[Cannabidiol: its use in refractory epilepsies]. / Cannabidiol: uso en epilepsias refractarias.

INTRODUCTION: Some epileptic syndromes are characterised by seizures that are difficult to control and are associated to delayed neuropsychomotor development, which results in a deterioration in the patient's quality of life as well as in that of his or her family. AIM: To evaluate the use of cannabidiol as adjuvant therapy in patients with refractory epilepsies. PATIENTS AND METHODS: An observational study was conducted by means of a survey addressed to the patient's caregiver. Data collected included information about the patient and the caregiver, changes observed in the seizures, neuropsychological effects, side effects and the family's overall perception following the use of cannabidiol. RESULTS: The evaluation examined 15 patients with refractory epilepsies, who received cannabidiol over a period ranging from one month to one year. The frequency of seizures decreased in 40% of the patients, 60% of the patients were seen to have control over 50% of their seizures and in 27% of them the seizures disappeared completely. Neurocognitive changes were also reported: behaviour improved in 73%; 60% reported an improvement in language; in 50% sleep improved; 43% reported improvements in eating habits; and 100% said their mood had improved. The overall perception of the illness was that there had been improvements in 73% of respondents. The most common side effects were drowsiness and fatigue. CONCLUSIONS: These results suggest a possible beneficial effect of cannabidiol on the control of seizures and on the improvement of certain neurocognitive aspects in patients with refractory epilepsies.

TITLE: Cannabidiol: uso en epilepsias refractarias.Introduccion. Algunos sindromes epilepticos se caracterizan por crisis de dificil control y asocian un retraso en el desarrollo neuropsicomotor, lo que conlleva un deterioro en la calidad de vida del paciente y su familia. Objetivo. Evaluar el uso del cannabidiol como tratamiento adyuvante en pacientes con epilepsias refractarias. Pacientes y metodos. Se realizo un estudio observacional por medio de una encuesta dirigida a la persona cuidadora del paciente. Se valoro la informacion sobre el paciente y el cuidador, cambios observados sobre las crisis, efectos neuropsicologicos, efectos adversos y percepcion global de la familia tras el uso del cannabidiol. Resultados. Se evaluo a 15 pacientes con epilepsias refractarias, quienes recibieron cannabidiol durante un periodo de un mes a un año. En el 40% de los pacientes hubo una disminucion en la frecuencia de las crisis, en el 60% de los pacientes se observo un control de mas del 50% de las crisis y en el 27% las crisis desaparecieron totalmente. Tambien se comunicaron cambios neurocognitivos: en el 73% hubo una mejoria del comportamiento; el 60% notifico una mejoria en el lenguaje; el 50%, en el sueño; el 43%, en la alimentacion; y el 100%, en el estado de animo. La percepcion global sobre la enfermedad notifico una mejoria en el 73%. Los efectos adversos mas frecuentes fueron somnolencia y fatiga. Conclusiones. Estos resultados sugieren un posible efecto beneficioso del cannabidiol sobre el control de las crisis y en la mejoria de ciertos aspectos neurocognitivos en pacientes con epilepsias refractarias.

Functionalized magnetic nanoparticles as vehicles for the delivery of the antitumor drug gemcitabine to tumor cells. Physicochemical in vitro evaluation.

Gemcitabine is a chemotherapy drug used in different carcinomas, although because it displays a short biological half-life, its plasmatic levels can quickly drop below the effective threshold. Nanoparticle-based drug delivery systems can provide an alternative approach for regulating the bioavailability of this and most other anticancer drugs. In this work we describe a new model of composite nanoparticles consisting of a core of magnetite nanoparticles, coated with successive layers of high molecular weight poly(acrylic acid) and chitosan, and a final layer of folic acid. The possibility of using these self-assembled nanostructures for gemcitabine vehiculization is explored. First, the surface charge of the composite particles is studied by means of electrophoretic mobility measurements as a function of pH for poly(acrylic acid) (carbopol) of different molecular weights. The adsorption of folic acid, aimed at increasing the chances of the particles to pass the cell membrane, is followed up by optical absorbance measurements, which were also employed for drug adsorption determinations. As a main result, it is shown that gemcitabine adsorbs onto the surface of chitosan/carbopol-coated magnetite nanoparticles. In vitro experiments show that the functionalized magnetic nanoparticles are able to deliver the drug to the nuclei of liver, colon and breast tumor cells.

