CARDIOVASCULAR RISK FACTORS IN ADOLESCENTS: ROLE OF INSULIN RESISTANCE AND OBESITY.

INTRODUCTION: Childhood obesity is a public health problem characterized by early insulin resistance (IR), inflammation, and oxidative stress. The presence of an uninterrupted low-grade inflammatory state impairs metabolic and cardiovascular health. The population is particularly susceptible to develop metabolic disorders related to increased body fat. METHODS: Eighty-three adolescents were recruited and grouped according to HOMA-IR and BMI in either with or without IR and obese or normal-weight respectively. Anthropometric, biochemical, immunological and hormonal variables were determined. Transverse Analytical Study. RESULTS: Obesity, dyslipidemia, IL-6, and C-reactive protein were significantly higher in the IR group than in the non-IR group. Obese adolescents showed increased insulin levels, HOMA-IR, inflammatory markers, and triglycerides; while having lower HDL-C, and adiponectin when compared to normal-weight adolescents. As expected, obesity-related anthropometric markers positively correlated with IR and inflammatory markers while negatively correlated with adiponectin levels. CONCLUSIONS: Early IR, subclinical inflammation, dyslipidemia, and hypoadiponectinemia characterize obesity in adolescents. These factors may increase the risk of future coronary heart disease (CHD) and diabetes mellitus development (DM) in early adulthood.

Bioenergetic stress potentiates antimicrobial resistance and persistence.

Antimicrobial resistance (AMR) is a global health crisis and there is an urgent need to better understand AMR mechanisms. Antibiotic treatment alters several aspects of bacterial physiology, including increased ATP utilization, carbon metabolism, and reactive oxygen species (ROS) formation. However, how the "bioenergetic stress" induced by increased ATP utilization affects treatment outcomes is unknown. Here we utilized a synthetic biology approach to study the direct effects of bioenergetic stress on antibiotic efficacy. We engineered a genetic system that constitutively hydrolyzes ATP or NADH in Escherichia coli. We found that bioenergetic stress potentiates AMR evolution via enhanced ROS production, mutagenic break repair, and transcription-coupled repair. We also find that bioenergetic stress potentiates antimicrobial persistence via potentiated stringent response activation. We propose a unifying model that antibiotic-induced antimicrobial resistance and persistence is caused by antibiotic-induced. This has important implications for preventing or curbing the spread of AMR infections.

Fanconi-like anemia related to a FANCM mutation.

Fanconi anemia is primarily inherited as an autosomal recessive genetic disorder with common delays in diagnosis and challenging treatments. Fanconi anemia patients have a high risk of developing solid tumors, particularly in the head and neck or anogenital regions. The diagnosis of Fanconi anemia is primarily based on the chromosomal breakage but FA gene sequencing is recommended in all patients with a positive chromosome fragility test. Here, we present a 32-year-old man with advanced tonsil squamous cell carcinoma and fatal toxicity after the first cycle of chemotherapy. No anemia was present. A recent variant mutation if the FANCM gene was detected (c1511_1515delGAGTA (pArg504AsnfsTer29)). Homozygous or double heterozygous pathogenic variants have been reported in FANCM and linked to azoospermia and primary ovarian failure without anemia. Alterations in this gene have also been associated with a genetic predisposition for solid tumors (breast and ovarian cancer) and hematological malignancies (B-cell acute lymphoblastic leukemia). Due to the hypersensitivity of these patients to DNA-damaging agents such as chemotherapy and radiotherapy, surgery is the best treatment option for malignant solid tumors. Dose reductions or alternative regimens of chemotherapy and/or radiotherapy are recommended in FA patients who develop a malignant tumor.

Studying semi-dynamic digestion kinetics of food: Establishing a computer-controlled multireactor approach.

