Cytoprotective Capacity Research Articles

OCTN1‐Mediated Ergothioneine Uptake Protects Lung Epithelial Cells From Tobacco Smoke‐Induced Damage

Smoking is a major contributing factor for chronic obstructive pulmonary disease (COPD). Tobacco smoke exposure causes oxidative stress which results in alveolar wall destruction, mucus hyper secretion, inflammation and defective tissue repair. Ergothioneine (ET), an amino acid transported by OCTN1, has been shown to exhibit antioxidant and cytoprotective capacities in vitro. Hence, the objective of this study is to understand the role of OCTN1 and the effect of ET on human alveolar cells when subjected to tobacco smoke‐induced oxidative stress.Confocal microscopy and Western blot was used to determine the presence of OCTN1 in A549 cells and freshly isolated human alveolar epithelial type II (ATII) and type I‐like (ATI‐like) cells in primary culture. A549 cells were cultured in medium containing 1 mM of ET and its intracellular accumulation was measured by LC‐MS/MS over a 3‐week period. A549 cells were also was exposed to various concentrations of cigarette smoke extract (CSE). Prior to the experiment, cells were either cultured for 1 week in the presence or absence of 1 mM ET. An MTT assay was used to assess cell viability for the difference exposures to CSE.Immunoblot and CLSM showed OCTN1 expression in A549, ATII and ATI‐like cells. The uptake of ET by A549 epithelial cells was saturated after approximately 7 days of culture. Western blot data supported the CLSM results and also indicated that OCTN1 levels were increased in the presence of ET. The MTT assay showed that cells pre‐treated with ET were less affected by the cytotoxic effects of CSE.These preliminary data indicate that OCTN1 might play a role in the pathogenesis of tobacco smoke‐induced COPD by regulating ergothioneine transport.

Erythropoietin and diabetes mellitus.

Erythropoietin (EPO) is a 30.4 kDa growth factor and cytokine that governs cell proliferation, immune modulation, metabolic homeostasis, vascular function, and cytoprotection. EPO is under investigation for the treatment of variety of diseases, but appears especially suited for the treatment of disorders of metabolism that include diabetes mellitus (DM). DM and the complications of this disease impact a significant portion of the global population leading to disability and death with currently limited therapeutic options. In addition to its utility for the treatment of anemia, EPO can improve cardiac function, reduce fatigue, and improve cognition in patients with DM as well as regulate cellular energy metabolism, obesity, tissue repair and regeneration, apoptosis, and autophagy in experimental models of DM. Yet, EPO can have adverse effects that involve the vasculature system and unchecked cellular proliferation. Critical to the cytoprotective capacity and the potential for a positive clinical outcome with EPO are the control of signal transduction pathways that include protein kinase B, the mechanistic target of rapamycin, Wnt signaling, mammalian forkhead transcription factors of the O class, silent mating type information regulation 2 homolog 1 (Saccharomyces cerevisiae), and AMP activated protein kinase. Therapeutic strategies that can specifically target and control EPO and its signaling pathways hold great promise for the development of new and effective clinical treatments for DM and the complications of this disorder.

Renal Integrin-Linked Kinase Depletion Induces Kidney cGMP-Axis Upregulation: Consequences on Basal and Acutely Damaged Renal Function.

