Resolution of eleven reported and five novel Podaxis species based on ITS phylogeny, phylogenomics, morphology, ecology, and geographic distribution.

The genus Podaxis was first described from India by Linnaeus in 1771, but several revisions of the genus have left the taxonomy unclear. Forty-four Podaxis species names and nine intraspecific varieties are currently accepted, but most fungarium specimens are labelled Podaxis pistillaris. Recent molecular analyses based on barcoding genes suggest that the genus comprises several species, but their status is largely unresolved. Here we obtained basidiospores and photographs from 166 fungarium specimens from around the world and generated a phylogeny based on rDNA internal transcribed spacer ITS1,5.8S and ITS2 (ITS), and a phylogenomic analysis of 3 839 BUSCO genes from low-coverage genomes for a subset of the specimens. Combining phylogenetics, phylogenomics, morphology, ecology, and geographical distribution, spanning 250 years of collections, we propose that the genus includes at least 16 unambiguous species. Based on 10 type specimens (holotype, paratype, and syntype), four recorded species were confirmed, P. carcinomalis, P. deflersii, P. emerici, and P. farlowii. Comparing phylogenetic analysis with described species, including morphology, ecology, and distribution, we resurrected P. termitophilus and designated neotypes, epitypes, or lectotypes for five previously described species, P. aegyptiacus, P. africana, P. beringamensis, P. calyptratus, and P. perraldieri. Lastly, based on phylogenies and morphology of type material, we synonymized three reported species, P. algericus, P. arabicus, and P. rugospora with P. pistillaris, and described five new species that we named P. desolatus, P. inyoensis, P. mareebaensis, P. namaquensis, and P. namibensis. Citation: Li GS, Leal-Dutra CA, Cuesta-Maté A, et al. 2023. Resolution of eleven reported and five novel Podaxis species based on ITS phylogeny, phylogenomics, morphology, ecology, and geographic distribution. Persoonia 51: 257-279. doi: 10.3767/persoonia.2023.51.07.

[Role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice].

