Rapid Identification of Legionella Pathogenicity by Surface-Enhanced Raman Spectroscopy / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To establish Surface-enhanced Raman Spectroscopy (SERS) can be used as a rapid and reliable method to distinguish virulent strain and mild strain of L. pneumophila.</p><p><b>METHODS</b>Mortality data were collected from company departments through administrative documents, death certificates, etc. Trend analyses of cancer mortality were performed on the basis of 925 cancer deaths between 2001 and 2010.</p><p><b>RESULTS</b>Our results indicated that the peaks of high virulence strains reached ⋝4000. This criterion was verified by subsequent cell experiments. In addition, we also conducted SERS rapid identification on the virulence of several collected clinical strains and obtained accurate results.</p><p><b>CONCLUSION</b>The present study indicates that the established SERS protocol can be used as a rapid and reliable method to distinguish virulent and mildly virulent strains of L. pneumophila, which can be further used in clinical samples.</p>

A Case of Cystinuria with Multiple Renal Stones in an 8-month-old Girl

Cystinuria is an autosomal recessive disease characterized by impaired transport of cystine and dibasic amino acids in the proximal renal tubule, resulting in the formation of cystine stones. It is believed to account for about 1% of all kidney stones and up to 10% of pediatric stones. Here we report a case of cystinuria with multiple renal stones confirmed by genetic mutational analysis. An 8-month-old girl was admitted to AMC with persistent fever and multiple renal stones. A renal sonogram showed multiple stones at the right renal pelvis, right distal ureter, and left renal medullary portion. An approximately 1 cm renal stone was extracted spontaneously, and stone analysis revealed it to be composed entirely of cystine. Cystinuria was confirmed by increased urine dibasic amino acid levels, including cysteine, and genetic mutational analysis showed the patient to be a homozygote for the pathogenic c. 1820del (p.L607fs) of SLC3A1. Despite treatment with oral hydration and urinary alkalinization, and restricted intake of animal protein, the stones increased in size and number. The patient has since been treated with tiopronin.

Hepatic Ischemic Preconditioning Provides Protection Against Distant Renal Ischemia and Reperfusion Injury in Mice

We previously demonstrated that there are acute and delayed phases of renal protection against renal ischemia and reperfusion (IR) injury with renal ischemic preconditioning (IPC). This study assessed whether hepatic IPC could also reduce distant renal IR injury through the blood stream-mediated supply of reactive oxygen species (ROS). Male C57BL/6 mice were randomly divided into four groups: group I, sham operated including right nephrectomy; group II (IR), left renal ischemia for 30 min and reperfusion injury; group III (IPC-IR), hepatic ischemia for 10 min followed by 10 min of reperfusion before left renal IR injury; group IV (MPG - IPC + IR), pretreated with 100 mg/kg N-(2-mercaptopropionyl)-glycine (MPG) 15 min before hepatic IPC and left renal IR injury. Renal function, histopathologic findings, proinflammatory cytokines, and cytoprotective proteins were evaluated 15 min or 24 hr after reperfusion. Hepatic IPC attenuated the expression of proinflammatory cytokines, tumor necrosis factor alpha, intercellular adhesion molecule 1, and induced inducible nitric-oxide synthase, and the phosphorylation of Akt in the murine kidney. Renal function was better preserved in mice with hepatic IPC (group III) than groups II or IV. Hepatic IPC protects against distant renal IR injury through the blood stream-delivery of hepatic IPC-induced ROS, by inducing cytoprotective proteins, and by inhibiting inflammatory reactions.