In this work, a multireactor system to study digestion (MuReDi) kinetics is introduced. For this, a custom-made automated system with four independent syringe pumps (BioXplorer 100, H.E.L Group) was acquired. This system consists of multiple, small-scale reactors allowing to study digestion as a function of time and thus to determine digestion kinetics. The different digestion conditions used in the oral, gastric, and small intestinal phase were based on the digestion protocols published by the INFOGEST consortium. We showed that the minimum working volume of a reactor is 30 mL. Besides, repeatability of the digestion kinetics was shown for two food systems: a liquid Ensure® Plus Vanilla drink, and a solid, cooked lentil sample. When comparing static digestion kinetics with semi-dynamic ones, a significantly different digestion pattern was observed. In the static case, a relatively fast hydrolysis rate was observed until a clear plateau was reached. Oppositely, for the semi-dynamic case, a delayed start of the hydrolysis process was noticed. In the gastric phase, this was explained by the decreasing pH and the large pH dependency of pepsin activity. In the small intestine, the lag phase was relatively shorter, yet clearly present. Here we related it to the gradual enzyme (and bile salt) secretion that had to diffuse towards the substrate before hydrolysis could start. Generally, this work showed that the MuReDi system could be used to perform a semi-dynamic digestion approach which largely impacted the overall digestion kinetics. This is important to consider in future in vitro food digestion simulation work to come closer to physiologically relevant digestion kinetics.

Digestion kinetics of lipids and proteins in plant-based shakes: Impact of processing conditions and resulting structural properties.

In this work, plant-based shakes were prepared (5% oil, 6% protein, 1% lecithin, 88% water) (w/w) using two processing techniques (i) only mixing versus (ii) mixing followed by high pressure homogenisation, as well as two processing sequences (i) adding all ingredients together versus (ii) stepwise addition of ingredients. Shakes only mixed consisted of large, irregular particles (1-100 µm). Eventually, this resulted in a relatively low lipid and protein digestion extent after 2 h of gastric pre-digestion (9% and < 1%, respectively). In contrast, shakes that were subjected to high pressure homogenisation displayed small, homogeneous particles (<10 µm). Besides, lipids and proteins were digested to a high extent in the stomach (40% and 10%, respectively). The small intestinal digestion kinetics indicated a significant impact of proteins on lipid digestion kineticsbutno significant effect of lipids on protein digestion kinetics. The results highlighted the relevance of food processing on macronutrient (micro)structure and further gastrointestinal functionality.

Cryptococcuria as manifestation of disseminated cryptococcosis: Staib agar as a selective identification medium.

We conducted a retrospective study of 58 cases of cryptococcosis (1986-2008) with urine test positive for Cryptococcus sp, in Mycology Laboratory, Santa Casa-Hospital Complex, Porto Alegre, RS, Brazil. The diagnosis of cryptococcuria was based on microscopic examination and culture of urinary sediment. Cryptococcus was isolated from other clinical specimens such as blood, cerebrospinal fluid, ascitic and pleural fluids, respiratory secretions, biopsies of skin, nasal and bone marrow. Cryptocccus neoformans was present in 55 cases and Cryptocccus gattii in three cases. Males predominated (79.3%); age ranged from 12 to 86 years. Acquired Immune Deficiency Syndrome (AIDS) were present in 60.3%, 31.1% did not have AIDS and 5.2% were apparently immunocompetent patients. The most frequent signs and symptoms were headache (53.4%) and fever (51.7%). The most widely used medication was the amphotericin B (43 patients). The mortality rate was 45%. We conclude that the mycological examination of the urine can be an alternative simple, non-invasive and useful in diagnosis of disseminated cryptococcosis, especially when used in conjunction with techniques for demonstration of the capsule (nigrosine) and/or production of melanin in special culture media (Staib agar).

Utility of capsule endoscopy in the diagnosis of gastrointestinal graft-versus-host disease.