Objective: To investigate the role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice. Methods: The experimental research methods were applied. Eighty-six C57BL/6J male mice (hereinafter briefly referred to as wild-type mice) aged 8-12 weeks were selected for the following experiments. Vγ4 T cells were isolated from axillary lymph nodes of five wild-type mice for the following experiments. Intraperitoneal injection of rapamycin for 42 mice was performed to establish rapamycin-treated mice model for the following experiments. Eighteen wild-type mice were divided into normal control group without any treatment, trauma only group, and trauma+CC chemokine ligand 20 (CCL20) inhibitor group according to the random number table (the same grouping method below), with 6 mice in each group. The full-thickness skin defect wound was made on the back of mice in the latter two groups (the same wound model below), and mice in trauma+CCL20 inhibitor group were continuously injected subcutaneously with CCL20 inhibitor at the wound edge for 3 days after injury. Another 6 rapamycin-treated mice were used to establish wound model as rapamycin+trauma group. On post injury day (PID) 3, the epidermal cells of the skin tissue around the wound of each trauma mice were extracted by enzyme digestion, and the percentage of Vγ4 T cells in the epidermal cells was detected by flow cytometry. In normal control group, the epidermal cells of the normal skin tissue in the back of mice were taken at the appropriate time point for detection as above. Five wild-type mice were used to establish wound models. On PID 3, the epidermal cells were extracted from the skin tissue around the wound. The cell populations were divided into Vγ4 T cells, Vγ3 T cells, and γδ negative cells by fluorescence-activated cell sorter, which were set as Vγ4 T cell group, Vγ3 T cell group, and γδ negative cell group (with cells in each group being mixed with B16 mouse melanoma cells), respectively. B16 mouse melanoma cells were used as melanoma cell control group. The expression of interleukin-22 (IL-22) mRNA in cells of each group was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with the number of samples being 6. Thirty rapamycin-treated mice were used to establish wound models, which were divided into Vγ4 T cell only group and Vγ4 T cell+IL-22 inhibitor group performed with corresponding injections and rapamycin control group injected with phosphate buffer solution (PBS) immediately after injury, with 10 mice in each group. Another 10 wild-type mice were taken to establish wound models and injected with PBS as wild-type control group. Mice in each group were injected continuously for 6 days. The percentage of wound area of mice in the four groups was calculated on PID 1, 2, 3, 4, 5, and 6 after injection on the same day. Six wild-type mice and 6 rapamycin-treated mice were taken respectively to establish wound models as wild-type group and rapamycin group. On PID 3, the mRNA and protein expressions of IL-22 and CCL20 in the peri-wound epidermis tissue of mice in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The Vγ4 T cells were divided into normal control group without any treatment and rapamycin-treated rapamycin group. After being cultured for 24 hours, the mRNA and protein expressions of IL-22 of cells in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively, with the number of samples being 6. Data were statistically analyzed with independent sample t test, analysis of variance for repeated measurement, one-way analysis of variance, Bonferroni method, Kruskal-Wallis H test, and Wilcoxon rank sum test. Results: The percentage of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in trauma only group on PID 3 was 0.66% (0.52%, 0.81%), which was significantly higher than 0.09% (0.04%, 0.14%) in the epidermal cells of the normal skin tissue of mice in normal control group (Z=4.31, P<0.01). The percentages of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in rapamycin+trauma group and trauma+CCL20 inhibitor group on PID 3 were 0.25% (0.16%, 0.37%) and 0.24% (0.17%, 0.35%), respectively, which were significantly lower than that in trauma only group (with Z values of 2.27 and 2.25, respectively, P<0.05). The mRNA expression level of IL-22 of cells in Vγ4 T cell group was significantly higher than that in Vγ3 T cell group, γδ negative cell group, and melanoma cell control group (with Z values of 2.96, 2.45, and 3.41, respectively, P<0.05 or P<0.01). Compared with that in wild-type control group, the percentage of wound area of mice in rapamycin control group increased significantly on PID 1-6 (P<0.01), the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 1 and PID 3-6 (P<0.05 or P<0.01). Compared with that in rapamycin control group, the percentage of wound area of mice in Vγ4 T cell only group decreased significantly on PID 1-6 (P<0.05 or P<0.01). Compared with that in Vγ4 T cell only group, the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 3-6 (P<0.05 or P<0.01). On PID 3, compared with those in wild-type group, the expression levels of IL-22 protein and mRNA (with t values of -7.82 and -5.04, respectively, P<0.01) and CCL20 protein and mRNA (with t values of -7.12 and -5.73, respectively, P<0.01) were decreased significantly in the peri-wound epidermis tissue of mice in rapamycin group. After being cultured for 24 hours, the expression levels of IL-22 protein and mRNA in Vγ4 T cells in rapamycin group were significantly lower than those in normal control group (with t values of -7.75 and -6.04, respectively, P<0.01). Conclusions: In mice with full-thickness skin defects, rapamycin may impair the CCL20 chemotactic system by inhibiting the expression of CCL20, leading to a decrease in the recruitment of Vγ4 T cells to the epidermis, and at the same time inhibit the secretion of IL-22 by Vγ4 T cells, thereby slowing the wound healing rate.

[Emphasis on the diagnosis of chronic kidney disease in diabetes mellitus].

Chronic kidney disease (CKD) in diabetes mellitus includes diabetic kidney disease (DKD), non-diabetic kidney disease (NDKD) or a combination of NDKD and DKD. The clinical and renal pathological manifestations of DKD in type 1 diabetes are different from those in type 2 diabetes. Renal biopsy histopathology is the gold standard for distinguishing DKD from NDKD. However, based on the same pathological diagnosis, DKD patients may still have different disease progression and prognosis due to individual differences in molecular biological mechanisms. Metabonomics, proteomics, transcriptomics and artificial intelligence offer hope for biomarkers to diagnose and predict the progress of DKD.

Circ-0003998 promotes cell proliferative ability and invasiveness by binding to miR-197-3p in osteosarcoma.