Effects of reactive nitrogen metabolites on NK cell-mediated killing of K562 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effects of the exogenous and endogenous reactive nitrogen metabolites (RNM) as NK cell inhibitors on NK cell-mediated killing of K562 cells and the influence of Tiopronin (TIP), glutamylcysteinylglycine (GSH) and histamine dihydrochloride (DHT) as RNM scavengers on reversing the suppressing effect of RNM.</p><p><b>METHODS</b>The exogenous ONOO(-) was administered in the NK+K562 culture system, then the RNM scavengers were added in the NK+K562+ONOO(-) culture system, respectively. The concentrations of RNM, TNF-beta and IFN-gamma, K562 cell inhibition rate (KIR) and the percentage of living NK cells were examined. IL-2+PHA were used as monocyte (MO) activators in the culture system of MO+NK+K562. Then TIP, GSH and DHT were administered and the parameters of NK cell activity were analyzed.</p><p><b>RESULTS</b>After exogenous ONOO(-) was administered in NK+K562 culture system, the percentage of living NK cells was decreased from (93.17 +/- 2.57)% to (71.87 +/- 1.02)% (P < 0.01) and KIR was decreased from (67.47 +/- 2.64)% to (43.44 +/- 2.87)% (P < 0.01). When TIP, GSH and DHT were administered into the systems, the percentage of living NK cells was increased to (91.13 +/- 3.67)% (P < 0.05), (88.03 +/- 1.46)% (P < 0.05), (73.60 +/- 2.76)% (P > 0.05), respectively; KIR was increased to (61.58 +/- 1.89)% (P < 0.05), (60.68 +/- 2.07)% (P < 0.05) and (45.26 +/- 3.31)% (P > 0.05), respectively. When IL-2/PHA were administered in the NK+K562+MO culture system, RNM products was increased from (82.10 +/- 6.60) micromom/L to (193.65 +/- 5.95) micromom/L(P < 0.01);KIR was decreased from (90.64 +/- 3.06)% to (61.29 +/- 2.22)% (P < 0.01). When the TIP, GSH and DHT were administered in the systems, RNM products were decreased to (91.32 +/- 6.81) micromom/L (P < 0.05), (84.66 +/- 5.99) micromom/L (P < 0.05) and (188.92 +/- 5.00) micromom/L (P > 0.05), respectively; KIR was increased to (84.31 +/- 4.56)%(P < 0.05), (81.65 +/- 3.09)% (P < 0.05) and (72.20 +/- 4.10)% (P < 0.05), respectively.</p><p><b>CONCLUSION</b>NK Cell-mediated killing of K562 cells can be suppressed by exogenous and endogenous RNM administration. Both of TIP and GSH can protect NK cells by scavenging RNM and enhance the antineoplasmic activity of NK cells.</p>

Bioequivalence of tiopronin enteric capsules in healthy volunteers / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the bioequivalence of tiopronin enteric capsules (testing preparation, T) versus tablets (reference preparation, R).</p><p><b>METHODS</b>A single oral dose of tiopronin enteric capsules or tablets at 200 mg was administered in 2 groups of Chinese healthy volunteers (n=9) in a randomized crossover design at the interval of 2 weeks. The plasma concentrations of tiopronin were measured by HPLC-MS/MS, and the pharmacokinetic parameters were calculated by DAS 2.0 program. The bioequivalence between the two preparations was evaluated.</p><p><b>RESULTS</b>The main pharmacokinetic parameters were as follows: C(max)(microg.ml(-1)) 3.612-/+1.2393 (R), 3.644-/+1.540 (T); t(max) 4.333-/+1.0853 (R), 3.611-/+1.420 (T); t((1/2))(h) 18.245-/+11.270 (R), 23.403-/+10.500 (T); AUC0-t (microg.h.ml(-1)) 18.732-/+6.92318 (R), 18.713-/+6.585 (T); AUC0-infinity (microg.h.ml(-1)) 21.900-/+7.31220 (R), 20.780-/+7.965 (T). The relative bioavailability of tiopronin enteric capsule was 103.712-/+23.956%, with 90% confidential intervals of ln(AUC0-->72), ln(AUC0-infinity) and ln(C(max)) of 91.1%-111.8%, 96.8%-118.3%, and 85.1%-113.0%, respectively.</p><p><b>CONCLUSION</b>The tiopronin enteric capsules were bioequivalent to the tablets.</p>

Determination of tiopronin in rat plasma by HPLC following fluorescent derivatization / 药学学报

