Herbal extracts counteract cisplatin-mediated cell death in rat testis

Aim： To evaluate the protective effects of ginger （Gin） and roselle （Ros） against testicular damage and oxidative stress in a cisplatin （CIS）-induced rodent model. Their protective effects against CIS-induced apoptosis in testicular and epididymal sperms is also investigated. Methods： Ethanol extracts of Gin or Ros （1 g/kg.day） were given orally to male albino rats for 26 days. This period began 21 days before a single CIS intraperitoneal injection （10 mg/kg body weight）. Results： Gin or Ros given orally significantly restored reproductive function. Both tested extracts notably reduced the CIS-induced reproductive toxicity, as evidenced by restoring the testis normal morphology. In Gin and Ros, the attenuation of CIS-induced damage was associated with less apoptotic cell death both in the testicular tissue and in the sperms. CIS-induced alterations of testicular lipid peroxidation were markedly improved by these plant extracts. Conclusion： The present results provide further insights into the mechanisms of protection against CIS-induced reproductive toxicity and confirm the essential anti-oxidant potential of both examined extracts. （Asian J Androl 2008 Mar; 10： 291-297）     

Sperm aggregation by water extracts from two Bursera species

Some plants have more than the common utility value, as is the case of some members of the Bursera species such as the Mexican copal, a plant used for worship. Water extracts of several plants have vaginal contraceptive properties. The authors evaluated the sperm agglutinating activity of two Bursera species on human and boar sperm. Extracts from stems and leaves were obtained. Capacitated sperm samples were used in all cases. There were different agglutinating capacities, which were not observed in the vehicle-only samples. The most frequent sperm agglutination response was that involving the heads. Agglutinating activity was higher from stem- than leaf-derived extracts. The results indicate that proteins present in the extracts are responsible for the aggregation of sperm heads.     

Population studies showing cross-reactivity of Toxoplasma gondii antibodies with antibodies to malignant cervical tissue antigens

Antibody inhibition studies have established that ethanol-soluble extracts prepared from a pool of malignant cervical tissues contained substances capable of reacting with antibodies specific for Toxoplasma gondii and malignant tissue factors. Extracts from normal cervical tissue did not possess Toxoplasma or malignant tissue antigens. Antibodies to malignant cervical tissue extract could be detected singly or in combination with Toxoplasma antibodies. The overall incidence of antibodies to malignant tissue extract in over 2,000 samples of blood tested was 4.4%. Of 98 apparently healthy donors with antibodies to malignant tissue extract, 50% also possessed Toxoplasma antibodies. The combined incidence of anti-Toxoplasma activity was 16.5% of all samples tested. Methods for the isolation and determination of IgG antibodies to malignant tissue and Toxoplasma antigen extracts are described, and the possible significance of the findings discussed.     

Saw palmetto extracts potently and noncompetitively inhibit human alpha1-adrenoceptors in vitro

BACKGROUND: We wanted to test whether phytotherapeutic agents used in the treatment of lower urinary tract symptoms have alpha1-adrenoceptor antagonistic properties in vitro. METHODS: Preparations of beta-sitosterol and extracts of stinging nettle, medicinal pumpkin, and saw palmetto were obtained from several pharmaceutical companies. They were tested for their ability to inhibit [3H]tamsulosin binding to human prostatic alpha1-adrenoceptors and [3H]prazosin binding to cloned human alpha1A- and alpha1B-adrenoceptors. Inhibition of phenylephrine-stimulated [3H]inositol phosphate formation by cloned receptors was also investigated. RESULTS: Up to the highest concentration which could be tested, preparations of beta-sitosterol, stinging nettle, and medicinal pumpkin were without consistent inhibitory effect in all assays. In contrast, all tested saw palmetto extracts inhibited radioligand binding to human alpha1-adrenoceptors and agonist-induced [3H]inositol phosphate formation. Saturation binding experiments in the presence of a single saw palmetto extract concentration indicated a noncompetitive antagonism. The relationship between active concentrations in vitro and recommended therapeutic doses for the saw palmetto extracts was slightly lower than that for several chemically defined alpha1-adrenoceptor antagonists. CONCLUSIONS: Saw palmetto extracts have alpha1-adrenoceptor-inhibitory properties. If bioavailability and other pharmacokinetic properties of these ingredients are similar to those of the chemically defined alpha1-adrenoceptor antagonists, alpha1-adrenoceptor antagonism might be involved in the therapeutic effects of these extracts in patients with lower urinary tract symptoms suggestive of benign prostatic obstruction.     