Soluble guanylyl cyclase (sGC) is activated by nitric oxide (NO) and produces cGMP, which activates cGMP-dependent protein kinases (PKG) and is hydrolyzed by specific phosphodiesterases (PDE). The vasodilatory and cytoprotective capacity of cGMP-axis activation results in a therapeutic strategy for several pathologies. Integrin-linked kinase (ILK), a major scaffold protein between the extracellular matrix and intracellular signaling pathways, may modulate the expression and functionality of the cGMP-axis-related proteins. We introduce ILK as a novel modulator in renal homeostasis as well as a potential target for cisplatin (CIS)-induced acute kidney injury (AKI) improvement. We used an adult mice model of depletion of ILK (cKD-ILK), which showed basal increase of sGC and PKG expressions and activities in renal cortex when compared with wildtype (WT) littermates. Twenty-four h activation of sGC activation with NO enhanced the filtration rate in cKD-ILK. During AKI, cKD-ILK maintained the cGMP-axis upregulation with consequent filtration rates enhancement and ameliorated CIS-dependent tubular epithelial-to-mesenchymal transition and inflammation and markers. To emphasize the role of cGMP-axis upregulation due to ILK depletion, we modulated the cGMP axis under AKI in vivo and in renal cultured cells. A suboptimal dose of the PDE inhibitor ZAP enhanced the beneficial effects of the ILK depletion in AKI mice. On the other hand, CIS increased contractility-related events in cultured glomerular mesangial cells and necrosis rates in cultured tubular cells; ILK depletion protected the cells while sGC blockade with ODQ fully recovered the damage.

Use of in vitro human keratinocyte models to study the effect of cooling on chemotherapy drug-induced cytotoxicity

A highly distressing side-effect of cancer chemotherapy is chemotherapy-induced alopecia (CIA). Scalp cooling remains the only treatment for CIA, yet there is no experimental evidence to support the cytoprotective capacity of cooling. We have established a series of in vitro models for the culture of human keratinocytes under conditions where they adopt a basal, highly-proliferative phenotype thus resembling the rapidly-dividing sub-population of native hair-matrix keratinocytes. Using a panel of chemotherapy drugs routinely used clinically (docetaxel, doxorubicin and the active metabolite of cyclophosphamide 4-OH-CP), we demonstrate that although these drugs are highly-cytotoxic, cooling can markedly reduce or completely inhibit drug cytotoxicity, in agreement with clinical observations. By contrast, we show that cytotoxicity caused by specific combinatorial drug treatments cannot be adequately attenuated by cooling, supporting data showing that such treatments do not always respond well to cooling clinically. Importantly, we provide evidence that the choice of temperature may be critical in determining the efficacy of cooling in rescuing cells from drug-mediated toxicity. Therefore, despite their reductive nature, these in vitro models have provided experimental evidence for the clinically-reported cytoprotective role of cooling and represent useful tools for future studies on the molecular mechanisms of cooling-mediated cytoprotection.

Chemical and cellular antioxidative properties of threadfin bream (Nemipterus spp.) surimi byproduct hydrolysates fractionated by ultrafiltration

Protein hydrolysate from frame, bone and skin (FBSH) of threadfin bream was prepared using Virgibacillus sp. SK33 proteinase and fractionated using sequential ultrafiltration membranes with molecular weight cut-offs (MWCO) of 30, 5 and 1kDa, respectively. Four fractions, namely FBSH-I (>30kDa), FBSH-II (5–30kDa), FBSH-III (1–5kDa), and FBSH-IV (<1kDa), were obtained. All fractions were rich in Lys, Glu/Gln, Gly, Pro, Ala, Asp/Asn, and Arg. FBSH-III and FBSH-IV showed the highest surface hydrophobicity measured by 8-anilino-1-naphthalenesulfonic acid (ANS) probe (p<0.05). FBSH-III showed the highest antioxidant activity and cytoprotective effects against tert-butyl hydroperoxide (TBHP)-induced cytotoxicity of Caco-2 cells. In addition, FBSH-III inhibited lactate dehydrogenase (LDH) leakage and intracellular reactive species (ROS) production in a dose-dependent manner. FBSH-III retained antioxidant activity and cytoprotective capacity after in vitro simulated gastrointestinal digestion. These results suggested that FBSH-III might potentially be nutraceutical peptides with antioxidative properties.