A sensitive, rapid method for determining reduced tiopronin concentration in rat plasma has been developed by using a high-performance liquid chromatography (HPLC) technique in conjunction with the derivatizing agent N-(1-pyrenyl) maleimide (NPM). The analytes were separated on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) using 0.2% glacial acetic acid aqueous solution including 0.015 mol x L(-1) KH2PO4 and acetonitrile (56:44) as a mobile phase at a flow-rate of 0.8 mL x min(-1), and fluorescence detection wavelength were set at lamda(e x) = 340 nm and lamda(e m) = 375 nm, the column temperature was 30 degrees C. The calibration curve was found to be linear over a range of 0.1 - 10.0 microg x mL(-1), the limit of quantitation was 0. 1 mg x L(-1). The coefficients of the variation for the within-run and between-run precisions ranged from 5.3% to 10.8% and 7.0% to 10.8%, respectively. The percentage of absolute recovery ranged from 73.7% to 79.7%. The method was used to determine the concentration of tiopronin in rat plasma after a single intragastric administration of 25 mg x kg(-1) tiopronin to 6 healthy male Wistar rats. The pharmacokinetic process was fitted to a two-compartment model. The method has been successfully applied to the determination of tiopronin in rat plasma.

Scavenger of reactive oxygen metabolites reverses the ROM induced inhibition of NK cell-mediated killing effect on K562 cell in vitro / 中国实验血液学杂志

To investigate the effect of a new reactive oxygen metabolites (ROM) scavenger as immune adjuvant in NK cell-mediated killing effect on K562 cell, IL-2 and PHA were used to activate monocyte to produce ROM, and different concentrations of tiopronin as ROM scavenger was used in the cultivated systems with different ratio of monocytes plus NK cells and K562 cells, while histamine dihydrochloride (DHT) with different concentrations was used as positive control. The reuslts indicated that after IL-2 and PHA were supplemented in the cultivated systems mixing with NK cells and K562 cells as the E/T ratio was 10/1, the ROM production increased from 33.17 +/- 25.02 U/ml to 223.59 +/- 59.41 U/ml (P < 0.05) while K562 cell inhibition rate (KIR) increased from 65.56% to 85.89% (P < 0.05). When the monocytes as the E/MO ratios of 10/2, 10/5 and 10/10 were supplemented respectively, ROM production increased correspondingly (ROM production was 389.79 +/- 43.83 U/ml, 456.74 +/- 42.77 U/ml, 601.42 +/- 21.92 U/ml, respectively), and KIR was on the other round (KIR was 82.36%, 81.36%, 48.09% respectively). Tiopronin, DHT were used in the K562 + NK + MO + IL-2/PHA cultivated systems as the E/MO ratio was 10/2, the ROM production also decreased from 389.79 +/- 43.83 U/ml to -1.20 +/- 60.70 U/ml, 50.21 +/- 22.4 U/ml (P < 0.05), respectively, however KIR increased from 82.53% to 96.09% and 94.64% either (P < 0.05). Higher concentrations of tiopronin and DHT were used, ROM production decreased accordingly. There showed a reverse correlation between ROM production and KIR (r = -0.518). When E/MO ratio was 10/5 or 10/10, tiopronin at any testing concentration and DHT at the higher testing concentration could reduce the ROM production (P < 0.05), but did not improve KIR significantly (P > 0.05). Tiopronin was as good as DHT in ameliorating KIR (P > 0.05) and better than DHT in scavenging ROM (P < 0.05). It is concluded that (1) Monocytes are the major resources of ROM, and the ROM derived from monocytes can disable NK cells in killing neoplasm cells (K562 cells); (2) A new ROM scavenger, tiopronin, can scavenge ROM effectively, and reverse the ROM induced inhibition of NK cell-mediated killing of K562 cell in a certain extent. And tiopronin is better than DHT in scavenging ROM, and as good as DHT in up-regulating KIR. The new ROM scavenger tiopronin with less side effect may take the place of DHT as adjuvant during the adoptive immuno-therapy in leukemia.

