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Biomedical subjects

Ying Yu

Publications and source records attributed to Ying Yu.

At least 55 records · Page 3Linked to original sources

Enhanced expression of transient receptor potential channels in idiopathic pulmonary arterial hypertension.

Pulmonary vascular medial hypertrophy caused by excessive pulmonary artery smooth muscle cell (PASMC) proliferation is a major cause for the elevated pulmonary vascular resistance in patients with idiopathic pulmonary arterial hypertension (IPAH). Increased Ca(2+) influx is an important stimulus for PASMC proliferation. Transient receptor potential (TRP) channel genes encode Ca(2+) channels that are responsible for Ca(2+) entry during cell proliferation. Normal human PASMC expressed multiple canonical TRP (TRPC) isoforms; TRPC6 was highly expressed and TRPC3 was minimally expressed. The protein expression of TRPC6 in normal PASMC closely correlated with the expression of Ki67, suggesting that TRPC6 expression is involved in the transition of PASMC from quiescent phase to mitosis. In lung tissues and PASMC from IPAH patients, the mRNA and protein expression of TRPC3 and -6 were much higher than in those from normotensive or secondary pulmonary hypertension patients. Inhibition of TRPC6 expression with TRPC6 small interfering RNA markedly attenuated IPAH-PASMC proliferation. These results demonstrate that expression of TRPC channels correlates with the progression of the cell cycle in PASMC. TRPC channel overexpression may be partially responsible for the increased PASMC proliferation and pulmonary vascular medial hypertrophy in IPAH patients.

Adult↗

Bosentan inhibits transient receptor potential channel expression in pulmonary vascular myocytes.

Bosentan, a dual endothelin receptor blocker, has been used clinically to treat idiopathic pulmonary arterial hypertension (IPAH). However, the mechanism of its antiproliferative effect on pulmonary artery smooth muscle cells (PASMCs) remains unclear. A rise in cytoplasmic Ca2+ stimulates PASMC proliferation and the canonical transient receptor potential (TRPC) channels are an important pathway for Ca2+ entry during PASMC proliferation. Bosentan (20-50 microM) significantly inhibited endothelin-1- or platelet-derived growth factor (PDGF)-mediated PASMC growth and [3H]thymidine uptake. In PASMCs, endothelin-1 (1 microM) and PDGF (10 ng/ml) both upregulated protein expression of TRPC6, whereas bosentan markedly downregulated TRPC6 protein levels. Furthermore, TRPC6 expression in PASMCs from patients with IPAH was greater than in normal PASMCs, and the antiproliferative effect of bosentan was significantly enhanced in IPAH-PASMCs in comparison with normal PASMCs. These observations demonstrate that the antiproliferative effect of bosentan on PASMCs involves the downregulation of TRPC6 channels via a mechanism possibly independent of endothelin receptor blockade. The greater effect of bosentan on IPAH-PASMCs than on normal PASMCs suggests that increased TRPC6 expression and function may be involved in the overgrowth of PASMCs in patients with IPAH.

Animals↗

Cre-mediated reversible immortalization of human renal proximal tubular epithelial cells.

Primary human renal proximal tubule epithelial cells (RPTECs) are of limited use for basic research and for clinical applications due to their limited lifespan in culture. Here we used two lentivirus vectors carrying the human telomerase (hTERT) and the SV40T antigen (Tag) flanked by loxP sites to reversibly immortalize RPTECs. Transduced RPTEC clones continued to proliferate while retaining biochemical and functional characteristics of primary cells. The clones exhibited contact-inhibited, anchorage- and growth factor-dependent growth and did not form tumors in nude mice, suggesting that the cells were not transformed. Transient Cre expression in these cells led to efficient proviral deletion, upregulation of some renal specific activities, and decreased growth rates. Ultimately, the cells underwent replicative senescence, indicating intact cell cycle control. Thus, reversible immortalization allows the expansion of human RPTECs, leading to large production of RPTECs that retain most tissue-specific properties.