The antioxidant activity of soursop decreases the expression of a member of the NADPH oxidase family

Cellular oxidative stress produced by an increase in free radicals is one of the factors that promote the development of chronic degenerative diseases; therefore, consuming natural antioxidants helps minimize their negative effects. This study evaluated the cytotoxicity of the soursop extract (Annona muricata), its cytoprotective capacity against oxidative stress induced by hydrogen peroxide, the inhibitory potential of reactive oxygen species (ROS), the molecular mechanism of its antioxidant action, and its capacity to repair cellular damage in the fibroblast cell line. The soursop extract proved not to be cytotoxic in fibroblast cultures and showed cytoprotective capacity against hydrogen peroxide-induced stress; in cell culture it reduced the generation of ROS significantly by inhibiting a sub-unit of the NADPH oxidase enzyme (p47phox). The soursop extract can prevent damage caused by cellular oxidants.

PSS270 - Reactivity and Cytoprotective Capacity of the Synthetic Catalytic Antioxidants Mnporphyrins towards Peroxynitrite and Hypochlorite

PSS270 - Reactivity and Cytoprotective Capacity of the Synthetic Catalytic Antioxidants Mnporphyrins towards Peroxynitrite and Hypochlorite

Dehydroepiandrosterone Increases Pancreatic Duodenal Homebox-1 (PDX-1) and Reduces Cleaved Caspase-3 Protein Expression in Insulin-Secreting INS-1E Cells

Due to decreases in dehydroepiandrosterone (DHEA) levels in an age group where there is increased incidence of chronic degenerative diseases, like diabetes mellitus type 2, we sought to identify the effect of incubation with DHEA on insulin secretory and cytoprotective capacity in pancreatic beta cells. The INS-1E cells were incubated in different concentrations of DHEA (10 µM, 100 nM and 1 nM) or vehicle (dimethyl sulfoxide; DMSO) for 1 or 7 days. Insulin secretion stimulated by glucose (2.8 mM and 16.7 mM), determination of superoxide content, and the protein content of insulin receptor substrate-2 (IRS-2), extracellular regulated kinase 1 and 2 (ERK ½), phosphorylated extracellular kinase 1 and 2 (pERK ½), phosphorylated c-Jun Nterminal kinase (pJNK), cleaved caspase-3 and pancreatic duodenal homebox-1 (PDX-1) were analyzed. We observed no effects of DHEA incubation on insulin secretion. In contrast, DHEA incubation improved total insulin content. Moreover, we have observed no influence on the production of superoxide. In addition, an increased PDX-1 and reduced cleaved caspase-3 concentration were observed. Our data point out to a stimulatory action of DHEA on insulin production and cytoprotective effects.

The tellurium redox immunomodulating compound AS101 inhibits IL-1β-activated inflammation in the human retinal pigment epithelium

PurposeAS101 is a non-toxic organotellurium-IV compound with demonstrated immunomodulating activity in vitro and in vivo. Inflammatory responses are attributed to the pathophysiology of numerous ocular diseases. In this study, we...

Autophagy Attenuates Diabetic Glomerular Damage through Protection of Hyperglycemia-Induced Podocyte Injury

Despite the recent attention focused on the important role of autophagy in maintaining podocyte homeostasis, little is known about the changes and mechanisms of autophagy in podocyte dysfunction under diabetic condition. In this study, we investigated the role of autophagy in podocyte biology and its involvement in the pathogenesis of diabetic nephropathy. Podocytes had a high basal level of autophagy. And basal autophagy inhibition either by 3-methyladenenine (3-MA) or by Beclin-1 siRNA was detrimental to its architectural structure. However, under diabetic condition in vivo and under high glucose conditions in vitro, high basal level of autophagy in podocytes became defective and defective autophagy facilitated the podocyte injury. Since the dynamics of endoplasmic reticulum(ER) seemed to play a vital role in regulating the autophagic flux, the results that Salubrinal/Tauroursodeoxycholic acid (TUDCA) could restore defective autophagy further indicated that the evolution of autophagy may be mediated by the changes of cytoprotective output in the ER stress. Finally, we demonstrated in vivo that the autophagy of podocyte was inhibited under diabetic status and TUDCA could improve defective autophagy. Taken together, these data suggested that autophagy might be interrupted due to the failure of ER cytoprotective capacity upon high glucose induced unmitigated stress, and the defective autophagy might accelerate the irreparable progression of diabetic nephropathy.

