternae. Mitochondria appear as dense, contracted spheres or rods. Some mitochondria localize acid phosphatase but they do not show membrane confluencies with the ER. Some of the plastids and mitochondria that are segregated into the functional megaspore at meiosis II are destroyed but others apparantly survive meiosis and give rise to the plastid and mitochondrial populations of the young gametophyte (Schulz andJensen, unpublished). The lateral and end walls of the meiocyte show patches of intense aniline blue fluorescence and the chalazal end wall of the cell is perforated with large numbers of plasmodesmata.

Investigations of Vacuoles Isolated from Tobacco

Nicotine was shown to be associated with mature vacuoles isolated from protoplasts of Nicotiana rustica. The vacuolar preparations also contained high levels of acid phosphatase, ATPase, and approximately 30% of the soluble protoplastic protein. The contamination of the vacuolar isolate by chlorophyll, succinate dehydrogenase, and NADPH cytochrome c reductase (markers for chloroplasts, mitochondria, and endoplasmic reticulum) was low. The enzymic activity associated with the vacuoles was not due to the exogenously supplied digestive enzymes used in the preparation of the protoplast. The relatively easy isolation of tobacco vacuoles makes this an excellent system for biochemical investigations of the vacuole.

Histochemistry and ultrastructure of adrenal cortical development in the golden hamster

Adrenal glands were collected from fetuses of golden hamsters at 12 1 2 , 14 1 2 , and 15 1 2 days' gestation and from newborn animals. Samples were prepared for light microscopy, histochemical analyses, and electron microscopy. Ultrastructural differentiation of the fetal adrenal was characterized by the development of zonation and typical steroid-producing cells. At 12 1 2 days' gestation, cortical cells had smooth borders, many polyribosomes, and contained mitochrondria with lamellar cristae. Smooth endoplasmic reticulum was not evident. Signs of functional zonation into deep and superficial zones were evident by 14 1 2 days' gestation. Cortical cells of the superficial zone contained scattered lipid droplets, small Golgi areas, and mitochrondria with lamellar cristae. This zone later developed into the adult zona glomerulosa. The cortical cells of the deep zone were characterized by mitochondria containing tubulovesicular cristate, well-developed Golgi areas, vesicular smooth endoplasmic reticulum, and lysosomes. At 15 1 2 days' gestation and in newborn animals the zonation was more prominent, and desmosomes and tight junctions were present between the deep cortical cells. High levels of acid phosphatase, alkaline phosphatase, and esterase were present in the deep cortex, but lower activity occurred in the superficial zone. The activity of 3β-hydroxysteroid dehydrogenase was low in all specimens.