Comparison between open and laparoscopic appendectomy in non-complicated appendicitis

BACKGROUND/OBJECTIVE: An extensive web search failed to provide studies from Puerto Rico regarding whether open (OA) or laparoscopic appendectomy (LA) should be performed for non-complicated appendicitis. Our goal is to compare these techniques in terms of time at operating room (OR), length of surgery, hospital stay, pain medication requirements, in-hospital complications and readmissions. METHODS: 126 patients (64 OA; 62 LA) with non-complicated appendicitis were studied retrospectively. Data obtained: demographics, CT-Scan use, surgery and operating room time, days in hospital, complications, diet commencement, pain medications doses, pathology and readmission. RESULTS: Difference was found in total time at OR (80.1+/-29 minutes OA; 105.7+/-22.6 LA) and in surgery length (41+/-28 OA; 48+/-16 LA), but not in hospital stay (2.1 days OA; 2.2 LA) nor in in-hospital complication rate. Negative appendectomy rate was 24% LA vs. 3% OA. Readmission rate was higher in OA with 5% wound infection rate. CONCLUSION: Techniques are similar in mean hospital stay, in-hospital complications, and pain medication requirements. LA had a higher negative appendectomy rate but of these patients five had surgical diagnosis of acute appendicitis and after appendectomy, signs and symptoms resolved; and two patients had interval appendectomies. As these patients were cured, the real negative appendectomy rate is 13%, similar to the historically accepted 16%. The other eight patients had an adequate diagnosis. We are concerned OA negative appendectomy rate is only 3%; we wonder if surgeons are waiting too long to operate patients. Readmission was higher in OA (wound infection rate of 5%). Although it takes more time in the OR, LA is as safe as OA, has a low rate of complications and lower readmission rate.

Experience with hand assisted laparoscopic surgery of the colon

BACKGROUND: Hand assisted laparoscopic colectomy (HALS) has been shown to have the advantages of laparoscopic colectomy in terms of pain, recovery and length of hospital stay. Studies have shown similar outcomes in laparoscopic colectomy as in open surgery. There is a learning curve to HALS, the operative time is longer, and it is more difficult than open surgery and requires specialized equipment. In this report we present our initial experience over a 2.5 year period using HALS for colon surgery for diverticulosis, polyps and colon cancer. METHODS: A retrospective review of office and hospital charts of patients undergoing HALS colectomy from June 2005 to January 2008 was performed at HIMA-San Pablo Hospital. Demographics, outcomes data including operative time, conversion rate to open surgery, reasons for conversion, time to start feedings, and length of stay were collected as well as staging and number of nodes for cancer patients. Complications are discussed along with comments pertinent to the experience of two surgeons going through the learning curves of LC and HALS colectomy. RESULTS: A total of 65 patients underwent attempted hand assisted laparoscopic colon resection. There were 33 males and 32 females between the ages of 26 and 87. Thirty-one patients underwent surgery for diverticulosis; 8 for pre-malignant lesions (large polyps or polyps with high grade dysplasia), and 26 for colon cancer. Mean operative time was 195 minutes (120 to 300); mean length of stay was six days (range 4-14 days). Conversion rate was (13.8%) overall; 21% during the first year and 10.8% after the first year. 5 (7.5%) of the patients in which HALS colon resection was completed had complications with prolonged length of stay. Patients without complication had an average length of stay of 4.5 days. The average number of lymph nodes was 14.8 (range 7-24); average length of specimens for diverticulosis was 17cm. Complications included postoperative bleeding in three patients...

Effects of intermittent exposures of aflatoxin B(1) on hepatic and testicular glutathione S-transferase in rats.