Effects of Everolimus on Oxidative Stress in Kidney Model of Ischemia/Reperfusion Injury

Background/Aims: Reactive oxygen species play an important role in the pathogenesis of kidney ischemia/reperfusion injury (IRI) which may be influenced by immunosuppressive therapy. Pertinent to this, we investigated the effects of the mTOR inhibitor everolimus on redox settings and the activity of the anti-oxidative system in kidneys exposed to IRI. Methods: C57BL/6 mice were subjected to IRI by clamping both renal pedicles for 45 min. Everolimus was applied in daily, subcutaneous doses (0.25 mg/kg body weight), starting 1 day before IRI induction. Both everolimus-treated and non-treated mice were sacrificed at several time points, starting 30 min and finishing 7 days after IRI induction. Markers of oxidation such as glutathione and NADPH levels and anti-oxidative enzyme activities were determined in the kidneys. Results: In comparison to both sham and non-treated animals, the treatment with everolimus resulted in an increased level of markers of oxidation, including a lower level of glutathione, increased level of oxidized glutathione and reduced level of NADPH. The activity of superoxide dismutase was reduced in both experimental groups, but the effects were less pronounced in everolimus-treated animals. In the early phase of reperfusion, everolimus-treated animals showed higher activity of glutathione reductase in comparison to non-treated animals, whereas the activities of glutathione peroxidase and catalase were generally similar. The treatment with everolimus significantly reduced heme oxygenase-1 expression and increased iNOS mRNA expression when compared to non-treated animals. Conclusion: Our data imply that everolimus treatment may decrease cytoprotective capacity in kidneys exposed to IRI due to promoted oxidative/nitrosative stress.

ARA290, a non-erythropoietic EPO derivative, attenuates renal ischemia/reperfusion injury

BackgroundIn contrast with various pre-clinical studies, recent clinical trials suggest that high dose erythropoietin (EPO) treatment following kidney transplantation does not improve short-term outcome and that it even increases the risk of thrombotic events. ARA290 is a non-erythropoietic EPO derivative and does not increase the risk of cardiovascular events, but potentially has cytoprotective capacities in prevention of renal ischemia/reperfusion injury.MethodsEight female Dutch Landrace pigs were exposed to unilateral renal ischemia for 45 minutes with simultaneous cannulation of the ureter of the ischemic kidney. ARA290 or saline was administered by an intravenous injection at 0, 2, 4 and 6 hours post-reperfusion. The animals were sacrificed seven days post-reperfusion.ResultsARA290 increased glomerular filtration rate during the observation period of seven days. Furthermore, ARA290 tended to reduce MCP-1 and IL-6 expression 15 minutes post-reperfusion. Seven days post-reperfusion ARA290 reduced interstitial fibrosis.ConclusionsThe improvement in renal function following renal ischemia/reperfusion and reduced structural damage observed in this study by ARA290 warrants further investigation towards clinical application.

CDDO-9,11-dihydro-trifluoroethyl amide (CDDO-dhTFEA) induces hepatic cytoprotective genes and increases bile flow in rats