Influence of nociception and stress-induced antinociception on genetic variation in isoflurane anesthetic potency among mouse strains

BACKGROUND: Genetic background influences anesthetic potency to suppress motor response to noxious stimulation (minimum alveolar concentration [MAC]) as well as nociceptive sensitivity in unmedicated animals. However, the influence on MAC of baseline sensitivity to the noxious stimuli used to assess MAC has virtually never been studied. The authors assessed room air nociceptive sensitivity and isoflurane MAC in multiple mouse strains. Isoflurane requirement for loss of righting response (MACLORR) was also measured. METHODS: One outbred and 10 inbred mouse strains were tested for latency to respond (in room air) to a tail clip (either 500 g or 2,000 g). Naive mice of the same 11 strains were tested for isoflurane MAC and MACLORR. To assess the role of opioid-mediated stress-induced antinociception, mice were also tested for nociceptive sensitivity after injection of naloxone (10 mg/kg) or saline. RESULTS: Robust strain differences were observed for all measures. The authors found that tail-clip latency (using a 500-g or 2,000-g clip, respectively) correlated significantly with MAC (r = -0.76 and -0.58, respectively) but not MACLORR (r = -0.10 and -0.26). Naloxone produced strain-dependent reductions in open air tail-clip latencies, and these reductions were also strongly correlated with MAC (r = -0.67 and -0.71). CONCLUSIONS: The authors suggest that genetic variability in isoflurane MAC (but not MACLORR) may reflect genetic variability in the underlying sensitivity to the noxious stimulus being used to measure MAC. This variable sensitivity to nociception in the awake state is at least partially mediated by endogenous antinociceptive mechanisms activated by the tail-clip stimulus itself.     

Effect of extracts from the Chinese and European mole cricket on wound epithelialization and neovascularization: in vivo studies in the hairless mouse ear wound model

Until the end of World War II, oily extracts from the European mole cricket, Gryllotalpa gryllotalpa Linné , were used for treating nonhealing wounds and burns. In traditional Chinese medicine, extracts from the Chinese mole cricket, Gryllotalpa africana Beauvois , have been used to treat boils, abscesses, and ulcers successfully for over two centuries and are still being used today. The aim of this study was twofold: first, to measure the effect mole cricket extracts have on wound epithelialization and neovascularization, and second, to identify the active compounds in the Chinese and German mole cricket extracts. For the first aim, the hairless mouse ear wound model was used. The findings showed that wounds treated with the mole cricket extracts epithelialized significantly faster than control wounds 12.7±0.9 and 13.2±1.4 days vs. 16.3±2.2 days (mean±SD, p<0.05), respectively. While the rate of wound neovascularization was significantly increased in the first 3 days postwounding from that point on, the rate in treated wounds was the same as in controls. To identify the active compounds in the mole cricket extracts, the extracts were fractionated and tested in a foreskin basal keratinocyte cell culture assay. In this assay, the migration of keratinocytes is similar to skin cell migration or reepithelialization in a healing wound. Using this method, we found the active compound in the mole cricket extracts to be linoleic acid methyl ester. All other fatty acid structures that were isolated were found to be inactive.     