OBJECTIVE: The aim of this study was to investigate the level of circ-0003998 in osteosarcoma tissues and cell lines, and to analyze its relation with prognosis of patients, as well as its effect on biological behaviors of osteosarcoma cells. In addition, the potential mechanism of circ-0003998 in promoting osteosarcoma cell proliferation and invasion was explored. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to examine circ-0003998 expression in 60 clinical osteosarcoma tissues and cell lines. The association between circ-0003998 expression and the overall survival rate of patients was explored. After shRNA-circ-0003998 was constructed to down-regulate circ-0003998 expression in osteosarcoma cell lines, the proliferation of osteosarcoma cells was observed through the Cell Counting Kit-8 (CCK-8) and colony formation assay. Meanwhile, cell invasiveness was detected by the transwell invasion assay. Bioinformatics was used to search for microRNAs (miRNAs) that contained the direct effect on circ-0003998. Subsequently, the luciferase reporter vector of circ-0003998 or Krüppel-like factor 10 (KLF10) containing miR-197-3p binding site was constructed. Then, the binding of circ-0003998 or KLF10 to miR-197 was detected using the Dual-Luciferase assay. Furthermore, the function recovery experiment was designed to validate the biological function of circ-0003998 and miR-197 in osteosarcoma. RESULTS: Compared to normal control tissues and cells, the expression of circ-0003998 was significantly up-regulated in both osteosarcoma tissue samples and cell lines. Highly-expressed circ-0003998 was significantly associated with poor overall survival of patients with osteosarcoma. In vitro experiments revealed that the down-regulation of circ-0003998 significantly inhibited the proliferative ability and invasiveness of osteosarcoma cells. Bioinformatics analysis and Dual-Luciferase reporter gene assay indicated that circ-0003998 might bind to miR-197-3p in MG-63, and Saos-2 cell lines. Meanwhile, the functional recovery experiment demonstrated that inhibiting miR-197-3p expression could partially restore the changes in cellular biological behaviors induced by circ-0003998 down-regulation in MG-63 and Saos-2 cells. In addition, miR-197-3p was remarkably down-regulated in osteosarcoma tissues, while KLF10 was up-regulated. However, KLF10 was significantly up-regulated after the knockdown of miR-197-3p in osteosarcoma cells. CONCLUSIONS: Circ-0003998 plays a vital role in promoting the development of osteosarcoma, whose high expression can predict poor clinical prognosis. Circ-0003998 is highly expressed in osteosarcoma tissues and cell lines. The down-regulation of its level can significantly inhibit the proliferative ability and invasiveness of osteosarcoma cells. Meanwhile, circ-0003998 up-regulates the expression of KLF10 by binding to miR-197-3p, thereby promoting osteosarcoma cell growth and invasion, and accelerating the progression of osteosarcoma.

Long non-coding RNA HULC promotes proliferation and osteogenic differentiation of bone mesenchymal stem cells via down-regulation of miR-195.

OBJECTIVE: LncRNAs HULC has been reported to be important regulators in the development of various human diseases. However, the role of HULC in bone mesenchymal stem cells (BMSCs) remains unclear. The present study aimed to explore the regulatory effect of HULC on proliferation and osteogenic differentiation of BMSCs and the underlying mechanism. MATERIALS AND METHODS: The expression of HULC and miR-195 in BMSCs were altered by transfection and measured by qRT-PCR. Cell viability was measured by the CCK-8 assay. Osteogenic differentiation of BMSCs was determined by evaluation of osteogenic markers (Ocn, ALP, Runx2, and Col-1) expression levels using Western blot and qRT-PCR. Furthermore, Western blot was performed to assess the expression of proliferation-related factors, Wnt/ß-catenin and p38MAPK pathway-related factors. RESULTS: HULC overexpression significantly increased cell viability, down-regulated p21 expression but up-regulated CyclinD1 expression, and promoted the levels of osteogenic markers. However, the complete opposite effect was observed in HULC knockdown. Notably, miR-195 expression was negatively regulated by HULC and miR-195 exerted a reversed effect of HULC on BMSCs. Moreover, miR-195 mediated the regulatory effect of HULC on BMSCs proliferation and osteogenic differentiation, as miR-195 mimic abolished the effect of HULC overexpression on BMSCs. We also found that HULC overexpression enhanced the activation of Wnt/ß-catenin and p38MAPK pathway through down-regulating miR-195. CONCLUSIONS: We revealed that HULC promoted proliferation and osteogenic differentiation of BMSCs. The potential mechanism might be involved in its negative regulation on miR-195 and enhanced activation of Wnt/ß-catenin and p38MAPK pathway.

Long non-coding RNA SNHG15 indicates poor prognosis of non-small cell lung cancer and promotes cell proliferation and invasion.