Antioxidant Effects of Sulfur-Containing Amino Acids

Sulfur is an essential element for the entire biological kingdom because of its incorporation into amino acids, proteins and other biomolecules. Sulfur atoms are also important in the iron-containing flavoenzymes. Unlike humans, plants can use inorganic sulfur to synthesize sulfur-containing amino acids. Therefore, plants are an important source of sulfur for humans. Sulfur-containing compounds are found in all body cells and are indispensable for life. Some of sulfur-containing antioxidant compounds are, cysteine, methionine, taurine, glutathione, lipoic acid, mercaptopropionylglycine, N-acetylcysteine, and the three major organosulfur compounds of garlic oil, diallylsulfide, diallyldisulfide and diallyltrisulfide. In a comparison of the structure-function relationship among these sulfur-containing antioxidant compounds, dihydrolipoic acid (the reduced form of LA) is the most effective antioxidant. Dihydrolipoic acid contains two sulfhydryl groups and can undergo further oxidation reaction to form lipoic acid. The antioxidative activities of sulfur-containing compounds follow a general trend, the more highly reduced forms are stronger antioxidants and the number of sulfur atoms determine, at least in part, their modulatory activites on the glutathione related antioxidant enzymes. In this article, the antioxidant effects and the antioxidative activities, of sulfur-containing amino acids, are reviewed. In addition, the general antioxidant effects and the structure-function relationship of some sulfur-containing compounds are also reviewed.

Análise da preservação do miocardio após administração de N-2 meercaptopropionilglicina / Analysis of myocardiocardial preservation after B2-mercaptopropionylglicin administration in an experimental model of ischemy and reperfusion in dogs

As doenças cardiovasculares representam a principal causa de morbi-mortalidade da atualidade, sendo a doença arterial coronariana seu maior expoente...

Effects of sulfhydryl compounds on pancreatic cytoprotection in acute necrotic pancreatitis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To observe sulfhydryl compound variation in the injury of pancreatic cells and the effects of external sulfhydryl compounds on cytoprotection.</p><p><b>METHODS</b>Male Wistar mice were divided randomly into three groups: groups A and B served as animal models (retrograde duct infusion with 5% sodium taurocholate), in group A, 45 animals were treated with normal saline therapy, in group B, 45 animals were treated with Tiopronin therapy; and group C, 15 animals, were designated as normal control. Animals were killed at 2, 4, 6, 12 and 24 h, and pancreatic tissue was analyzed for total sulfhydryl (TSH), nonprotein sulfhydryl (NPSH) and malondialdehyde (MDA). Histopathology, serum amylase (Sam) and C reactive protein (CRP) were assessed as well.</p><p><b>RESULTS</b>Levels of Sam and CRP increased in both group A and group B, with corresponding pathological changes of acute nerotic pancreatitis (ANP). Levels of TSH, NPSH and protein sulfhydryl (PSH) in group A decreased markedly during pancreatitis (P < 0.01), but MDA increased significantly (P < 0.01). The depletion of NPSH in group B was markedly ameliorated at 4 h or 6 h, when Tiopronin was prophylactically administered (P < 0.05), after which the level of MDA showed very little increase when compared to group A (P < 0.01). Histopathological damage was attenuated to a certain extent, in regards to serum amylase and CRP.</p><p><b>CONCLUSIONS</b>All sulfhydryl compounds decreased significantly during ANP; external sulfhydryl compound could protect the pancreatic cells most likely as a type of scavengers of oxygen free radicals, which are critically involved in the pathophysiology of ANP. Sulfhydryl plays an important role in the action of pancreatic cytoprotection.</p>

An observation of the effects of Tiopronin on the oxygen free radicals in severely scalded rats receiving delayed fluid resuscitation / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the harmful effects of oxygen free radicals and the protective roles of Tiopronin in severely scalded rats after delayed fluid resuscitation.</p><p><b>METHODS</b>Rats inflicted with 30% III degree scald on the back were employed as the model. They were divided into delayed resuscitation (D) and Tiopronin treatment (T) groups. The changes in superoxide dismutase (SOD) and malonyldialdehyde (MDA) in plasma and subeschar fluid were determined at 24 to 48 postburn hours (PBHs) by means of electron spin resonance (ESR) technique and other routine methods. And the pathomorphological changes in the heart, liver, kidneys and small intestine, and changes in the blood biochemical indices were simultaneously determined. Normal rats were taken as control group (N).</p><p><b>RESULTS</b>The plasma SOD level was was lower than that in N group, while the MDA content in plasma and subeschar fluid in D group was much higher than that in N group. Changes in all the blood biochemical and internal organ pathomorphology were more obvious in the D group. on the other hand, the rat plasma SOD level in T group increased obviously (P < 0.01) while the MDA contents was decreased in T group (P < 0.05) when compared with those rats of D group. In addition, the internal organ pathomorphology and blood biochemical indices were improved evidently in T group.</p><p><b>CONCLUSION</b>Oxidative stress injury was evoked in severely scalded rats after delayed fluid resuscitation, and it could be protected to some extent by Tiopronin.</p>

Lead induced spermatoxicity in mouse and MPG treatment.