Glutathione S-transferase (GST) plays a major role in the detoxification of the potent hepatocarcinogen aflatoxin B(1) (AFB(1)). This study evaluated the effects of intermittent exposures to AFB(1) on hepatic and testicular GST in rats. Male Fischer 344 rats were fed diets containing AFB(1) (0, 0.01, 0.04, 0.4 and 1.6 ppm) intermittently at 4-week intervals up to 20 weeks. The control animals were fed an AFB(1)-free NIH-31 diet. Rats consuming diets with 0.01 ppm AFB(1) did not show the induction of hepatic or testicular GST activity. Intermittent exposures to AFB(1) at concentrations of 0.04-1.6 ppm significantly increased the GST activities. The increase of the enzyme activity was proportional to the dose and length of AFB(1) exposure.

Unscheduled DNA synthesis assay in mammalian spermatogenic cells: an update.

The unscheduled DNA synthesis (UDS) assay measures DNA repair in response to DNA damage. To date, 59 chemicals plus UV and X rays have been tested for UDS in spermatogenic cells of humans, rabbits, rats, and mice. In vivo, in vitro, and combined in vivo/in vitro procedures have been used. UDS has been shown to occur in spermatogonia, meiotic spermatocytes, and early spermatid stages. Fifty-nine percent of the agents tested gave a positive UDS response in one or more germ-cell stages. Results show 95% concordance (positive or negative) between different mammalian species. Some well-known genotoxic chemicals, for example, aflatoxin B(1) (AFB(1)), benzo[a]pyrene (B[a]P), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), did not induce significant levels of UDS. Possible explanations are discussed. Results from the UDS assay were compared with those from the mouse specific-locus mutation (SLM) test to determine correlations between the two assays. Only two chemicals, ethyl- and methyl-nitrosourea (ENU and MNU), have been tested for UDS and SLM induction in spermatogonial stages. Results show full concordance between the two assays. In postspermatogonial stages, 25 chemicals and X rays have been tested for UDS and SLM induction. Seventy-seven percent of these agents showed similar results (positive or negative) in these germ-cell stages. Although the UDS assay cannot replace the SLM test, the strong correlations between the two assays suggest the usefulness of the UDS assay as a predictor of germ-cell mutations in mammalian systems.

Differential sensitivities of spermatogonial and postspermatogonial cell stages of the mouse to induction of unscheduled DNA synthesis by ethyl- and methyl-nitrosourea.

An autoradiographic procedure was used to measure unscheduled DNA synthesis (UDS, DNA repair synthesis) in spermatogonial and postspermatogonial cell stages of mice after treatment with two doses of N-ethyl-N-nitrosourea (ENU) and N-methyl-N-nitrosourea (MNU). Significant levels of UDS were measured in type A spermatogonia, meiotic spermatocytes, round spermatids, and early elongating spermatids but not in mature spermatids. The extent of UDS varied according to the germ cell stage and the dose. At equimolar concentrations, MNU was more efficient than ENU in eliciting a UDS response in all germ cells. After ENU treatment, type A spermatogonia showed the highest UDS response, while round and elongating spermatids showed the lowest. After MNU treatment, pachytene spermatocytes exhibited the highest UDS response while type A spermatogonia showed the lowest. The high UDS response of type A spermatogonia to ENU parallels the well-known high mutational sensitivity of spermatogonia to this chemical. Similarly, the high UDS response observed in meiotic spermatocytes and early spermatid stages after MNU treatment correlates with the high mutational sensitivity of postspermatogonial stages to MNU. Thus, the present results, like the specific locus mutation studies, indicate that ENU and MNU each has a unique effect on the spermatogenic cells. This effect is likely due to the different mechanism of action of ENU and MNU at the level of DNA and also to the physiological differences between different germ-cell stages. Teratogenesis Carcinog. Mutagen. 19:339-351, 1999. Published 1999 Wiley-Liss, Inc.

Temporal patterns of DNA adduct formation and glutathione S-transferase activity in the testes of rats fed aflatoxin B1: a comparison with patterns in the liver.