Oligoclonal immune response in cerebral cavernous malformations. Laboratory investigation

OBJECT: Mechanisms of cerebral cavernous malformation (CCM) pathogenesis include genetic predisposition in some cases, but other factors are likely to be involved in lesion proliferation and clinical manifestations. Given the unique antigenic milieu of CCMs, there may be a characteristic immune response in these lesions. We hypothesize that the immunoglobulin (Ig) fraction in CCMs reflects an oligoclonal immune response not present in paired sera from the same patients or in other types of cerebrovascular malformations. METHODS: Surgically excised lesions from five patients with CCMs, three patients with arteriovenous malformations (AVMs), and four normal brain control specimens obtained at autopsy were homogenized and extract tested for IgG clonality by isoelectric focusing in parallel with each patient's serum. RESULTS: The authors detected B cells in all three lesions examined, and plasmacytes in two out of three lesions examined. Four of five extracts of homogenized CCMs showed an oligoclonal pattern of IgG distinct from the polyclonal pattern seen in those patients' sera. Immununoglobulin G oligoclonality was not seen in AVMs or control brain specimens. CONCLUSIONS: The results of isoelectric focusing studies showed that CCM lesions had oligoclonal patterns of IgG unrelated to peripheral blood contamination, indicating selective synthesis of IgG within the lesions. This finding probably reflects a clonal expansion of B cells and/or plasmacytes in CCMs, an event that might be antigen-driven or a potential marker of inflammation.     

Improvement of penile erection, sperm count and seminal fructose levels in vivo and nitric oxide release in vitro by ayurvedic herbs

In the present study, the effect of four Vajikaran Rasayana herbs on penile erection, sperm count, seminal fructose content in vivo and nitric oxide (NO) release in vitro was assessed. Lyophilised aqueous extracts of Asparagus racemosus Willd. (AR), Chlorophytum borivilianum Sant. F. (CB), Curculigo orchioides Gaertn. (CO), and Dactylorhiza hatagirea (D. Don) Soo (DH) were orally administered at 100 mg/kg body weight to Wistar strain male albino rats. Penile erection index and sperm count were determined by visual observation; the seminal fructose concentration was measured spectrophotometrically using resorcinol reagent; and NO release was assessed in a mouse macrophage cell line (RAW264) spectrophotometrically using a commercial Griess reagent kit. Penile erection index, sperm count, seminal fructose concentration and in vitro NO release were the parameters measured. A significant effect on the sperm count, seminal fructose content and penile erection index was observed upon treatment with the extracts. The effect of extracts on inducible NO release in vitro directly correlated with the enhanced erectile function in vivo. The aphrodisiac claims attributed to the four Vajikaran Rasayana herbs were tested and a distinctive effect of all extracts tested was observed, with C. borivilianum showing a highly significant response for all parameters measured in vivo and in vitro. The present study also provides a good correlation between the in vivo improvement of penile erection and in vitro NO releasing activity of the extracts. Increase in seminal fructose levels and sperm count further validates the role of these herbs in improving reproductive function.     

Metastatic potential of colon carcinoma. Expression of ABO/Lewis-related antigens

Four monoclonal antibodies specific to various Lewisx-related antigens were tested for binding to monolayers and detergent extracts of the human colon carcinoma cell line HT-29 P and its metastatic variant HT-29 LMM. Only monoclonal antibody FH6 (antisialyl-dimeric Lex) bound more to the metastatic variant compared with HT-29 P. Detergent extracts of human tissues were then assayed for their FH6-binding activity. Normal colonic mucosa displayed less FH6 binding than either Dukes stage B or D primary colon carcinomas. Liver metastases from colorectal carcinoma demonstrated greater binding than stage B and stage D primary tumors, and more than normal liver. Expression of sialyl-dimeric Lex increases as the metastatic potential of colorectal carcinomas increases, and is related to the progression of colorectal cancer.     

Influence of Nociception and Stress-induced Antinociception on Genetic Variation in Isoflurane Anesthetic Potency among Mouse Strains

Background: Genetic background influences anesthetic potency to suppress motor response to noxious stimulation (minimum alveolar concentration [MAC]) as well as nociceptive sensitivity in unmedicated animals. However, the influence on MAC of baseline sensitivity to the noxious stimuli used to assess MAC has virtually never been studied. The authors assessed room air nociceptive sensitivity and isoflurane MAC in multiple mouse strains. Isoflurane requirement for loss of righting response (MACLORR) was also measured.