Protective efficacy of MPG (2-mercaptopropionyl glycine) was studied against the toxic effects of lead acetate in Swiss albino mice. The animals were treated with single dose of lead acetate @ 180, 200 and 250 mg/kg b.wt. in presence and absence of MPG. The results indicated that the body weight was slightly higher in MPG treated groups on day 10 as compared to only respective lead treated groups in all the three dose level. However, significantly lower body weight was observed in both lead treated and lead along with MPG treated groups as compared to control. Patten of mortality is similar in both lead treated and lead plus MPG treated groups. Conspicuous degenerative changes in testicular tissues and elevation in sperm head shape abnormality were observed in both lead treated and lead along with MPG treated groups but the sperm head shape abnormality and damage were more in lead treated groups as compared to lead plus MPG treated groups. But this difference was non-significant between the two groups. These observations suggest that MPG may not be significantly effective against lead induced damage in testicular tissues at cellular level. However, MPG is able to maintain slightly lower level of sperm abnormality in all the three dose level as compared to their respective lead treated groups. Further, studies are needed to find out the optimum dose of MPG for protection against the lower doses of lead induced lethality as MPG is not significantly effective against the higher doses of lead.

Efeito do antioxidante N2-mercaptopropionilglicina (N2-MPG) na pancreatite aguda experimental / N2-Mercaptopropionylglycine (N2 MPG) in experimental acute pancreatitis

O N2-Mercaptopropionilglicina (MPG) e um potente antioxidante na inibicao da producao de xantina-oxidase. O objetivo deste trabalho foi analisar se este efeito antioxidante poderia propiciar menor agressao do tecido pancreatico na Pancreatite Aguda (PA) induzida por dois processos: dose supramaxima de ceruleina e injecao de taurocolato de sodio a 2,5 por cento no ducto biliopancreatico do rato. Trinta e seis ratos machos Wistar (220-270 g) foram divididos em 2 grupos (G): GI-animais previamente tratados com antioxidante (MPG: 100 mg/kg) 10 minutos antes da inducao da PA e GII-animais sem tratamento previo. Os animais destes dois grupos foram submetidos a PA com dose supramaxima de ceruleina (2 doses de 20ug/kg) e PA com taurocolato onde vinte e seis ratos foram divididos em dois grupos (G): GIII-ratos tratados com MPG 10 minutos antes da PA e GIV-animais sem tratamento previo...

Analise do efeito protetor do N2-mercaptopropionilglicina durante a isquemia-reperfusao de lobo hepatico: estudo experimental em caes e ratos / Protective effect analysis of N-mercaptopropionylglycine during of reperfusion ischemia of hepatic lobe: experimental study in dogs and rats

Um potente inibidor da sintese de radicais superoxidos, a N2-Mercaptopropionilglicina (N2-MPG), principalmente radicais hidroxila (OH), foi testado como agente preventivo na degradacao metabolica e estrutural do parenquima hepatico no processo de isquemia/reperfusao testando a hipotese de participacao significativa da superoxidacao na necrose do figado. Para tanto foram utilizados 22 ratos e 22 caes, distribuidos em dois grupos. Grupo I com administracao de solucao salina 0,9 por cento e Grupo II (GII) com administracao de N2-MPG. As amostras foram submetidas a estudo laboratorial, radiologico, anatomopatologico e estatistico. Os resultados revelaram uma elevacao das transaminases significativamente menor nos animais tratados com N2-MPG...