1. The transcription factor Nrf2 is important for hepatoprotection against oxidative stress, as it regulates many cytoprotective genes, including several important for glutathione (GSH) homeostasis. In addition to being an important endogenous antioxidant, GSH is also critical for the maintenance of bile acid-independent bile flow. While it has been well-established that synthetic oleanane triterpenoids pharmacologically activate Nrf2, their effects on bile flow and hepatic cytoprotective capacity have not been fully explored.2. The present studies were conducted to evaluate the effects of a compound in this class, CDDO-9,11-dihydro-trifluoroethyl amide (CDDO-dhTFEA), on these parameters. CDDO-dhTFEA at 3, 10 or 30 mg/kg was orally administered to bile duct-cannulated rats once daily for 7 days, with bile collected 5 h after each dose for 1 h. Livers were harvested after the final bile collection for the evaluation of histology and Nrf2 targets.3. CDDO-dhTFEA did not affect liver histology. CDDO-dhTFEA markedly and dose-dependently increased bile flow, as well as the biliary excretion of GSH, cholesterol and phospholipids without affecting biliary excretion of bile acids. This was accompanied by dose-dependent increases in mRNA expression and/or enzyme activity of a broad panel of cytoprotective Nrf2 target genes, including NAD(P)H quinone oxidoreductase 1 (Nqo1), thioredoxin reductase (Txnrd), sulfiredoxin 1(Srxn1), glutamate cysteine ligase catalytic and modifier subunits (Gclc and Gclm), glutathione reductase (Gsr), gamma-glutamyl transpeptidase 1 (Ggt1), heme oxygenase-1 (Ho-1) and epoxide hydrolase-1 (Eh-1).4. These data further demonstrate the important hepatobiliary attributes of oleanane synthetic triterpenoids and support their continued investigation for liver diseases.

Quantitative Histologic Evidence of Amifostine-Induced Cytoprotection in an Irradiated Murine Model of Mandibular Distraction Osteogenesis

Head and neck cancer management requires adjuvant radiotherapy. The authors have previously demonstrated the damaging effect of a human equivalent dose of radiation on a murine mandibular model of distraction osteogenesis. Using quantitative histomorphometry, the authors' specific aim was to objectively measure amifostine radioprotection of the cellular integrity and tissue quality of an irradiated and distracted regenerate. Sprague-Dawley rats were assigned randomly into two groups: radiotherapy/distraction osteogenesis and amifostine/radiotherapy/distraction osteogenesis, which received amifostine before radiotherapy. Both groups received a fractionated human equivalent dose of radiation prior to left mandibular osteotomy with fixator placement. Distraction to 5.1 mm was followed by a 28-day consolidation period. Quantitative histomorphometry was performed on left hemimandibles for osteocytes, empty lacunae, bone volume-to-tissue volume ratio, and osteoid volume-to-tissue volume ratio. Amifostine/radiotherapy/distraction osteogenesis exhibited bony bridging as opposed to radiotherapy/distraction osteogenesis fibrous unions. Quantitative histomorphometry analysis revealed statistically significant higher osteocyte count and bone volume-to-tissue volume ratio in amifostine-treated mandibles compared with irradiated mandibles. There was a corresponding decrease in empty lacunae and the ratio of osteoid volume-to-tissue volume between both groups. The authors have successfully established the significant osseous cytoprotective and histoprotective capacity of amifostine for distraction osteogenesis in the face of radiotherapy. The amifostine-sparing effect on bone cellularity correlated with increased bony unions and elimination of fibrous union. The authors posit that the demonstration of similar efficacy of amifostine in the clinic may allow the successful implementation of distraction osteogenesis as a viable reconstructive option for head and neck cancer in the future.

Statins inhibit iNOS‐mediated microbicidal potential of activated monocyte‐derived dendritic cells by an IFN‐β‐dependent mechanism