Antigens, Polyomavirus Transforming↗

Successful correction of the human beta-thalassemia major phenotype using a lentiviral vector.

beta-thalassemias are the most common single gene disorders and are potentially amenable to gene therapy. However, retroviral vectors carrying the human beta-globin cassette have been notoriously unstable. Recently, considerable progress has been made using lentiviral vectors, which stably transmit the beta-globin expression cassette. Thus far, mouse studies have shown correction of the beta-thalassemia intermedia phenotype and a partial, variable correction of beta-thalassemia major phenotype. We tested a lentiviral vector carrying the human beta-globin expression cassette flanked by a chromatin insulator in transfusion-dependent human thalassemia major, where it would be ultimately relevant. We demonstrated that the vector expressed normal amounts of human beta-globin in erythroid cells produced in in vitro cultures for unilineage erythroid differentiation. There was restoration of effective erythropoiesis and reversal of the abnormally elevated apoptosis that characterizes beta-thalassemia. The gene-corrected human beta-thalassemia progenitor cells were transplanted into immune-deficient mice, where they underwent normal erythroid differentiation, expressed normal levels of human beta-globin, and displayed normal effective erythropoiesis 3 to 4 months after xenotransplantation. Variability of beta-globin expression in erythroid colonies derived in vitro or from xenograft bone marrow was similar to that seen in normal controls. Our results show genetic modification of primitive progenitor cells with correction of the human thalassemia major phenotype.

Animals↗

Cu(I)-mediated reductive amination of boronic acids with nitroso aromatics.

[reaction: see text] A mild method for the reductive amination of aryl boronic acids with nitroso aromatic compounds is reported. This C-N bond formation is mediated by a stoichiometric amount of CuCl as both a catalyst and a reducing agent. Alternatively, 10% Cu(I)-3-methylsalicylate (CuMeSal) catalyzes the same reaction in the presence of either ascorbic acid or hydroquinone as the terminal reducing agent. Diarylamines bearing a variety of functional groups can be obtained in good yields.

Amination↗

Overexpression of TRPC1 enhances pulmonary vasoconstriction induced by capacitative Ca2+ entry.

Transient receptor potential (TRP) cation channels are a critical pathway for Ca2+ entry during pulmonary artery (PA) smooth muscle contraction. However, whether canonical TRP (TRPC) subunits and which TRP channel isoforms are involved in store depletion-induced pulmonary vasoconstriction in vivo remain unclear. This study was designed to test whether overexpression of the human TRPC1 gene (hTRPC1) in rat PA enhances pulmonary vasoconstriction due to store depletion-mediated Ca2+ influx. The hTRPC1 was infected into rat PA rings with an adenoviral vector. RT-PCR and Western blot analyses confirmed the mRNA and protein expression of hTRPC1 in the arterial rings. The amplitude of active tension induced by 40 mM K+ (40K) in PA rings infected with an empty adenoviral vector (647 +/- 88 mg/mg) was similar to that in PA rings infected with hTRPC1 (703 +/- 123 mg/mg, P = 0.3). However, the active tension due to capacitative Ca2+ entry (CCE) induced by cyclopiazonic acid was significantly enhanced in PA rings overexpressing hTRPC1 (91 +/- 13% of 40K-induced contraction) compared with rings infected with an empty adenoviral vector (61 +/- 14%, P < 0.001). Endothelial expression of hTRPC1 was not involved since the CCE-induced vasoconstriction was also enhanced in endothelium-denuded PA rings infected with the adenoviral vector carrying hTRPC1. These observations demonstrate that hTRPC1 is an important Ca(2+)-permeable channel that mediates pulmonary vasoconstriction when PA smooth muscle cell intracellular Ca2+ stores are depleted.

Adenoviridae↗

E-selectin is required for the antiangiogenic activity of endostatin.

Endostatin, a 20-kDa fragment of collagen XVIII, is a potent angiogenesis inhibitor. E-selectin, an inducible leukocyte adhesion molecule specifically expressed by endothelial cells, has also been implicated in angiogenesis. By using in vivo, ex vivo, and in vitro angiogenic assays, we investigated the functional relationship between endostatin and E-selectin. In corneal micropocket assays, recombinant endostatin administered i.p. by osmotic pump inhibited basic fibroblast growth factor-induced angiogenesis in WT, but not E-selectin-deficient, mice. Similarly, endostatin inhibited vascular endothelial growth factor-stimulated endothelial sprout formation from aortic rings dissected from WT but not from E-selectin-deficient mice. To further explore this apparent requirement for E-selectin in endostatin action, we manipulated E-selectin expression in cultured human endothelial cells. When E-selectin was induced by IL-1beta, or lipopolysaccharide, human umbilical vein endothelial cells and human dermal microvascular endothelial cells each became markedly more sensitive to inhibition by endostatin in a vascular endothelial growth factor-induced cell migration assay. To dissociate E-selectin expression from other consequences of endothelial activation, human umbilical vein endothelial cells were transduced with an adenoviral human E-selectin expression construct; these cells also showed increased sensitivity to endostatin, and this effect required the E-selectin cytoplasmic domain. Taken together, these results indicate that E-selectin is required for the antiangiogenic activity of endostatin in vivo and ex vivo and confers endostatin sensitivity to nonresponsive human endothelial cells in vitro. E-selectin may be a useful predictor and modulator of endostatin efficacy in antiangiogenic therapy.