OBJECTIVE: To investigate the expression of human long non-coding ribonucleic acid (RNA) small nucleolar RNA host gene 15 (SNHG15) in non-small cell lung cancer (NSCLC) tissues and its prognostic significance, and to study the influencing mechanism of SNHG15 on biological functions in lung cancer cell lines. PATIENTS AND METHODS: The expression levels of SNHG15 in 49 pairs of lung cancer tissues and para-carcinoma tissues were detected via quantitative real-time polymerase chain reaction (qRT-PCR). The lung cancer cells were transiently transfected with small-interfering (si)-SNHG15 using RNA interference technique. The effect of si-SNHG15 on the proliferation of lung cancer cells was observed via methyl thiazolyl tetrazolium (MTT) assay, its effect on apoptosis of A549 cells was detected via Hoechst 33342 staining and flow cytometry, and its effects on invasion and migration of A549 cells were studied via wound healing assay and transwell assay. RESULTS: Results of qRT-PCR showed that the expression of SNHG15 in cancer tissues was increased compared with that in para-carcinoma tissues. Results of cell counting kit-8 (CCK-8) assay showed that knocking down SNHG15 could significantly inhibit the proliferation of lung cancer A549 cells. Hoechst 33342 staining and flow cytometry revealed that knocking down SNHG15 could significantly promote apoptosis of A549 cells. Wound healing assay and transwell assay revealed that knocking down SNHG15 could significantly inhibit the invasion and metastasis capacities of lung cancer A549 cells. Results of Western blotting showed that knocking down SNHG15 could inhibit the invasion and metastasis of A549 cells through inhibiting the expressions of epithelial-mesenchymal transition (EMT), matrix metalloproteinase-2 (MMP-2) and MMP-9 in cells. CONCLUSIONS: The expression of SNHG15 in lung cancer tissues is significantly higher than that in para-carcinoma tissues, the prognosis of patients accompanied with a high expression of SNHG15 is poor, and knockdown of SNHG15 in A549 cells can inhibit cell proliferation, invasion, and metastasis, and promote apoptosis.

Erythropoietin Receptor Activation Protects the Kidney From Ischemia/Reperfusion-Induced Apoptosis by Activating ERK/p53 Signal Pathway.

BACKGROUND: Apoptosis plays an important role in renal ischemia/reperfusion (IR) injury. Evidence has shown that erythropoietin (EPO) has an antiapoptotic effect. Therefore, this study aimed to explore the effect and potential mechanism of EPO in renal IR injury. METHODS: Kidney IR injury in rats was established by clamping the left renal artery for 30 minutes followed by 24 hours of reperfusion, along with contralateral nephrectomy. Renal function, renal histology, and expression of EPOR, p-EPOR, ERK, p-ERK, p-p53, p53, Bcl-2, Bcl-xl, Bad, and Bax were examined. RESULTS: Pretreatment with EPO significantly reduced renal dysfunction, pathologic change, and expression of Bad and Bax. Furthermore, EPO treatment enhanced the expression of p-ERK, p-p53, Bcl-2, and Bcl-xl with no influence on the expression of EPOR, ERK, and p53. CONCLUSIONS: These findings demonstrated that EPO pretreatment can attenuate renal IR injury by inhibiting apoptosis by promoting activation of the ERK/p53 signaling.

Sodium valproate inhibits MDA-MB-231 breast cancer cell migration by upregulating NM23H1 expression.

Breast cancer is a common cancer in women, with a highly variable course, from inoffensive to lethal. To find a more effective strategy for its treatment, sodium valproate has been tested as an anti-cancer drug; it is the only clinically available histone deacetylase inhibitor. However, data about the effects of sodium valproate on breast cancer are insufficient in both animals and humans; studies have yielded conflicting conclusions. In particular, little is known about the association between expression of the metastasis suppressor Nm23H1 gene and breast cancer. We hypothesized that sodium valproate regulates NM23H1 expression, and affects migration and/or invasion. We found that sodium valproate at concentrations of 0.8-3.2 mM inhibits migration and modulates Nm23H1 gene expression in a concentration-dependent manner. Confluent MDA-MB-231 cells were scratched by a micropipette tip after VPA treatment for 24 h; 24 h later, the scratch was almostly closed in the 0 mM VPA-treated cells, while the 3.2 mM VPA-treated cells migrated the slowest. The cell migration ratio exposed to 0.8, 1.6 and 3.2 mM VPA was about 66.67, 30.67 and 26.67% (P < 0.05). We also found evidence that sodium valproate upregulates NM23H1 expression, which is a clue to its anti-cancer mode of action. The NM23H1 gene expression was relative fold increased determined by Western blotting at 3.2 mM VPA. Collectively, these observations indicate that sodium valproate has potential for use in breast cancer treatment.

