The extrachromosomal elements of the Naegleria genus: How little we know.

There are currently 47 characterized species in the Naegleria genus of free-living amoebae. Each amoeba has thousands of extrachromosomal elements that are closed circular structures comprised of a single ribosomal DNA (rDNA) copy and a large non-rDNA sequence. Despite the presence of putative open reading frames and introns, ribosomal RNA is the only established transcript. A single origin of DNA replication (ori) has been mapped within the non-rDNA sequence for one species (N. gruberi), a finding that strongly indicates that these episomes replicate independently of the cell's chromosomal DNA component. This article reviews that which has been published about these interesting DNA elements and by analyzing available sequence data, discusses the possibility that different phylogenetically related clusters of Naegleria species individually conserve ori structures and suggests where the rRNA promoter and termination sites may be located.

The utility of optical coherence tomography for diagnosis of basal cell carcinoma: a quantitative review.

BACKGROUND: Optical coherence tomography (OCT) is a noninvasive near-infrared light imaging technology that can be utilized to diagnose basal cell carcinomas (BCCs) based on specific morphological features. OBJECTIVES: To conduct a quantitative review using tumour-level data from published studies to assess: (i) the in vivo diagnostic accuracy of different OCT systems; (ii) correlation between OCT features and histopathological diagnosis; and (iii) factors that impact the accuracy of tumour depth estimation. METHODS: Primary tumour-level data were extracted from published studies on the use of time-domain (TD-OCT), frequency-domain (FD-OCT) and high-definition (HD-OCT) systems for diagnosis of BCCs. Quality assessment was performed using the Newcastle-Ottawa Scale and the Cochrane Risk of Bias Tool. Sensitivity and specificity for diagnosis of BCC, prevalence of morphological features and correlation of tumour depth between OCT and histopathology were analysed. RESULTS: In total, 901 BCCs from 31 studies were included. The sensitivity and specificity were 89·3% and 60·3% overall, and were highest for FD-OCT (93·7% and 61·4%, respectively). The most prevalent morphological features were lobular pattern (80·2%, 315 of 393 tumours) and hyper-reflective peritumoral stroma (51·7%, 203 of 393). Concordance between OCT and histopathological tumour depth categories was moderate (Pearson coefficient 0·48); it was highest for tumours < 1 mm and those on the extremities. The overall bias was 0·075 mm with an agreement range from -0·88 to 1·03 mm. HD-OCT and FD-OCT were superior to TD-OCT at identifying morphological features, but not at tumour depth estimation. CONCLUSIONS: OCT is a viable tool for in vivo diagnosis of BCCs. FD-OCT and HD-OCT outperformed TD-OCT in diagnostic accuracy and detection of morphological features, but not tumour depth estimation.

Immunization in Vietnam.

Since the Expanded Program on Immunization was proposed by the World Health Organization in 1981, it has been promptly adopted by Vietnam as one of the country's national priority programs. In 1986, Vietnam achieved some remarkable goals, including polio-free status and the elimination of neonatal tetanus. At the same time, however, barriers and difficulties have also emerged. This article aims to provide an overview of both achievements and barriers to the implementation of the program and proposes some solutions.

Soft tissue regeneration in animal models using grafts from adipose mesenchymal stem cells and peripheral blood fibrin gel.

OBJECTIVE: Our study aimed to evaluate the effect of soft tissue regeneration in nude mice using grafts made from the combination of adipocytes from fat tissue mesenchymal stem cells and fibrin gel from peripheral blood. MATERIALS AND METHODS: Mesenchymal stem cells were isolated from adipose tissue and identified according to ISCT criteria. The scaffold used was fibrin obtained from peripheral blood. The grafts in this study were generated by transferring mesenchymal stem cells onto a fibrin scaffold. Two types of grafts, the research sample (fibrin scaffold containing adipocytes differentiated from mesenchymal stem cells) and the control sample (fibrin scaffold only), were grafted under the dorsal skin of the same mouse. After each research period, samples were collected and evaluated by histological methods to observe the existence and growth of cells inside the grafts. RESULTS: The results showed that the study group's graft integrated better within the tissue when compared with the control group. In addition, the grafts in the study group showed the presence of cells with characteristic morphology of adipocytes one week after transplantation. In contrast, control samples showed dimorphous shapes and features mainly composed of non-homogenous fragments. CONCLUSIONS: These initial conclusions might be considered a first step in generating safe bio-compatible engineered grafts specifically usable in post-traumatic tissue regeneration procedures.