Fisher-344 male rats were fed 1.6 ppm of aflatoxin B1 (AFB1) continuously and intermittently for several weeks. At various time periods, DNA was isolated from the testes and livers and analyzed for AFB1-DNA adducts. The ability of the testis to detoxify AFB1 was also investigated by the glutathione S-transferase (GST) activity assay and compared with that of the liver. The levels of testicular AFB1-DNA adducts were 2.4 to 8.1 times lower than those of the liver after 4 to 16 weeks of continuous treatment and 2.2 to 46.2 times lower after 8 to 20 weeks of intermittent treatment. The testicular DNA adducts markedly decreased over time. By 16 weeks of continuous and 20 weeks of intermittent exposure, they had decreased 37 and 91%, respectively. In contrast, hepatic AFB1-DNA adducts increased four-fold from 4 to 16 weeks of continuous treatment but increased at a much slower rate after intermittent exposure. In both the liver and testis, significant levels of AFB1-DNA adducts persisted for at least 1 month after ending the treatment, suggesting that this type of lesion was poorly repaired. In control rats, the testis showed significantly higher GST activity than the liver. In treated rats, these differences were significant during the first 12 weeks of continuous treatment but not at later times. Tissue-specific differences such as germ-cell depletion and increased testicular detoxification may play an important role in the observed differential pattern of DNA adduct formation between the testis and liver.

A procedure to study unscheduled DNA synthesis in rat spermatogenic cells in vivo: strain differences between Fisher-344 and Sprague-Dawley rats.

A liquid scintillation counting (LSC) procedure was used to study unscheduled DNA synthesis (UDS) in pachytene spermatocytes and spermatids from Fisher-344 (F344) and Sprague-Dawley (SPD) rats treated with methyl methanesulfonate (MMS). MMS induced a large, dose-dependent, UDS response in pachytene spermatocytes from both rat strains. On average, F344 pachytene spermatocytes showed a larger UDS response than those of SPD rats. The lowest dose of MMS that elicited a significant UDS response was 1 mg/kg in F344 rats but 5 mg/kg in SPD rats. Early spermatid stages from F344 rats also showed a larger UDS response than those from SPD rats. The time interval at which spermatid stages showed the maximum UDS response was between 20 and 24 days after MMS treatment. It is concluded that UDS can be measured quantitatively in rat spermatogenic cells in vivo by using the LSC procedure.

Cryosurgery on the Treatment of Hepatic Tumors.

BACKGROUND: Primary and metastatic tumors in the liver are difficult to treat. When surgical resection is not feasible, cryotherapy is one of the several alternative approaches. METHODS: The data on outcomes from hepatic resections are reviewed, and the rationale and techniques of performing cryosurgery for unresectable hepatic cancers are described. The literature is reviewed and combined with the experiences of the authors on cryosurgery for management of hepatic tumors. RESULTS: The indications and techniques for performing cryosurgery are now well established. The procedure is relatively safe, and long-term survival rates of over 20% may be achieved. CONCLUSIONS: While cryotherapy is effective for localized tumors in the liver, additional adjuvant approaches are required to control disease in the untreated liver. Endoscopic techniques may minimize patient morbidity.

Formation of etheno and oxoethyl adducts in liver DNA from rats exposed subchronically to urethane in drinking water and ethanol.

Exposure of Fisher-344 male rats to 10 000 ppm of urethane in drinking water for up to 90 days or in 5% ethanol for up to 14 days caused the formation of 7-[2'-oxoethyl]guanine (OEG) and 1,N(6)-ethenoadenine (epsilon A) in liver DNA. Mild-acid DNA hydrolysates were analyzed by high-performance liquid chromatography with photodiode array detection and fluorometry. The identification of OEG and epsilon A was confirmed by coelution with the authentic standards. Forty and 67% of rats showed OEG and epsilon A adducts at 2 and 90 days of treatment with urethane in drinking water, respectively. In comparison, only 0 and 10% of rats showed adducts at 2 and 14 days of treatment with urethane in 5% ethanol, respectively. Neither OEG nor epsilon A was observed in control rats receiving water or 5% ethanol. Although these data are still preliminary, they appear to suggest that ethanol may inhibit formation of DNA adducts by urethane. Studies designed to produce more conclusive information about the role of ethanol in modifying DNA damage induced by urethane in vivo are in progress.

Unusual cases of hyperparathyroidism.