Influence of proline-rich inositol polyphosphate 5-phosphatase, on early development of fertilized mouse eggs, via inhibition of phosphorylation of Akt.

OBJECTIVES: Proline-rich inositol polyphosphate 5-phosphatase (PIPP) is one of the signal-modifying enzymes that play pivotal regulatory roles in PI3K signalling pathway. The aim of this study was to determine the role of PIPP in early development of fertilized mouse eggs, via inhibition of Akt activity and subsequent downstream signalling events. MATERIALS AND METHODS: The mRNA transcript levels of endogenous PIPP and Akt1, Akt2, Akt3 were detected in G(1) , S, G(2) and M phases of fertilized mouse eggs by RT-PCR. Levels of exogenous PIPP, phosphorylated Akt at Ser473, dephosphorylated cdc2 at Tyr15 and levels of CCNB1, were detected respectively by immunoblotting. Changes in Akt localization were observed by fluoroimmunoassay; meanwhile, changes in activity of Akt and its downstream MPF were detected. Percentages of cells undergoing division were determined by counting, using a dissecting microscope. RESULTS: PIPP and Akt1 transcripts were detectable in G(1), S, G(2) and M phases of fertilized mouse eggs, but Akt2 and Akt3 were not. We also observed that overexpression of PIPP in fertilized eggs decreased expression of phosphorylated Akt at Ser473 and altered membrane localization of phosphorylated Akt at Ser473 specifically. Furthermore, overexpression of PIPP resulted in decreases in mitosis-phase promoting factor activity, level of dephosphorylated cdc2 at Tyr15 and cleavage rate of fertilized mouse eggs. CONCLUSIONS: Our data suggest, for the first time, that PIPP may affect development of fertilized mouse eggs by inhibition of level of phosphorylated Akt at Ser473 and subsequent inhibition of downstream signal cascades.

Implantation of BM cells transfected with phVEGF165 enhances functional improvement of the infarcted heart.

BACKGROUND: Experimental data have suggested that BM-cell implantation can improve infarcted cardiac function. However, the number of implanted cells that survive is still unknown. The present study was performed to investigate whether implantation of autologous BM mononuclear cells (BM-MNCs) transfected with phVEGF165 can increase the number of surviving implanted cells, and enhance functional improvement of infarcted hearts in rabbits. METHODS: Acute myocardial infarction (AMI) in rabbits was replicated by ligating the left anterior descending coronary artery, and animals were randomly divided into the following three groups: I AMI control group (n=7); II BM-MNCs transfected with phVEGF165 implantation group (n=7); III BM-MNCs implantation group (n=7). In addition, sham-operated (n=5) rabbits were randomly selected to serve as a non-infarction control group (VI). Animals for cell implantation received intramyocardial injections of autologous BM-MNCs 14 days after AMI. Echocardiography and hemodynamic studies were performed to evaluate cardiac structure and function 28 days after implantation. The implanted sites were examined using immunofluorescence to identify the phenotypes and number of the labelled cells. Reverse transcriptase (RT)-PCR and Western blot analysis were performed to detect the expression of hVEGF165 gene and VEGF165 protein respectively. RESULTS: Failed cardiac function produced by AMI was significantly improved in Groups II and III 28 days after cell implantation. BM-MNCs transfected with phVEGF165 implantation conferred a further improvement of cardiac function, with significant changes of all assessed parameters when compared with BM-MNCs implantation alone (all P<0.05). The implanted cells demonstrated myogenic differentiation with the expression of Troponin T and organized contractile proteins. The positive staining for Factor VIII-related Ag indicated the induction of angiogenesis in the infarct area. The percentage of Brdu-positive myocyte, endothelial cells was 75+/-%, 34.1+/-4.6% in Group II, significantly higher than that in Group III(51+/-7%, 11.3+/-2.5% respectively, P<0.05). RT-PCR analysis demonstrated exclusive expression of hVEGF165 gene in Group II, and Western blot showed the expression of VEGF165 protein was significantly higher in Group II than in Group III (P<0.05). CONCLUSIONS: Implantation of BM-MNCs transfected with phVEGF165 can increase the number of implanted cells surviving, and enhance the impaired cardiac function after AMI.