A FOXO-dependent replication checkpoint restricts proliferation of damaged cells.

DNA replication is challenged by numerous exogenous and endogenous factors that can interfere with the progression of replication forks. Substantial accumulation of single-stranded DNA during DNA replication activates the DNA replication stress checkpoint response that slows progression from S/G2 to M phase to protect genomic integrity. Whether and how mild replication stress restricts proliferation remains controversial. Here, we identify a cell cycle exit mechanism that prevents S/G2 phase arrested cells from undergoing mitosis after exposure to mild replication stress through premature activation of the anaphase promoting complex/cyclosome (APC/CCDH1). We find that replication stress causes a gradual decrease of the levels of the APC/CCDH1 inhibitor EMI1/FBXO5 through Forkhead box O (FOXO)-mediated inhibition of its transcription factor E2F1. By doing so, FOXOs limit the time during which the replication stress checkpoint is reversible and thereby play an important role in maintaining genomic stability.

Blocks of limited haplotype diversity revealed by high-resolution scanning of human chromosome 21.

Global patterns of human DNA sequence variation (haplotypes) defined by common single nucleotide polymorphisms (SNPs) have important implications for identifying disease associations and human traits. We have used high-density oligonucleotide arrays, in combination with somatic cell genetics, to identify a large fraction of all common human chromosome 21 SNPs and to directly observe the haplotype structure defined by these SNPs. This structure reveals blocks of limited haplotype diversity in which more than 80% of a global human sample can typically be characterized by only three common haplotypes.

Levels of understanding of the rules of correct medical usage among vietnamese pharmacy students: a cross-sectional study.

BACKGROUND: Comprehensive misunderstanding about medicine usage is often associated with high treatment risks which have led to unexpected and adverse effects or even death. Many researches assessing health literacy have been undertaken, but only in adults. This study was undertaken to evaluate the level of understanding in students of medical terms and its correlation with gender, grade and parental occupation. METHODS: A cross-sectional study was conducted from September to October 2017 with 594 students (28.6% of men and 71.4% of women) of Hanoi University of Pharmacy from freshman to fifth-year students chosen randomly. The knowledge of pharmacy students was assessed by a questionnaire including 25 medical terms. Descriptive statistics and Chi-square test were used with p < 0.05 as level of statistical significance. RESULTS: The level of understanding of students was high with most of medical terms reaching over 70% correct answers. A positive significant association between health literacy and education was found with higher knowledge demonstrated in upper years, while there was no difference among students with and without parents belonging to the medical field. Regarding the relation with gender, there was no significant correlation for most medical terms. CONCLUSIONS: Levels of understanding of medical terms in pharmacy students was high, presenting a significant association with education. This study should be extended in order to assess the level of health literacy in various populations, thereby indirect evaluating implementation of medical preventive programs.

Interaction among the distinct effects of adenine and guanine depletion in mouse lymphoma cells.

Toxic guanine depletion was shown previously to result in a dramatic reduction of DNA synthesis, while toxic adenine depletion failed to affect DNA synthesis (M. B. Cohen and W. Sadée, Cancer Res., 43: 1587-1591, 1983). In this study, relative DNA synthesis rates were measured in mouse lymphoma S49 cells over 24 hr after drug exposure and were compared to cell growth curves. DNA synthesis inhibition by mycophenolic acid (guanine starvation) was achieved at lower drug concentrations than was the inhibition of cell growth. This result further supports the hypothesis (reference above) that guanine starvation specifically affects cells in S phase while it allows cells with full DNA complement to divide. In contrast, L-alanosine (adenine starvation) failed to affect DNA synthesis for at least 24 hr at a concentration that inhibits cell growth by 80%. The dramatically different effects of guanine and adenine starvation on DNA synthesis can thus be used to assess the magnitude of each when blocking early de novo purine biosynthesis by 6-methyl-mercaptopurine ribonucleoside (6- MMPR ). The results suggest that, although 6- MMPR effects primarily resemble those of guanine depletion, adenine starvation measurably contributes to the overall toxicity of 6- MMPR . Drug combination experiments with L-alanosine, mycophenolic acid, and 6- MMPR suggest that the basic mechanisms underlying the toxic effects of guanine and adenine starvation act synergistically.