The history, physical and radiologic findings, treatment and pathology in five unusual cases of hyperparathyroidism is presented. The hyperparathyroidism was caused by a large (113 grams) mediastinal adenoma in the first patient, who is alive 25 years after surgery. A parathyroid carcinoma with compression of the esophagus was documented in the second patient. This patient is alive and normocalcemic 23 years after surgical treatment. A third patient with hyperplasia returned with hypercalcemia 20 years postsurgery requiring reoperation. A fourth patient with advanced bone findings was found to have a parathyroid adenoma. The fifth case is a patient with tertiary hyperparathyroidism secondary to hypophosphatemic rickets.

Induction of DNA damage by urethane in mouse testes: DNA binding and unscheduled DNA synthesis.

The extent and persistence of DNA damage and repair were investigated in mouse spermatogenic cells exposed in vivo to urethane (ethyl carbamate, EC). Adult male mice exposed to [3H]EC at 10-1,000 mg/kg were sacrificed 12 hr later. EC/metabolite binding to liver and testicular DNA and to sperm heads from the vasa deferentia was measured. Other male mice were exposed to EC at 50-750 mg/kg, and unscheduled DNA synthesis (UDS) induction was investigated in early spermatid stages. Similar experiments were conducted with vinyl carbamate (VC; putative EC metabolite) at 10-75 mg/kg. [3H]EC bound to liver and testicular DNA and to whole sperm heads. Testicular DNA binding increased linearly with dose, although binding was at least 2 orders of magnitude lower than with liver DNA. Sperm head binding also increased linearly with dose. Dose response studies with the UDS assay showed that EC and VC induced a small but significant increase of the UDS response in early spermatid stages. However, the induced UDS responses were quite variable and did not consistently increase with the administered dose. To determine the time kinetics of UDS induction, [3H]dThd was injected at various times after treatment with 500 mg/kg of EC or 60 mg/kg of VC. A slight but significant UDS increase was observed 4 hr after treatment with EC but not with VC. Overall, these results suggest that EC metabolites bind to testis DNA and cause low-level DNA damage in mouse spermatogenic cells. This type of DNA damage apparently does not have significant genetic consequences.

Mutagenicity, metabolism, and DNA interactions of urethane.

Urethane, a known animal carcinogen, has been the subject of intensive research efforts spanning 40 years. Recent concerns have focused on the presence of urethane in a variety of fermented foods and alcoholic beverages, although no epidemiological studies or human case reports have been published. Much information is available about the mutagenesis, metabolism, and DNA interactions of urethane in experimental systems. Urethane is generally not mutagenic in bacteria although in some instances it acts as a weak mutagen. Urethane is not mutagenic in Nuerospora but is weakly mutagenic in Saccharomyces. Drosophila appear to be the only organisms that consistently give positive mutagenic results with urethane, but its mutagenicity is weak and in many cases shows no clear dose dependence. Urethane is a good clastogen in mammalian somatic cells in vivo, but it shows variable results with cells in vitro. It efficiently induces sister chromatid exchanges in a variety of cells. Mammalian spermatogenic cells are insensitive to the induction of specific locus and dominant lethal mutations by urethane. Mutational synergism has been reported to occur between ethyl methanesulfonate and urethane when administered two generations apart, and some investigators have suggested possible synergism for cancer-causing mutations in mice exposed to X-rays and urethane one generation apart. These studies are controversial and have not been confirmed. Studies on the induction of cancer-causing dominant mutations by urethane are at variance with results from extensive studies with the specific locus test in mice. Urethane studies with the unscheduled DNA synthesis assay in mouse spermatogenic cells and with the sperm abnormality test have given negative results. Urethane is rapidly and evenly distributed in the body. The rate of elimination of urethane from plasma is a saturable process and varies according to the strain and age of the animal. Recent studies have concentrations similar to those in wine, ethanol inhibits the tissue distribution of urethane in mice. These results are important because they suggest a lower carcinogenic/mutagenic risk than expected from exposure to urethane in alcoholic beverages. Although research on the metabolic activation of urethane has been extensive, no conclusive results have been obtained about its active metabolite, at one time thought to be N-hydroxyurethane. More recently, it has been postulated that urethane is activated to vinyl carbamate and that this metabolite is capable of reacting with DNA.(ABSTRACT TRUNCATED AT 400 WORDS)

Failure of acrosome assembly in a male sterile mouse mutant.