INTRODUCTION AND AIM: Graft-versus-host disease (GvHD) is a complication of hematopoietic cell transplantation, and the small bowel is one of the main targets in the gastrointestinal tract. Capsule endoscopy is a safe procedure and can be useful in the diagnosis of GvHD. The aim of the present study was to compare the diagnostic yield of capsule endoscopy with the histopathologic findings in GvHD. MATERIALS AND METHODS: A retrospective diagnostic test study included all the patients with suspected GvHD that underwent gastroscopy and colonoscopy, with histopathologic evaluation of the biopsies taken, and capsule endoscopy, within the time frame of July 2015 and July 2019. Capsule endoscopy findings were compared with the histopathologic diagnosis, considered the gold standard. RESULTS: Twenty-one patients with GvHD (7 [33%] women; 37 ± 11.9 years of age) were included, 20 (95%) of whom had acute GvHD. The median gastric transit time of the capsule was 55 minutes (20-113) and the median small bowel transit time was 261 minutes (238-434). The entire small bowel was visualized through capsule endoscopy in 17 cases (80.95%). The histopathologic findings and capsule endoscopy findings resulted in the diagnosis of GvHD in 17 and 16 cases, respectively. There was agreement between the histopathologic and capsule endoscopy findings in 18 cases (15 positive and 3 negative). Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic yield of capsule endoscopy were 88%, 75%, 94%, 60%, and 85%, respectively. CONCLUSIONS: Capsule endoscopy is a safe tool for the diagnosis of GvHD, with high sensitivity and positive predictive value, as well as moderate agreement with histopathologic findings.

Differential analysis of the bacterial community in colostrum samples from women with gestational diabetes mellitus and obesity.

Gestational Diabetes Mellitus (GDM) and obesity affect the functioning of multiple maternal systems and influence colonization of the newborn gastrointestinal through the breastmilk microbiota (BMM). It is currently unclear how GDM and obesity affect the human BMM composition. Here, we applied 16S-rRNA high-throughput sequencing to human colostrum milk to characterize BMM taxonomic changes in a cohort of 43 individuals classified in six subgroups according to mothers patho-physiological conditions (healthy control (n = 18), GDM (n = 13), or obesity (n = 12)) and newborn gender. Using various diversity indicators, including Shannon/Faith phylogenetic index and UniFrac/robust Aitchison distances, we evidenced that BMM composition was influenced by the infant gender in the obesity subgroup. In addition, the GDM group presented higher microbial diversity compared to the control group. Staphylococcus, Corynebacterium 1, Anaerococcus and Prevotella were overrepresented in colostrum from women with either obesity or GDM, compared to control samples. Finally, Rhodobacteraceae was distinct for GDM and 5 families (Bdellovibrionaceae, Halomonadaceae, Shewanellaceae, Saccharimonadales and Vibrionaceae) were distinct for obesity subgroups with an absolute effect size greater than 1 and a q-value ≤ 0.05. This study represents the first effort to describe the impact of maternal GDM and obesity on BMM.

Effects of acute fasting and age on leptin and peroxisome proliferator-activated receptor gamma production relative to fat depot in immature and mature pigs.

Leptin and peroxisome proliferator-activated receptor gamma (PPARγ) are adipogenic proteins that are actively involved in metabolic homeostasis of fat. Recently, it was reported that fat tissue in humans and rodents differs in metabolic activity relative to anatomical location of the fat tissue (i.e. depots) and animal age. Hence, we hypothesized that leptin and PPARγ production in various fat depots in female pigs differs in response to acute fasting, and that these responses vary with physiological maturity of the animal. Sixteen intact crossbred immature female pigs [prepubertal (PP); 132.2 ± 4.1 days] and 16 sexually mature female pigs (M; 224 ± 7.4 days) housed in an open-air, concrete slab, sheltered barn were randomly assigned to either Control or Fasted treatments. Control pigs (PP, n = 8; M, n = 8) had ad libitum access to feed, while Fasted pigs (PP, n = 8; M, n = 8) were denied access to feed from the onset of the study (0 h) to euthanasia at 72 h. Immediately post-mortem, fat samples were collected from the subcutaneous, pelvic, kidney, and heart (M pigs only) fat depots and analysed for leptin and PPARγ mRNA and protein content. Acute fasting decreased mean leptin mRNA tissue content in a depot specific manner in M pigs (p < 0.01), while mean leptin protein concentrations in fat tissues did not differ with fat depot or age of the pig. Furthermore, acute fasting did not affect mean PPARγ mRNA tissue content in a fat depot or age dependent manner. Mean concentrations of PPARγ protein in fat depots tended to be greater in M vs. PP pigs (p = 0.07). We suggest that these data provide evidence that acute fasting has a greater effect on leptin than PPARγ production in a fat depot dependent manner in M pigs, which may be indicative of changing physiological demands as an animal matures.