Angiogenesis Inhibitors↗

Copper-mediated, palladium-catalyzed coupling of thiol esters with aliphatic organoboron reagents.

Thiol esters and B-alkyl-9-BBN derivatives couple in the presence of a copper(I) carboxylate mediator and a palladium catalyst. In contrast to copper-mediated, palladium-catalyzed cross-couplings of thioorganics with boronic acids, the current coupling reaction of 9-BBN derivatives is facilitated by the addition of a base such as Cs(2)CO(3). Under optimized conditions, a variety of thiol esters react with different B-alkyl-9-BBN derivatives giving ketones in moderate to excellent yields.

Boron Compounds↗

Adsorption of water-soluble dyes onto modified resin.

Adsorption behavior of five kinds of water-soluble dyes onto functionalized resin NKY has been dynamically and thermodynamically investigated. The results showed that the adsorption rates of dye K-GN, K-2BP, KN-R, AAB and 2G are all controlled by liquid film diffusion and particle diffusion. Equilibrium adsorption data can be well described by three-parameter equation. The first layer adsorption is the physical enhanced by chemical effect but multilayer adsorption is typically physical adsorption. Standard free energy change DeltaG(0) and standard entropy change DeltaS(0) indicate the adsorption of the dyes onto NKY can occur spontaneously.

Adsorption↗

Genetic diversity and relationship of Yunnan native cattle breeds and introduced beef cattle breeds.

In this study, random amplified polymorphic DNA (RAPD) analysis was used to estimate genetic diversity and relationship in 134 samples belonging to two native cattle breeds from the Yunnan province of China (DeHong cattle and DiQing cattle) and four introduced beef cattle breeds (Brahman, Simmental, MurryGrey, and ShortHorn). Ten primers were used, and a total of 84 bands were scored, of which 63 bands (75.0%) were polymorphic. The genetic distance matrix was obtained by proportions of shared fragment. The results indicate that the Yunnnan DeHong cattle breed is closely related to the Brahman (Bos indicus), and the Yunnan DiQing cattle breed is closely related to the Simmental, ShortHorn, and MurryGrey (Bos taurus) breeds. Our results imply that Bos indicus and Bos taurus were the two main origins of Yunnan native cattle. The results also provide the basic genetic materials for conservation of cattle resources and crossbreeding of beef cattle breeds in South China.

Animals↗

Altered endochondral bone development in matrix metalloproteinase 13-deficient mice.

The assembly and degradation of extracellular matrix (ECM) molecules are crucial processes during bone development. In this study, we show that ECM remodeling is a critical rate-limiting step in endochondral bone formation. Matrix metalloproteinase (MMP) 13 (collagenase 3) is poised to play a crucial role in bone formation and remodeling because of its expression both in terminal hypertrophic chondrocytes in the growth plate and in osteoblasts. Moreover, a mutation in the human MMP13 gene causes the Missouri variant of spondyloepimetaphyseal dysplasia. Inactivation of Mmp13 in mice through homologous recombination led to abnormal skeletal growth plate development. Chondrocytes differentiated normally but their exit from the growth plate was delayed. The severity of the Mmp13- null growth plate phenotype increased until about 5 weeks and completely resolved by 12 weeks of age. Mmp13-null mice had increased trabecular bone, which persisted for months. Conditional inactivation of Mmp13 in chondrocytes and osteoblasts showed that increases in trabecular bone occur independently of the improper cartilage ECM degradation caused by Mmp13 deficiency in late hypertrophic chondrocytes. Our studies identified the two major components of the cartilage ECM, collagen type II and aggrecan, as in vivo substrates for MMP13. We found that degradation of cartilage collagen and aggrecan is a coordinated process in which MMP13 works synergistically with MMP9. Mice lacking both MMP13 and MMP9 had severely impaired endochondral bone, characterized by diminished ECM remodeling, prolonged chondrocyte survival, delayed vascular recruitment and defective trabecular bone formation (resulting in drastically shortened bones). These data support the hypothesis that proper ECM remodeling is the dominant rate-limiting process for programmed cell death, angiogenesis and osteoblast recruitment during normal skeletal morphogenesis.