Blind-sterile (bs) is a new autosomal recessive mutation of the mouse that causes sterility in males and bilenticular cataracts in both sexes. Sterile bs/bs males exhibited normal copulatory behavior, reduced testis weights, and few or no epididymal sperm. The effects of the bs mutation on spermatogenesis were examined by light and electron microscopy. All sperm present were morphologically abnormal with aberrant head shape. Adult bs/bs testes were characterized by germ cell depletion that resulted in profound alterations of the typical germ cell associations. Only 30% of the tubules contained relatively normal germ cell associations while 39% were extensively depleted, showing only Sertoli cells or Sertoli cells and spermatogonia. The most striking effect of the bs mutation on spermiogenesis was the failure of acrosome formation. Disorganized proacrosomic granules were detected up to step 3 of spermiogenesis by both periodic acid-Schiff staining and ultrastructural analysis. In over 3500 spermatids scored past steps 3-4 of spermiogenesis not a single acrosomal cap or fully developed acrosome was detected. Electron microscopy revealed a thickening of the nuclear envelope of elongating spermatids in the region where the acrosome should have been located; however, no acrosome was present. Chromatin condensation and nuclear elongation did occur in these acrosomeless spermatids, suggesting that caudal growth of the acrosome is not a mechanistic factor in these events.

Induction of unscheduled DNA synthesis in the germ cells of male mice after treatment with hydrazine or procarbazine.

The induction of unscheduled DNA synthesis (UDS) in early spermatid stages of mice was studied after treatment with hydrazine or procarbazine (Natulan). The UDS response was assayed by measuring the levels of [3H]dThd in the nuclei of sperm cells derived from treated and untreated early spermatid stages. The radioactivity present in these nuclei was measured by liquid scintillation counting. Mice were also treated with 100 mg/kg of MMS in order to compare the UDS level induced by this chemical with that induced by hydrazine or procarbazine. The level of UDS induced by hydrazine was not significantly different from that of the untreated controls in any of the 5 doses tested (10-120 mg/kg). Procarbazine, on the contrary, induced UDS in all the 5 doses tested, from 50 to 600 mg/kg. The increase in the UDS response was positively correlated with the increasing dose. The present results with hydrazine indicate either that this compound does not reach the germ cells in sufficient amounts to produce DNA damage or that the repair system in early spermatid stages does not recognize the type of DNA damage produced by this compound. The sensitivity of detection of UDS in the germ cells of male mice is compared with that of the rabbit. At equivalent germ-cell stages and dosage the mouse UDS assay is more sensitive, by a factor of 2 to 3, than the rabbit assay for the induction of UDS by procarbazine and MMS.

An autoradiographic study of unscheduled DNA synthesis in the germ cells of male mice treated with X-rays and methyl methanesulfonate.

Unscheduled DNA synthesis (UDS) in the germ cells of male mice after in vivo treatment with X-rays or methyl methanesulfonate (MMS) was assayed by use of a quantitative autoradiographic procedure. MMS induced UDS in meiotic through type III elongating spermatid stages, whereas X-rays induced UDS in meiotic through round spermatid stages. No UDS was detected in the most mature spermatid stages present in the testis with either MMS or X-rays. Taking into account differences in DNA content of the various germ-cell stages studied, we concluded that X-rays induced a maximum UDS response in spermatocytes at diakinesis--metaphase I. The level of UDS induced by MMS was about the same in all the stages capable of repair. Chromosome damage and UDS were measured simultaneously in the same spermatocytes at diakinesis 90 min after X-irradiation or MMS treatment. The level of UDS in most of the X-irradiated cells paralleled the extent of chromosome damage induced. A statistical analysis of these results revealed a positive correlation. As expected, MMS induced no chromosome aberrations above control levels. Therefore no correlation was determined between UDS and chromosome damage in this case. The distribution of UDS over the chromosomes treated at diakinesis with MMS or X-rays was studied. It was found that UDS occurred in clusters in the irradiated cells, whereas it was uniformly distributed in the MMS-treated cells.