Enzymatic and chemical conversions taking place during in vitro gastric lipid digestion: The effect of emulsion droplet size behavior.

This investigation reports the effect of droplet size behavior on the overall lipolysis profile and molecular lipolysis mechanisms under in vitro gastric conditions. O/W emulsions (5% triolein, 1% sodium taurodeoxycholate) with different initial droplet sizes (fine: 0.58 µm; medium: 1.82 µm; and large: 4.00 µm) were subjected to static in vitro digestion. For the first time, multiple lipolysis products including diolein and monoolein regioisomers were quantified within a single HPLC run. An inverse relation was found between the droplet size and the initial rate and final extent of lipolysis based on the digested triolein. Furthermore, a mechanistic gastric lipolysis model was established based on a reaction scheme including enzymatic and chemical isomerization conversions. The estimated rate of the sn-1/3 hydrolysis was around two- to thirty-fold faster compared to the rates of sn-2 cleavage and isomerization, respectively. These findings resulted in a profound insight in in vitro gastric molecular lipolysis mechanisms.

Modulation of CYP2E1 metabolic activity in a cohort of confirmed caffeine ingesting pregnant women with preterm offspring.

BACKGROUND: To ascertain interactions of caffeine ingestion, food, medications, and environmental exposures during preterm human gestation, under informed consent, we studied a cohort of Mexican women with further preterm offspring born at ≤ 34 completed weeks. At birth, blood samples were taken from mothers and umbilical cords to determine caffeine and metabolites concentrations and CYP1A2 (rs762551) and CYP2E1 (rs2031920, rs3813867) polymorphisms involved in caffeine metabolism. RESULTS: In 90 pregnant women who gave birth to 98 preterm neonates, self-informed caffeine ingestion rate was 97%, laboratory confirmed rate was 93 %. Theobromine was the predominant metabolite found. Consumption of acetaminophen correlated significantly with changes in caffeine metabolism (acetaminophen R2 = 0.637, p = 0.01) due to activation of CYP2E1 alternate pathways. The main caffeine source was cola soft drinks. CONCLUSION: Environmental exposures, especially acetaminophen ingestion during human preterm pregnancy, can modulate CYP2E1 metabolic activity.

Ossification of the phalanges of the foot and its relationship to peak height velocity and the calcaneal system.

PURPOSE: There are multiple skeletal maturity grading systems, but none of them utilizes the phalanges of the foot. To minimize radiation, it would be ideal if one could assess the skeletal maturity of a foot based on bones seen on routine foot radiographs, if guided growth is being considered as a treatment option. We developed a system that correlates changes of the appearance of the foot phalanges to peak height velocity (PHV) and the recently described calcaneal apophyseal ossification grading system. METHODS: We selected 94 children from the Bolton-Brush study, each with consecutive radiographs from age ten to 15 years old. Using the anteroposterior view, we analyzed the ossification patterns of the phalanges and developed a six-stage system. We then determined the PHV for each subject and defined its relationship with our system. Our system was then compared with the previously established calcaneal system. RESULTS: We calculated an Intraclass correlation coefficient (ICC) range of 0.957 to 0.985 with a mean of 0.975 and interclass reliability coefficient of 0.993 indicating that this method is reliable and consistent. Our system showed no significant difference between gender with respect to PHV, which makes it a reliable surrogate for determining bone age in paediatric and adolescent patients. CONCLUSIONS: Our system has a strong association with the calcaneal system. It is a simple six-stage system that is reliable and correlated more strongly with PHV than chronological age. The system requires knowledge of the ossification markers used for each stage but is easily used in a clinical setting.