Statins are prescribed to 25 million people worldwide for treating hypercholesterolemia and reducing the risk of cardiovascular diseases. However, the side effects of statins on immunity, and particularly on DC immunobiology, have not been analyzed in-depth. Here, we have investigated the impact of lovastatin treatment during monocyte differentiation into DCs on the responsiveness of the resulting monocyte-derived DCs (moDCs) to TLR-mediated activation. Lovastatin positively regulated TLR4 signaling in LPS-stimulated moDCs, leading to strong activation of p38 MAP-kinase paralleled by increased proinflammatory cytokine and IFN-β production. In contrast, lovastatin promoted negative regulation of IFN-β-mediated autocrine signaling through the IFN-αβ receptor, paralleled by low expression of the transcription factor IRF-1, leading to the inhibition of the enzymes iNOS and HO-1. Defective activation of iNOS/HO-1 resulted in limited cytoprotective capacity against ROS and reduced microbicidal potential. These data were validated using an in vivo model of Listeria monocytogenes infection, which revealed that iNOS activation by splenic inflammatory moDCs, specialized in NO and TNF-α production, was strongly reduced in lovastatin-treated, Listeria-infected mice. Statin treatment could have severe implications in immunity against pathogens due to defective iNOS/HO-1 metabolism activation in inflammatory moDCs that might lead to immune failure.

Carboxy alkyl esters of Uncaria tomentosa augment recovery of sensorineural functions following noise injury

This study tested the hypothesis that hydrophilic chemotypes of the medicinal vine Uncaria tomentosa ( UT) would facilitate recovery of sensorineural functions following exposure to a damaging level of noise. The particular chemotypes investigated were carboxy alkyl esters (CAE) which are known to exhibit multifunctional cytoprotective properties that include: enhanced cellular DNA repair, antioxidation and anti-inflammation. Long–Evans rats were divided into four treatment groups: vehicle-control, noise-only, CAE-only and CAE + noise. The noise exposure was an 8 kHz octave band of noise at 105 dB SPL for 4 h. Outer hair cell (OHC) function was measured with the cubic 2 f 1– f 2 distortion product otoacoustic emissions (DPOAE) at the start of the study (baseline) and at time-points that corresponded to 1 day, 1 week and 4 weeks post-noise exposure to determine within-group effects. Compound action potentials to puretone stimuli were recorded from the VIIIth craniofacial nerve at 4 weeks post-noise exposure to determine between-group effects. Additionally, cytocochleograms were constructed for each row of OHCs from each group. Noise exposure produced significant sensorineural impairments. However, CAE treatment facilitated almost complete recovery of OHC function and limited the magnitude of cell loss. The loss of neural sensitivity to puretone stimuli was inhibited with CAE treatment. Therefore, it appears that the multifunctional cytoprotective capacity of CAE from UT may generalize to otoprotection from acoustic over-exposure.

Efecto citoprotector del camu-camu Myrciaria dubia en tres líneas celulares de ratón expuestos in vivo a bromato de potasio

It was evaluated in vivo the cytoprotective capacity of the fruit of Myrciaria dubia H.B.K. MC VAUGH "Camu- camu (Myrtacea) against mutagenic damage caused by potassium bromate (68.5 mg/k) on three mouse cell lines (liver, kidney and blood cells). Mice (n = 120) were divided into three groups which drank ad libitum: distilled water; TI (negative control) and TIII (positive control), the TII group (positive control) drank the aque- ous extract (2 %) of the fruit of Camu-camu. After ten days, only the TII and TIII groups were intraperitoneally injected with a single dose of KBr03(68.5 mg/k). The camu-camu treatment lasted 35 days more, where they were euthanized to determine the frequency of DNA damage by means of the alkaline comet assay protocol. It was observed in all cell lines a cytoprotective effect of camu-camu (p&lt;0.05) with respect with the negative control. The DNA-damaging effects of oxidative KBrO3 action is inhibited by the 2% aqueous extract of camu- camu fruit, probably by the presence of antioxidants such as ascorbic acid and flavonoids.