Animals↗

[Investigation of the influence of nitric oxide on the survival of a random pattern skin flap].

OBJECTIVE: To investigate the changes of nitric oxide concentration in the distal portion of a random pattern skin flap and the influence of the exogenous L-arginine on the survival of the random pattern skin flap. METHODS: A random pattern skin flap (7 cm x 2 cm) was cranially designed and elevated on the back of a Wistar rat. An image analysis technology was used to evaluate the survival rate of the skin flap, while a biochemistry method was used to test the concentrations of the NO in the tissue. RESULTS: The survival area of the flap in the L-arginine-treated group was significantly enlarged (63.83 +/- 5.13)% (P < 0.01) in seven days postoperatively, compared with the control group (43.26 +/- 2.86)%. The NO concentration in the tissue was no statistic difference between all of the groups immediately after the operation (P > 0.05). But, the NO concentration in the control was decreasing at the beginning and then increasing slightly to reach the high level in 12 hours after the operation. It was thereafter slumped down to the baseline in 72 hours after the surgery. Although the changes in the L-arginine-treated group were quite similar to the control excepting of the extent, the NO concentration was kept in a higher level in the sequential time after the operation (P < 0.01, P < 0.01, P < 0.05 and P < 0.01). CONCLUSIONS: The NO concentration in skin flap tissue after the elevation was going up slightly for a short time. The exogenous L-arginine could promote the NO concentration in the random pattern skin flap to protect it from ischemic injury.

Animals↗

[Gene differential expression of liver tissues in crossbred versus purebred chicken and their relationship with heterosis of meat trait].