Phenotype and natural history of elderly onset inflammatory bowel disease: a multicentre, case-control study.

BACKGROUND: Onset during old age has been reported in upto 10% of total cases of inflammatory bowel disease (IBD). AIM: To evaluate phenotypic characteristics and the use of therapeutic resources in patients with elderly onset IBD. METHODS: Case-control study including all those patients diagnosed with IBD over the age of 60 years since 2000 who were followed-up for >12 months, identified from the IBD databases. Elderly onset cases were compared with IBD patients aged 18 to 40 years at diagnosis, matched by year of diagnosis, gender and type of IBD (adult-onset). RESULTS: One thousand three hundred and seventy-four elderly onset and 1374 adult-onset cases were included (62% ulcerative colitis (UC), 38% Crohn's disease (CD)). Among UC patients, elderly onset cases had a lower proportion of extensive disease (33% vs 39%; P < 0.0001). In CD, elderly onset cases showed an increased rate of stenosing pattern (24% vs 13%; P < 0.0001) and exclusive colonic location (28% vs 16%; P < 0.0001), whereas penetrating pattern (12% vs 19%; P < 0.0001) was significantly less frequent. Regarding the use of therapeutic resources, there was a significantly lower use of corticosteroids (P < 0.0001), immunosuppressants (P < 0.0001) and anti-TNFs agents (P < 0.0001) in elderly onset cases. Regarding surgery, we found a significantly higher surgery rate among elderly onset UC cases (8.3% vs 5.1%; P < 0.009). Finally, elderly onset cases were characterised by a higher rate of hospitalisations (66% vs 49%; P < 0.0001) and neoplasms (14% vs 0.5%; P < 0.0001). CONCLUSIONS: Elderly onset IBD shows specific characteristics and they are managed differently, with a lower use of immunosuppressants and a higher rate of surgery in UC.

Comparison of cytotoxicity and genotoxicity effects of silver nanoparticles on human cervix and breast cancer cell lines.

The wide application of silver nanoparticles (AgNPs) has pointed out the need to evaluate their potential risk and toxic effects on human health. Herein, the cytotoxic effects of Argovit™ AgNPs were evaluated on eight cancer cell lines. Further cytotoxic studies were performed in gynecological cancer cell lines from cervical (HeLa) and breast (MDA-MB-231 and MCF7) cancer. In both cases, the half maximal inhibitory concentration (IC50) of AgNPs produced the formation of reactive oxygen species (ROS) after 24 h of incubation, but it was not statistically significant compared with untreated cells. However, HeLa, MDA-MB-231, and MCF7 cells treated with the maximal IC of AgNPs induced the formation of ROS either at 12 or 24 h of incubation. Genotoxicity achieved by comet assay in HeLa, MDA-MB-231, and MCF7 cells revealed that exposure to IC50 of AgNPs does not induced noticeable DNA damage in the cells. However, the IC of AgNPs provoked severe DNA damage after 12 and 24 h of exposure. We conclude that, Argovit (polyvinylpyrrolidone-coated AgNPs) induce a cytotoxic effect in a time and dose-dependent manner in all the eight cancer cell lines tested. Nevertheless, the genotoxic effect is mainly restricted by the concentration effect. The results contribute to explore new therapeutic applications of AgNPs for malignances in murine models and to study in deep the cytotoxic and genotoxic effects of AgNPs in healthy cells at the surrounding tissue of the neoplasia.

PAX7 is a required target for microRNA-206-induced differentiation of fusion-negative rhabdomyosarcoma.

Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood. RMS can be parsed based on clinical outcome into two subtypes, fusion-positive RMS (FP-RMS) or fusion-negative RMS (FN-RMS) based on the presence or absence of either PAX3-FOXO1 or PAX7-FOXO1 gene fusions. In both RMS subtypes, tumor cells show histology and a gene expression pattern resembling that of developmentally arrested skeletal muscle. Differentiation therapy is an attractive approach to embryonal tumors of childhood including RMS; however, agents to drive RMS differentiation have not entered the clinic and their mechanisms remain unclear. MicroRNA-206 (miR-206) expression increases through normal muscle development and has decreased levels in RMS compared with normal skeletal muscle. Increasing miR-206 expression drives differentiation of RMS, but the target genes responsible for the relief of the development arrest are largely unknown. Using a combinatorial approach with gene and proteomic profiling coupled with genetic rescue, we identified key miR-206 targets responsible for the FN-RMS differentiation blockade, PAX7, PAX3, NOTCH3, and CCND2. Specifically, we determined that PAX7 downregulation is necessary for miR-206-induced cell cycle exit and myogenic differentiation in FN-RMS but not in FP-RMS. Gene knockdown of targets necessary for miR-206-induced differentiation alone or in combination was not sufficient to phenocopy the differentiation phenotype from miR-206, thus illustrating that miR-206 replacement offers the ability to modulate a complex network of genes responsible for the developmental arrest in FN-RMS. Genetic deletion of miR-206 in a mouse model of FN-RMS accelerated and exacerbated tumor development, indicating that both in vitro and in vivo miR-206 acts as a tumor suppressor in FN-RMS at least partially through downregulation of PAX7. Collectively, our results illustrate that miR-206 relieves the differentiation arrest in FN-RMS and suggests that miR-206 replacement could be a potential therapeutic differentiation strategy.

[Identification of auditory laterality by means of a new dichotic digit test in Spanish, and body laterality and spatial orientation in children with dyslexia and in controls]. / Identificación de la lateralidad auditiva mediante una prueba dicótica nueva con dígitos en español, y de la lateralidad corporal y orientación espacial en niños con dislexia y en controles.

INTRODUCTION: In this study, a new dichotic digit test in Spanish (NDDTS) was applied in order to identify auditory laterality. We also evaluated body laterality and spatial location using the Subirana test. SUBJECTS AND METHODS: Both the dichotic test and the Subirana test for body laterality and spatial location were applied in a group of 40 children with dyslexia and in a control group made up of 40 children who were paired according to age and gender. The results of the three evaluations were analysed using the SPSS 10 software application, with Pearson's chi-squared test. RESULTS: It was seen that 42.5% of the children in the group of dyslexics had mixed auditory laterality, compared to 7.5% in the control group (p < or = 0.05). Body laterality was mixed in 25% of dyslexic children and in 2.5% in the control group (p < or = 0.05) and there was 72.5% spatial disorientation in the group of dyslexics, whereas only 15% (p < or = 0.05) was found in the control group. CONCLUSIONS: The NDDTS proved to be a useful tool for demonstrating that mixed auditory laterality and auditory predominance of the left ear are linked to dyslexia. The results of this test exceed those obtained for body laterality. Spatial orientation is indeed altered in children with dyslexia. The importance of this finding makes it necessary to study the central auditory processes in all cases in order to define better rehabilitation strategies in Spanish-speaking children.

Thyroid stimulating hormone response to thyrotropin in prepubertal depression.

In an effort to evaluate whether differences exist in the hypothalamic-pituitary-thyroid axis of depressed children, a thyrotropin releasing hormone (TRH) stimulation test was administered to 55 prepubertal subjects who were divided into three groups matched for age and sex: a depressed group (endogenous N = 15, nonendogenous N = 15), a psychiatric nondepressed control group (N = 16), and a normal control group (N = 9). Each subject was tested at two dosages of TRH, 2 micrograms/kg and 7 micrograms/kg. Increasing age and female sex were positively correlated with a greater thyroid stimulating hormone (TSH) response. TSH response to TRH was examined with subjects reclassified by severe suicidal ideation, severe aggression, and parental history of alcoholism. Results of this study are contrasted with the adult psychiatric literature.