Therapeutic window for ferulic acid protection against noise-induced hearing loss in the guinea pig

Conclusion: Our results are in agreement with the general idea that natural antioxidants achieve their best cytoprotective capacity if given before and soon after the stressor. Objective: We focused on ferulic acid (FA, 4-hydroxy 3-methoxycinnamic acid), a phenolic compound that is known to exhibit antioxidant properties. Our study was designed to evaluate the effectiveness of FA for different schedules of treatment to establish the ‘therapeutic window’ for FA protection. Methods: Guinea pigs were exposed to acoustic trauma (6 kHz at 120 dB for 60 min) and received a total dose of 600 mg/kg of FA. Group I, noise control; group II, noise + FA (150 mg/kg) for 4 days starting 24 h post exposure; group III, noise + FA (60 mg/kg) 1 h before and 9 days post exposure; group IV, noise + FA (60 mg/kg) given 3 days before and 7 days post exposure; group V, noise + FA (150 mg/kg) 1 h before and 3 days post noise exposure. Auditory brainstem response (ABR) test and immunohistochemical and morphological studies were performed. Results: Group V had significantly decreased noise-induced hearing loss at day 21 from noise exposure. The improvement of auditory function by FA was paralleled by a significant reduction in oxidative stress marker. The other schedules of drug administration showed a minor degree of protection.

Adenovirus-Mediated Gene Expression of the Human c-FLIPL Gene Protects Pig Islets Against Human CD8 + Cytotoxic T Lymphocyte-Mediated Cytotoxicity

Cell-mediated immunity, especially of human CD8+ cytotoxic T lymphocytes (CTLs) is believed to have an important role in the long-term survival of pig islet xenografts. Protection against human CD8+ CTL cytotoxicity may reduce the direct damage to pig islets and enable long-term xenograft survival in pig-to-human islet xenotransplantation. We have previously reported that c-FLIP S/L genes, which are potent inhibitors of death receptor–mediated proapoptotic signals through binding competition with caspase-8 for recruitment to the Fas-associated via death domain (FADD), markedly suppress human CD8+ CTL-mediated xenocytotoxicity. In addition, the cytoprotective effects of c-FLIP L seem to be significantly stronger than those of c-FLIP S . Accordingly, in the present study, expression of c-FLIP L was induced in intact pig islets by adenoviral transduction. Consequently, the cytoprotective capacity of the transgene in pig islets was examined in in vitro and in vivo exposure to human CD8+ CTLs. Cells from untransduced islets or mock islets were sensitive to CD8+ CTL-mediated lysis (59.3% ± 15.9% and 64.0% ± 8.9% cytotoxicity, respectively). In contrast, cells from pig islets transduced with the c-FLIP L gene were markedly protected from lysis (30.5% ± 3.5%). Furthermore, prolonged xenograft survival was elicited from pig islets transduced with this molecule as assessed using an islet transplant model using the rat kidney capsule. Thus, these data indicate that intact pig islets can be transduced to express c-FLIP L with adenovirus. Pig islets expressing c-FLIP L are significantly resistant to human CTL killing and further exhibit beneficial effects to prolong xenograft survival.

Cannabinoids inhibit the respiration of human sperm

To investigate the effects of the psychotropic compounds Delta(9)-tetrahydrocannabinol (Delta(9)-THC) and Delta(8)-tetrahydrocannabinol (Delta(8)-THC) on sperm mitochondrial O(2) consumption (respiration). State University of New York Upstate Medical University, Syracuse, New York. Forty-one men who visited the andrology laboratory for fertility evaluation. None. A phosphorescence analyzer that measures O(2) concentration in sperm suspensions as a function of time was used to measure respiration. An immediate decline in the rate of respiration was observed when Delta(9)-THC or Delta(8)-THC was added to washed sperm. The inhibition was concentration dependent, and Delta(9)-THC was the more potent of the two compounds. Respiration was much less affected when Delta(9)-THC or Delta(8)-THC was added to neat semen, suggesting the presence of protective factors in seminal plasma. Both compounds inhibited the respiration of isolated mitochondria, illustrating that direct mitochondrial damage is likely the primary mechanism of action. The two main active cannabinoids of the marijuana plant, Delta(9)- and Delta(8)-THC, are potent inhibitors of mitochondrial O(2) consumption in human sperm. These findings emphasize the adverse effects of these toxins on male fertility. The cytoprotective capacity of seminal plasma deserves further investigation.