The concept of heterosis has already been put forward for a century. The hypothesis of Dominance, Superdominace and Epistasis has also been brought forward to explain the phenomenon of heterosis. As we know, there is spatio-temporal speciality about the expression of gene and only expressed genes contribute to the formation of heterosis. So the study on heterosis in expression level becomes more meaningful. A lot of studies on heterosis in this level have been done in plants, but there is no such study carried on animals in this area. In this study, the technique of mRNA Reverse Transcription Differential Display was used to research the heterosis molecular mechanism of animal. In order to expound the molecular genetic mechanism of animals heterosis, the 4 x 4 completely diallele cross experiment of 4 purebreds chicken was conducted among White Polymouth Rock (EE), Chinese Silk Chicken (CC), CAU Brown (DD) and White Leghorn (AA). The chicken of 16 cross combinations were reared to 8 weeks old, then 30 chicken in each combination were selected randomly and slaughtered. The traits of body weight of 8 weeks, wing weight, eviscerated weight, eviscerated weight with giblet, breast muscle yield, leg muscle yield, body length, abdomen fat weight, intramuscular fat width, tibia length were measured, and in which 8 individuals in each combination were selected randomly to collect the liver tissue samples, which were stored in liquid nitrogen or at -80 degrees C to be used for total RNA (TRNA) extracting. After the total RNA (TRNA) was extracted, 16 TRNA pools were formed in the same quantitative according to the concentration of 8 individual TRNA. They were reversely transcribed with three anchor primers H-T11 A, H-T11 G and H-T11 C. Then the reverse transcription PCR for each transcript product was done in two repeats at the same time with the same anchor primers and 8 random primers. The polyacrylamide gel electrophoresis of each PCR product was run in Bio-Rad Power 3,000 temperature control system. After electrophoresis, the gel was stained by AgNO3 according to the stain method described by Echt et al. The differential display bands in the polyacrylamide gel were counted. The band displayed is counted as 1 whereas no band is counted as 0 and only expression bands reproducible in two repeats were statistically analyzed. The correlation analysis between heterosis percentage and gene expression patterns was done with statistic analysis software (SAS) package. The statistic results indicated that among 690 total numbers of bands, the percentage of differential expression bands reproducible (457) is 66.23%. Eight kinds of gene differential expression patterns were found and listed as follow: 1): Band presents only in one purebred (P1);2): Band in one crossbred and its corresponding paternal purebred; or Band in one crossbred and its corresponding maternal purebred (P2);3): Band in purebreds and one crossbred (P3); 4): Band only in one crossbred (P4);5): Bands in both crossbreds and one purebred (P5);6): Bands only in both crossbreds (P6);7): Bands only in purebreds (P7);8): Bands both in purebreds and crossbreds P8. The differential expression of gene between purebred and crossbred chicken was detected for the first time. The proportion of each pattern in each kind of purebred combination is different. The percentage of P8 (75.34%) is the highest. The total percentage of differential expression patterns (24.66%) showed that the gene differential expression exists as a matter of fact. Among all the gene differential expression patterns, the percentage of P3 is the highest whereas the percentage of P7, P6 and P4 is very low, it indicated that different genes may have different expression patterns in purebreds and crossbreds. The results are similar to the study results on plants, which indicates that the gene differential expression between purebred and crossbred exists universally in biology. The correlation between gene expression patterns and heterosis percentage was studied, but correlation between P8 and the heterosis percentage is not significant (P > 0.05), it indicates that some patterns of gene differential expression may be the molecular genetic basic of heterosis. Among all the gene differential expression patterns, each pattern affects the expression of meat trait in different manner. There is significantly negative correlation between P4 and heterosis percentage of body weight of 8 weeks, breast muscle yield, leg muscle yield, eviscerated weight with giblet and eviscerated weight (P < 0.05); P1 is of significantly negative correlation with heterosis percentage of abdomen fat weight (P < 0.05) and of very significantly negative correlation with heterosis percentage of body length (P < 0.01); The negative correlation between P2 and heterosis percentage of intramuscular fat width is significant (P < 0.05); The positive correlation between P7 and heterosis percentage of leg muscle yield, wing weight, eviscerated weight with giblet and intramuscular fat width is significant (P < 0.05); The positive correlation between P5 and heterosis percentage of tibia length (P < 0.05) is significant. These results show that these 5 kinds of patterns play important role in heterosis forming of meat trait. P1 and P7 show that expressed gene in purebreds is depressed; P4 indicates that new gene expression occurs in crossbreds; P5 reveals that expressed gene only in one purebred express in all crossbreds. All genes of crossbreds come from purebred, which are not only the simple adding of these purebred genes, giving birth to unknown interaction between these genes coming from different purebreds, then leading to differential expression of genes. These gene differential expressions maybe form the heterosis of meat trait.

Animals↗

Intramural pregnancy: a report of 2 cases.

BACKGROUND: Intramural pregnancy is a rare type of ectopic pregnancy and may be easily misdiagnosed as cornual pregnancy or trophoblastic tumor. Hysterectomy is performed due to extensive bleeding and uterine rupture in most cases. The incidence is <1% of ectopic pregnancy. Eighteen cases were reported in the People's Republic of China and 33 in the rest of the world since 1957. For a young woman who wishes to maintain her fertility, it is important to make an early diagnosis and to undertake conservative treatment. CASES: A 29-year-old woman, gravida 3, para 1, was admitted because of missed periods for >70 days, 1 week of mild vaginal bleeding and lower abdominal pain. Her serum beta-human chorionic gonadotropin (beta-hCG) level was 765 U/L. Transvaginal sonography (TVS) revealed an ill-defined mass measuring 3.0 x 3.5 x 2.0 cm within the fundal myometrium adjacent to the covering. At laparotomy, a mass 3 cm in diameter bulged from the left fundal covering and was resected to the surface of myometrium. The patient preserved her fertility through successful repair of the uterus. A 39-year-old woman, gravida 4, para 1, was admitted because of irregular vaginal bleeding for 2 months after intrauterine device insertion. Her serum beta-hCG level was 228 U/L. TVS revealed amorphous echoes in the uterine cavity. We made an initial, presumptive diagnosis of incomplete abortion. Curettage was performed, but no fetal elements were found. The serum beta-hCG level was 360 U/L after 1 week. Computed tomography revealed a trophoblastic tumor with deep invasion of the myometrium. A subradical abdominal hysterectomy was performed and gave the impression of chorionic carcinoma. Pathologic examination revealed diffuse hemorrhage and early invasion of chorionic villi in the fundal myometrium with focal decidual reaction of the endometrium. The diagnosis of intramural pregnancy was made after the operation. CONCLUSION: Both cases of intramural pregnancy were treated successfully.