Cystinuria 3 Cases

We report three cases of cystinuria, presenting with urinary stones. A 2-year-old girl presented with urinary difficulty, hematuria, dysuria of sudden onset, and her 7-month-old younger brother also was presented with urinary difficulty, irritability on urination & stone passage. Other 6-month-old boy was admitted due to sudden onset anuria. They had radioopague renal & ureter stones and stone analysis revealed mixed cystine stones. The diagnosis of cystinuria was confirmed metabolic studies and stone analysis. Lrinary amino acid analysis showed excessive excretion of dibasic amino acids(cystine, ornithine, lysine, arginine). And they all had hypercalciuria and hyperuricosuria. They were treated with combination of percutaneous lithotripsy for large obstructing senes a nd an oral drug therapy with sodium bicarbonate for rendering the urine more alkaline, and alpha-mercaptopropionylglycine(ThiolaR). This form of treatment was sucessful in our three cases with elimination of recurrent nephrolithiasis, but in one patient, nephrotic syndrome possibly caused by ThiolaR was developed. The nephrotic syndrome was recovered spontaneously after cessation of Thiola. A review of literatures was also attempted briefly.

Effect of thiola on methotrexate-induced ultrastructural changes in the rat liver

The effect of the sulfhdryl compound thiola on ultrastructural alterations induced by the anticancer drug methotrexate in the liver of rats was studied. Animals treated with methotrexate at a dose level of 0.004 mg/kg b.w. daily for 1,2 and 5 days showed numerous changes in the hepatocytes. The nuclei showed chromatin and in some cases it was fragmented. Mitochondria were elongated or bizare in shape and the RER were reduced and fragmented. Lysosomes were abundant and glycogen rosettes were depleted. The bile canaliculi appeared dilated with fragmented microvilli. Kupffer cells were hypertrophied indicating hyperactivity. On the other hand, treating animals with methotrexate [at the same dose and for the same periods] followed by injection of thiola [at a dose of 50 mg/kg b.w. daily for one, two and five days], caused an improvement in the fine structure of the hepatocytes. These findings suggest that thiola overcomes the hepatotoxic action of methotrexate

Modulation of glutathione depletion and lipid peroxidation by WR-77913 an 2-mercaptopropionylglycine in cyclophosphamide chemotherapy.

Cyclophosphamide induced a depletion in liver glutathione (GSH) and high rate of lipid peroxidation. GSH depletion was evident from 20 min onwards and the maximum depletion was observed at 3 hr post treatment. Lipid peroxidation was significant only after the maximum depletion of GSH. Pretreatment with either MPG or WR-77913 individually, or in combination could prevent the depletion of GSH and induction of lipid peroxidation after cyclophosphamide treatment.

O uso de MPG na protecao da lesao pos-reperfusao do retalho epigastrico em ilha: estudo experimental em rato / The use of N2 mercaptopropionylglycine in the protection of epigastric skin flaps post reperfusion injury: experimental study in rats

Os autores usaram um removedor de radicais livres, a droga N2-Mercaptopropionilglicina (MPG), na protecao de retalhos epigastricos em ilha de ratos submetidos a um periodo de isquemia seguido de reperfusao. Em 60 ratos Wistar, pesando entre 310 e 380g, foi dissecado um retalho epigastrico de 3,5 x 6 cm a direita do animal. Usando clamps microvasculares obtiveram oclusao da veia epigastrica por cinco horas, a arteria epigastrica por 21 horas ou da arteria e da veia epigastricas por oito horas. Os animais receberam solucao de MPG na concentracao de 100 mg/ml, ou equivalente volume de solucao fisiologica, por via endovenosa, divididas em duas doses iguais, sendo a primeira injetada previamente a elevacao do retalho e a segunda dez minutos antes da retirada dos clamps e consequente reperfusao do retalho. as areas de viabilidade e necrose resultantes foram lidas no setimo dia apos reperfusao do retalho. Os autores concluiram que a droga mostrou ter um efeito protetor dos tecidos dos danos por reperfusao nos tres tipos de oclusao vascular, aumentando significativamente a taxa de sobrevida dos retalhos, quando comparados com os retalhos cujos animais receberam apenas a solucao de cloreto de sodio.

Depletion of ovarian oocytes in sublethally exposed mice to gamma radiation and its modification by 2-mercaptopropionylglycine (MPG).

Adult female Swiss albino mice were exposed to 0.6, 1.2 and 2.4 Gy of 60Co gamma radiations in presence and absence of MPG. Quantitative studies were done in serial sections of ovary at 1, 3, 5, 7, 14 and 35 days after exposure. Primary follicles were found to be most radiosensitive. The depletion in the various types of follicles was checked to some extent by prior administration of MPG, but MPG could not prevent the complete elimination of all types of follicles by the last autopsy interval studied.