Biochemical differences among four inosinate dehydrogenase inhibitors, mycophenolic acid, ribavirin, tiazofurin, and selenazofurin, studied in mouse lymphoma cell culture.

The mechanism of the cellular toxicity of four inosinate dehydrogenase (IMP-DH) inhibitors with different antitumor and antiviral pharmacological profiles was investigated in mouse lymphoma (S-49) cell culture. Drug effects on cell growth, nucleotide pools, and DNA and RNA synthesis were measured in the presence and absence of guanine salvage supplies. Both guanine and guanosine were capable of bypassing the IMP-DH block, while they also demonstrated some growth-inhibitory effects when added alone in high concentrations. All four drugs reduced cellular guanosine triphosphate levels and caused secondary changes of the uridine, cytidine, and adenosine triphosphate pools that were similar among the four drugs. However, several drug effects in addition to IMP-DH inhibition were observed except with mycophenolic acid which may represent a pure IMP-DH inhibitor. Both tiazofurin and selenazofurin interfered with the uptake and/or metabolism of uridine and thymidine tracers; however, this effect appeared not to contribute to their cellular toxicity in vitro. Moreover, selenazofurin and tiazofurin impaired the utilization of exogenous guanine salvage supplies for DNA and RNA synthesis, and guanine was particularly ineffective in reversing the toxic effects of tiazofurin on cell growth. This finding is important in view of the available guanine salvage supplies in vivo. Since tiazofurin, selenazofurin, and their known metabolites failed to inhibit hypoxanthine-guanine-phosphoribosyl transferase, guanosine monophosphate kinase, and guanosine diphosphate kinase in cell extracts or permeabilized cells, these drugs may interfere with salvage transport across cellular membranes. The toxic effects of mycophenolic acid and ribavirin were similarly reversed by salvage supplies of up to 200 microM guanine, which suggests that ribavirin primarily acts as an IMP-DH inhibitor under these conditions. This result could explain the rather low antitumor efficacy of both mycophenolic acid and ribavirin in vivo. However, increasing the guanine salvage supply in the medium above 200 microM further reversed the toxic effects of mycophenolic acid to maximum rescue, while it increased the toxicity of ribavirin (300 microM). This finding suggests the presence of a toxic mechanism of ribavirin at higher concentrations that is dependent upon the presence of guanine supplies sufficient to fully overcome the IMP-DH inhibition. This study documents that each antimetabolite displays a unique spectrum of activities with multiple toxic targets.

The fate of axonal debris in Wallerian degeneration of rat optic and sciatic nerves. Electron microscopy and immunofluorescence studies with neurofilament antisera.

Immunofluorescence demonstrated that axonal debris reacting with neurofilament antisera persist up to 4 months in rat optic nerves undergoing Wallerian degeneration. Antisera used in this study allowed the isolation of the 72,000- and 150,000-dalton neurofilament polypeptides from rat spinal cord by immunoaffinity chromatography. After 2 weeks of degeneration, proteins co-migrating with these neurofilament polypeptides were no longer identifiable in rat optic nerves, which suggests that immunofluorescent structures persisting in the nerves after this period contained neurofilament degradation products of different molecular weight. Additional evidence as to the persistence of axonal debris in degenerated optic nerves was obtained by electron microscopy. Two distinct types of axonal degeneration were observed in rat optic nerves by this method, floccular swelling and increased electron density of the axoplasm. In both types of degeneration, axoplasmic filaments and tubules were not identifiable. Although floccular material disappeared after 2 weeks of degeneration, so that only empty myelin sheaths remained, electron-dense axons persisted longer and were probably phagocytosed together with their myelin sheaths. In sciatic nerves, cross-reaction with neurofilament antisera had almost completely disappeared 10 days after transection. The same was true for nerves which had been tightly ligated to prevent axonal growth and to squeezed nerves which showed vigorous regeneration. A few scattered, brightly immunofluorescent fragments which persisted in nerves up to 2 weeks after transection were exception to these findings.

Glial fibrillary acidic (GFA) protein in Schwann cells: fact or artifact?