[Effect of allitridi on expression of proliferating cell nuclear antigen of cultured rabbit aortic smooth muscle cell].

OBJECTIVE: To observe the effect of allitridi on the expression of proliferating cell nuclear antigen (PCNA) of cultured rabbit aortic smooth muscle cell (SMC). METHODS: The expression of PCNA was examined with immunohistochemical technique (LSAB method) and the contents of superoxide dismutase (SOD), lipid peroxide (LPO), prostacyclin (PGI2) and cyclic adenosine monophosphate (cAMP) in medium were simultaneously determined. RESULTS: Allitridi has the effects of increasing SOD activity, decreasing LPO, elevating PGI2 and cAMP, reducing the expression of PCNA (all P < 0.05-0.01). CONCLUSION: Allitridi could inhibit SMC proliferation.

[An improved technique for bloodless hepatic resection on the in situ cold perfused liver].

An improved technique for bloodless hepatic resection using in vivo isolation and asanguinous hypothermic perfusion was described to deal with the huge liver tumour involving the liver hilum, the main hepatic veins and retrohepatic inferior vena cava. The original Fortner's technique was modified, including the incision; isolated cold perfusion of the healthy half of the liver through the portal vein; suprahepatic outlet for the perfusate, and shortened hepatic ischemia time by perfusing hepatic artery prior to the repair or reconstruction of the portal vein. Using this technique, two huge liver tumors were successfully resected, and the indications, intraoperative monitoring and early postoperative courses were discussed.

An improved technique for bloodless hepatic resection on in situ cold perfused liver.

An improved technique for bloodless hepatic resection using in situ isolation and asanguinous hypothermic perfusion was described to deal with huge liver tumors involved in the liver hilum, the main hepatic veins and retrohepatic inferior vena cava. The original Fortner's technique was modified, including the choice of incision; semi-isolated perfusion of the liver portion preserved through the single portal vein; suprahepatic outlet of the perfusate and the shortening of the period of hepatic ischemia by reperfusion of hepatic artery prior to the repair or reconstruction of the portal vein. The initial successful experience of the technique applied to 2 pediatric cases with giant liver tumors was reported, and the indications, intraoperative and early postoperative courses were discussed.

Effect of hydrogen peroxide treatment on the nutritional quality of rapeseed flour fed to weanling rats.

The effect of treating rapeseed flours with hydrogen peroxide on the glucosinolate content and nutritional value of the protein was examined. Four flours were prepared from Target variety rapeseed (Brassica napus) by dehulling and defatting the seed (sample RF), by heat treating and water washing the dehulled seed prior to defatting (sample WWRF), and by treating a part of samples RF and WWRF with solutions of 7 and 3% hydrogen peroxide, respectively. Chemical analysis showed that the hydrogen peroxide treatment lowered the glucosinolate content of the flour but was not as effective as the water extraction. The hydrogen peroxide treatment also oxidized methionine to its sulfoxide and sulfone and cysteine to cysteic acid. In the first experiment, weanling rats were fed for 3 weeks diets in which casein or each of the flour preparations provided 5, 10, or 20% protein. Rats fed the high glucosinolate-containing flour (sample RF) at the 10 to 20% protein level died, while those fed 5% survived but lost weight. Those fed the hydrogen peroxide-treated flours survived, but weight gains and food consumption were low compared with the values of the caseinor WWRF-fed groups. Rats fed sample RF exhibited enlarged thyroids. Those fed the peroxide-treated samples had high plasma levels of methionine sulfoxide and sulfone. In the second experiment, additions of 0.15 or 0.30% methionine to the 10% protein diets resulted in increased weight gains of the groups fed the peroxide-treated flours. It was concluded that the hydrogen peroxide treatment was effective in reducing the glucosinolate content of the rapeseed flour. However, the production of the oxidized sulfur amino acids, in particular methionine sulfone, reduced considerably the nutritional value of the protein.