Abdominal Pain↗

[Comparison on desorptive behavior of copper in phaeozem and burozem].

Desorptive behavior of Cu2+ from phaeozem and burozem was studied and compared using the batch method. The results indicated that Cu2+ desorbed from phaeozem tested had a longer lag period than that from burozem. When adsorbed Cu2+ for phaeozem was lower than 729.00 mg x kg(-1) (Cu2+ concentration was lower than 40 mg x L(-1)), and on burozem was lower than 393.29 mg x kg(-1) (Cu2+ concentration was lower than 20 mg x L(-1)), adsorbed Cu2+ was hardly desorbed. Meanwhile, the desorption ratio of Cu2+ from the two soils were both low, especially for phaeozem. In the range of Cu2+ concentration used in the experiments, the desorption rate maximums of Cu2+ for phaeozem and burozem were respectively 6.60% and 13.56%. Freundlich equation could best describe desorption isotherm processes of Cu2+ from the two soils. Adsorbed Cu2+ on soils was desorbed so fast from phaeozem and burozem that the desorption equilibrium could be reached only after 40 min desorption. The best model to describe Cu2+ desorption kinetics was two-constant equations, followed by the Elovich equation.

Adsorption↗

[Differential expression of CIDH gene between purebreds and hybrids in chicken and relationship with heterosis].

The technique of silver staining mRNA Differential Display was used to check the molecular genetic mechanism of heterosis in chicken. A differentially expressed fragment CE15A15, which was only expressed in two crossbreds, was found between purebreds and crossbreds. It is proved that the fragment intensively expresses in crossbreds through reverse Northern dot blot and RT-PCR. It shares 97% of sequence identity with a partial cDNA sequence of chicken that has been deposited in GenBank (AW198493) and believed to be identity with the cytosolic isocitrate dehydrogenase (CIDH) gene (ID:AF048831) of Microtus mexicanus. This suggests that the identified fragment may be a partial cDNA sequence of CIDH gene. The statistic analyses of phenotypes showed that the body weight and the carcass traits in crossbred of Silk Chicken (CC) x White Polymouth Rock (EE) and White Polymouth Rock (EE) x Silk Chicken (CC) had negative heterosis, while their fatty traits had positive heterosis. CIDH directly participates in the biosynthesis of fatty acids. This indicates that differential expression of the fragment between purebreds and crossbreds might be closely related to heterosis of crossbreds.

Animals↗

Endothelial progenitor cells in infantile hemangioma.

Infantile hemangioma is an endothelial tumor that grows rapidly after birth but slowly regresses during early childhood. Initial proliferation of hemangioma is characterized by clonal expansion of endothelial cells (ECs) and neovascularization. Here, we demonstrated mRNA encoding CD133-2, an important marker for endothelial progenitor cells (EPCs), predominantly in proliferating but not involuting or involuted hemangioma. Progenitor cells coexpressing CD133 and CD34 were detected by flow cytometry in 11 of 12 proliferating hemangioma specimens from children 3 to 24 months of age. Furthermore, in 4 proliferating hemangiomas, we showed that 0.14% to 1.6% of CD45(-) nucleated cells were EPCs that coexpressed CD133 and the EC marker KDR. This finding is consistent with the presence of KDR(+) immature ECs in proliferating hemangioma. Our results suggest that EPCs contribute to the early growth of hemangioma. To our knowledge, this is the first study to show direct evidence of EPCs in a human vascular tumor.

AC133 Antigen↗

Toxicity and repellency of patchouli oil and patchouli alcohol against Formosan subterranean termites Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae).

Patchouli oil obtained from Pogostemon cablin (Blanco) Benth and its main constituent, patchouli alcohol, were tested for their repellency and toxicity against Formosan subterranean termites (Coptotermes formosanus Shiraki). Both were found to be toxic and repellent. Unusual tissue destruction was noted inside the exoskeleton of the termite after patchouli alcohol was topically applied to the dorsum.

Animals↗