Cellular immunity in depressed, conduct disorder, and normal adolescents: role of adverse life events.

OBJECTIVE: To determine whether adolescents with major depressive disorder have disturbances in their cellular immunity and to study whether the immunological changes detected are specific to depression or are general responses to stress. METHOD: Twenty subjects with major depressive disorder, 17 nondepressed subjects with conduct disorder, and 17 normal adolescents were recruited. Subjects were assessed with a clinical interview for DSM-III-R and a modified version of the Coddington Life Events Checklist. Blood samples were drawn for total white blood cells, lymphocytes subsets, natural killer cell activity, lymphocyte proliferation response to phytohemagglutinin, and cortisol plasma levels. RESULTS: Overall, there were no significant between-group differences in any of the cellular immune measurements. Natural killer cell activity was significantly negatively correlated with past year and lifetime adverse life events across all effector-target cell ratios. Controlling for diagnoses and socioeconomic status yielded similar results. There were no significant effects of age, sex, race, sleep, nutrition, cigarette use, menstrual cycle, or cortisol on any of the immunological variables. CONCLUSIONS: In this sample of adolescents, we found that independent of the diagnoses and socioeconomic status, increases in adverse life events were associated with low natural killer cell activity.

Embryo cryopreservation in cynomolgus monkeys.

An improved knowledge of cryopreservation of primate embryos will have important research and clinical application. Fifty-six 4- to 8-cell in vitro fertilized embryos were frozen in HEPES-buffered Tyrode's solution containing 1.5 M dimethylsulfoxide (DMSO) and cooled at the rate of 0.3 degrees C/minute to -39 degrees C before being transferred into liquid nitrogen. Embryos were rapidly thawed at room temperature for 2 minutes. DMSO was diluted with medium in three steps at 5-minute intervals. Of the 56 embryos, 39 (70%) were classified as viable on the basis of surviving the freezing process with greater than 50% of their blastomeres intact. Twelve of the 39 embryos were cultured overnight, and 11 cleaved at least once. Twenty-five embryos were transferred to nine synchronized, unstimulated recipient monkeys 24 to 48 hours after ovulation. Three pregnancies (33.3%) resulted from the nine transfers.

Nitazoxanide for the treatment of intestinal protozoan and helminthic infections in Mexico.

A study to evaluate the efficacy and safety of nitazoxanide as a single agent for the treatment of a broad spectrum of mixed parasitic infections, both protozoa and helminths, was conducted at a primary school in San Pedro Tolimán, Querétaro, Mexico. Three faecal samples from 1824 adults and children were screened for the presence of oocysts, cysts, trophozoites, eggs or larvae of intestinal protozoa or helminths. Two hundred and forty-six adults and children infected with at least one protozoan and 2 helminths were given 7.5 mg/kg of nitazoxanide (500 mg to adults and 200 mg to children less than 12 years old) every 12 h for 3 consecutive days. Faecal samples were examined on days 6, 7, 8, 13, 14 and 15 (+/- 1) following initiation of treatment, using formalin-ether concentration and Kato-Katz egg counting. Treatment with nitazoxanide was 71-100% effective in eliminating evidence of infection with Entamoeba histolytica/E. dispar, Giardia duodenalis, Blastocystis hominis, Isospora belli, Enterobius vermicularis, Ascaris lumbricoides, Trichuris trichiura and Hymenolepis nana. Haematology and clinical chemistry values obtained before and after treatment remained unaffected by nitazoxanide. The drug was well tolerated, with only 15 patients (6.1%) reporting mild abdominal pain that lasted less than 24 h.