Methods: One outbred and 10 inbred mouse strains were tested for latency to respond (in room air) to a tail clip (either 500 g or 2,000 g). Naive mice of the same 11 strains were tested for isoflurane MAC and MACLORR. To assess the role of opioid-mediated stress-induced antinociception, mice were also tested for nociceptive sensitivity after injection of naloxone (10 mg/kg) or saline.

Results: Robust strain differences were observed for all measures. The authors found that tail-clip latency (using a 500-g or 2,000-g clip, respectively) correlated significantly with MAC (r = -0.76 and -0.58, respectively) but not MACLORR (r = -0.10 and -0.26). Naloxone produced strain-dependent reductions in open air tail-clip latencies, and these reductions were also strongly correlated with MAC (r = -0.67 and -0.71).     

Experimental cryptorchidism inhibited growth of the rat ventral prostate

Adult Sprague-Dawley male rats, weighing about 350 g, were rendered cryptorchid by suturing the testes to the lateral abdominal wall. Twenty-eight days later, cryptorchidism resulted in a significant decline in testis weight and suppressed spermatogenesis. The ventral prostate was significantly smaller in cryptorchid rats. There was no significant difference in serum testosterone levels between the normal and cryptorchid rats. Charcoal-stripped aqueous extracts of the testis from intact and cryptorchid animals were tested on primary cultures of rat prostatic stromal cells. Cultures treated with extract from the intact testis had a significantly increased cell proliferation as assessed by cell count and by the rate of 3H-thymidine incorporation. Additionally, extracts of seminiferous tubules significantly increased prostate stromal cell proliferation compared to extracts of testicular interstitial components. Furthermore, this proliferative effect of testicular extracts is specific to the prostate as extract of both normal and cryptorchid testis stimulated proliferation of rat footsole fibroblasts in culture, but only extracts from intact testis stimulated proliferation of prostate stromal cells. These observations demonstrate that the testis produces nonandrogenic substances that can promote growth of prostatic stromal cells and that these substances were eliminated in the cryptorchid testis.     

Reactivity of gastric cancer patients in leucocyte migration inhibition tests to 3M KCl extracts from gastric tumor

Leucocytes from patients with gastric cancer and other malignant and non-malignant diseases of the gastrointestinal tract as well as from healthy controls were tested for leucocyte migration inhibition test (LMI) using five different, allogeneic 3M KCl soluble extracts from gastric cancer tissues. The normal range of migration index (MI) was considered to be between. 0.77 and 1.18 by calculating the mean MI±2SD of ten healthy controls with cancer extracts. MIs out of this range were considered to be pathologic. In LMI test with a single tumor extract, pathologic MI was found in 48% of 79 gastric cancer patients, such being significantly higher than in those (4–21%) of three other groups of patients. In the panel model of LMI, i.e., testing each blood sample with five different tumor extracts, 79% (62/79) of patients with gastric cancer were reactive, while 25% (5/20) of colorectal cancer patients showed “positive” reaction and no “positive” reactivity was observed in two other groups of patients. thus, the “positive” reactivity in patients with gastric cancer was observed significantly higher than those in the other 3 groups of patients. Gastric cancer extracts had a wide range of cross-reactivity when compared with colorectal tumor extracts which showed a relatively restricted corss-reactivity. Thus, the LMI test, particularly when tested by a panel mode, seems to express cell-mediated immunity against tumor associated antigens of gastric cancer.     

Antispermatogenic and hormonal effects of Crotalaria juncea Linn. seed extracts in male mice

Aim: To evaluate the antifertility activity of various extracts of Crotalaria juncea seeds in male mice. Methods: Adult male mice were gavaged the petroleum ether, benzene and ethanol extracts of C. juncea seeds, 25 mg·(100g)-1·day-1 for 30 days. On day 31 the animals were sacrificed by cervical dislocation and the testes, epididymis, vas deferens, seminal vesicles, prostate gland, bulbourethral gland and levator ani were dissected out and weighed. The organs were processed for biochemical and histological examination. Results: In petroleum ether, benzene and ethanol extracts treated rats, there was a decrease in the weights of testis and accessory reproductive organs. The diameters of the testis and seminiferous tubules were decreased. Spermatogonia, spermatocytes and spermatids in the testis and the sperm count in cauda epididymis were also decreased. There was a significant reduction in the protein and glycogen contents and an increase in the cholesterol content in the testis, epididymis and vas def     

In vitro measurement of antibacterial activities of topical antibacterial creams]

A new method of in vitro measurement of antibacterial activity of different antimicrobial creams is introduced in this paper. The tested cream in known quantity is uniformed suspended in a solid medium, and bacteria of known quantity are inoculated. The result is expressed in MIC. In this observation, four creams were tested against 3 bacteria. It is the authors impression that the new method is better than the traditional disc method, as it reflects better the actual antibacterial activity of the drug in a cream base. It is helpful in selecting an effective drug or to evaluate the effectiveness of certain drugs in clinical practice.     

Autosensitization of neurosurgical cases by normal brain tissue antigens (author's transl)]

The leukocyte reactions of 106 neurosurgical cases, including 63 brain tumors, 10 intracerebral hematomas, 10 cerebral infarctions, 10 subarachnoidal hemorrhages, 8 cerebral injuries and 5 chronic subdural hematomas, against the extracts of gliomas and normal brain tissues were tested by capillary migration (LMI) and adherence inhibition (LAI) assays. Both tests showed specific responses with autochthonous and allogeneic glioma extracts in glioma patients. The sensitivity of LAI was superior to that of LMI, although LAI also showed adherence enhancement in the presence of weakly sensitized leukocytes or weak antigenic stimuli. Leukocytes from glioma patients showed positive inhibition with normal brain tissues from patients with glioma and intracerebral hematoma. Positive leukocyte reactions with normal brain tissues were also confirmed in patients with intracerebral hematomas, cerebral infarctions and severe cerebral lacerations, but not in those with subarachnoidal hemorrhages, minor cerebral contusions and chronic subdural hematomas. These results suggest that the leukocytes of patients with destructive brain lesions were autosensitized by normal brain antigens. The autosensitization has some advantages in that destroyed brain tissues are eliminated, but the hyperimmune state might cause postictal brain edema and should be properly controlled by steroids.     

Hollow-Fiber Dialyzers and Their Pyrogenicity Testing by Limulus Amebocyte Lysate

Aqueous extracts of cellulose hollow fibers (CHF) exhibit positive reactions in some Limulus amebocyte lysate (LAL) tests. However, in spite of LAL activity, the extracts produce no fever reaction in rabbits. A comparison of lysates from different suppliers shows pronounced activity differences when extracts of cuprammonium-derived CHF are tested. One of the lysates, which is fully reactive against standard endotoxin, shows no reaction with such extracts, nor do CHF extracts diminish its sensitivity to standard endotoxin. Investigations of the cuprammonium process have shown that endotoxins introduced by the linters are degraded and washed out. Other endotoxin introduction, particularly by the process water, has been excluded. Oxidative or acidic degradation of cellulose does not result in the formation of LAL-reactive material (LAL-RM). On the other hand, sterile cotton wool shows LAL reactivity, and cellulose acetate regains LAL reactivity when it is saponified. Thus, it appears likely that the LAL-RM found in CHF is of purely cellulosic origin and crossreacts with a number of commercially available lysates.     

Prevalence of positive preoperative pregnancy testing in teenagers scheduled for orthopedic surgery

The pregnancy status of female teenagers presenting for orthopaedic surgery is often unknown. Anesthetics may have teratogenic or abortive effects. The authors report the results of a screening program for unrecognized pregnancy in 532 female teenagers presenting for orthopaedic surgery. Five patients with a positive urine human chorionic gonadotropin test were identified, for a prevalence of 1 positive result in 106 patients tested (0.9%). Surgery was cancelled in each case.     

Natural killer cells in patients with renal cell cancer

Cytotoxic activity of natural killer (NK) cells in peripheral blood was studied in 60 patients with renal cell carcinoma (RCC) by means of a 51Cr release assay against K 562 cells, homologous and autologous RCC. All patients were tested prior to surgery and thereafter followed up for 1 year. Immunostimulating agents as interferon, levamisole, OMPI, and various bacterial extracts were tested for their boosting capacity on NK cells. Patients with localized tumor showed increased activity in comparison to age-matched controls, whereas NK cell activity and interferon-boosted activity in patients with advanced disease and tumor spread was decreased. Augmentation of NK activity could only be achieved by interferon, while application of levamisole, OMPI, and bacterial extracts resulted rather in suppression of cytotoxic activity.     

Finding orthopedic patients lost to follow-up for long-term outcomes research using the Internet: an update for 2012

The Internet is available to researchers as a tool for studying long-term outcomes, but no recent research exists on how to best use it. The authors hypothesize that using the Internet can be at least 75% effective in locating patients lost to follow-up. With Institutional Review Board approval, the authors searched for 66 patients lost to follow-up after a period of 10 years or more with no contact. They tested an Internet searching protocol developed in 2004 and developed an alternate protocol. In all, 74% (49/66) of patients were located.     

Activation of latent transforming growth factor beta1 by stromelysin 1 in extracts of growth plate chondrocyte-derived matrix vesicles.

Previous studies have shown that matrix vesicles isolated from cultures of costochondral growth zone chondrocytes and treated with 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] can activate recombinant human latent transforming growth factor beta1 (rhTGF-beta1). It is unknown what enzyme or other factor in the extracellular organelles is responsible for the activation. This study tested the hypothesis that enzymes present in matrix vesicles can activate latent TGF-beta1 and that this is regulated by 1alpha,25(OH)2D3. To do this, we examined the ability of matrix vesicle extracts to activate small latent rhTGF-beta1. In addition, enzymes previously determined to be present in matrix vesicles were screened for their ability to activate small latent rhTGF-beta1. Recombinant human matrix metalloproteinase 2 (rhMMP-2; 72 kDa gelatinase), rhMMP-3 (stromelysin 1), purified human plasminogen, and purified urokinase (plasminogen activator) were each tested at varying concentrations. To assess the role of cell maturation, we used a cell culture model in which chondrocytes are derived from two distinct zones of rat costochondral cartilage, the resting zone and the growth zone. Matrix vesicles were isolated from these cultures and then tested. The results showed that extracts of matrix vesicles produced by both growth zone and resting zone chondrocytes were able to activate small latent rhTGF-beta1. The effects were dose and time dependent, with greater activity being found in extracts of matrix vesicles from the growth zone chondrocyte cultures. Only rhMMP-3 was able to activate small latent rhTGF-beta1, indicating that stromelysin-1, but not MMP-2, plasminogen, or urokinase, was involved. As observed in the extracts, the effect of rhMMP-3 was time and dose dependent. When anti-MMP-3 antibody was added to matrix vesicle extracts from both cell types, activation of small latent rhTGF-beta1 was dose-dependently blocked. Neither 1alpha,25(OH)2D3 nor 24R,25(OH)2D3 had a direct effect on activation of small latent rhTGF-beta1 by the extracts. However, when intact matrix vesicles were treated with 1alpha,25(OH)2D3, their ability to activate small latent rhTGF-beta1 was increased. Inhibition of phospholipase A2 with quinacrine blocked the 1alpha,25(OH)2D3-dependent effect. These results suggest that the ability of 1alpha,25(OH)2D3-treated matrix vesicles to activate small latent TGF-beta1 is via action of the secosteroid on the matrix vesicle membrane, not on the enzymes responsible for activating latent TGF-beta1. Because matrix vesicles isolated from growth zone chondrocytes have been shown to contain increased phospholipase A2 activity after treatment with 1alpha,25(OH)2D3, it is likely that this secosteroid promotes loss of membrane integrity through phospholipase A2-dependent formation of lysophospholipids, resulting in the release of MMP-3 into the matrix, where latent TGF-beta1 is stored. Taken together, the results of the current study show that matrix vesicles produced by growth plate chondrocytes contain MMP-3, that this enzyme is at least partially responsible for activation of small latent TGF-beta1 in the matrix, and that 1alpha,25(OH)2D3 regulates MMP release from matrix vesicles.