Antisera to the glial fibrillary acidic (GFA) protein stained a subpopulation of Schwann cells in cryostat sections of rat sciatic nerve by indirect immunofluorescence and by the peroxidase-antiperoxidase (PAP) procedure. The staining pattern was entirely different from that obtained with vimentin antisera, which uniformly decorated endoneurial tubes. Electron microscopic examination of sciatic nerve provided a possible explanation for the relatively small number of Schwann cells decorated by GFA antisera: 10 nm filaments were mainly confined to Schwann cell processes surrounding nonmyelinated axons. A marked increase in GFA-positive Schwann cells and in Schwann cells containing filaments by electron microscopy was observed in sciatic nerves undergoing Wallerian degeneration. Conversely, immunochemical procedures failed to demonstrate the presence of antigen reacting with GFA antisera in extracts of sciatic nerve, both normal and degenerated. These include absorption experiments, double immunodiffusion, immunoaffinity chromatography, and immunoradiometric assay. Two explanations may be considered for these findings: i) Schwann cell intermediate filaments and GFA protein share common antigenic determinants, the immunohistological methods being more sensitive to detect cross-reactivity as compared to immunochemical procedures on tissue extracts; and ii) the binding of anti-GFA to Schwann cell 10 nm filaments is not due to immunological cross-reactivity.

Comparative effects of cotrimoxazole (trimethoprim-sulphamethoxazole) and spiramycin in pregnant mice infected with Toxoplasma gondii (Beverley strain).

The effects of cotrimoxazole (CTX) and spiramycin (Spir) in mice infected in midpregnancy with the Beverley (Bev) strain of Toxoplasma gondii were compared. Therapeutic effectiveness was determined according to the following parameters: rate of successful delivery, litter size, offspring weight and survival. When compared with the uninfected untreated control group, CTX showed a more beneficial therapeutic effect than Spir, with a statistically significant increase in the rate of both successful delivery and offspring survival. Results based on antitoxoplasma antibody determinations in the offspring indicated a better in utero control of congenital infection by CTX than by Spir.

Comparative effects of cotrimoxazole (trimethoprim-sulphamethoxazole), pyrimethamine-sulphadiazine and spiramycin during avirulent infection with Toxoplasma gondii (Beverley strain) in mice.

The antitoxoplasm effects of cotrimoxazole (Ctx), spiramycin (Spir) and pyrimethamine-sulphadiazine (Pmm-Sdz) were compared during both proliferative and chronic phases of infection of mice with the Beverley (Bev) strain of Toxoplasma gondii of low virulence. The therapeutic efficacy of the drugs was determined according to the following criteria: (i) specific antibody response; (ii) acquired resistance to lethal challenge with the virulent RH strain of Toxoplasma; and (iii) persistence of parasites in tissues (brain, liver, spleen) of treated mice. The results indicated that Ctx, like Pmm-Sdz, had a greater effect than Spir upon toxoplasma organisms during the proliferative phase of infection. In contrast, none of the three drugs tested was active against tissue cysts in chronically infected mice.

Factors affecting in vitro growth of harvested enterocytes.

Selective enterocyte transplantation may be an alternative to whole organ transplantation for increasing absorptive capacity. Our aim was to determine the effect of initial cell number and viability, proportion of intact crypts, and basement membrane components (BMC) on the in vitro growth of rabbit enterocytes. Enterocytes were harvested using warm trypsinization from ileal segments in 40 rabbits. Initial cell viability was 92 +/- 4% (mean +/- SD), cell yield was 7.7 +/- 3.6 x 10(6) cells/cm, and there were 0.51 +/- 0.33 crypts/100 cells. Initial cell viability correlated with cell yield (r = -0.508, p < 0.001) and % crypts (r = 0.313, p < 0.05). Cell yield also correlated with % crypts (r = -0.645, p < 0.001). Enterocytes (5 x 10(6)) were incubated in growth media in plain or BMC coated growth culture vessels for 14 days. There was a correlation between both number of cells seeded (r = 0.824, p < 0.001) and cell viability (r = -0.696, p < 0.01) and % growth colonies containing epithelial cells at 14 days. Both total growth colonies (r = -0.565, p < 0.05) and colonies with epithelial cells (r = -0.589, p < 0.05) had a negative correlation with % crypts. Incubating cells in BMC coated vessels (n = 6) resulted in significantly more dispase liberated cells after 14 days than in plain vessels (n = 6) (6.6 +/- 1.1 vs. 3.8 +/- 1.0 x 10(6), p < 0.05) but viability was similar (97 +/- 2% vs. 96 +/- 2%).(ABSTRACT TRUNCATED AT 250 WORDS)

Guanine ribonucleotide depletion in mammalian cells. A target of purine antimetabolites.

In a previous report we demonstrated in mouse lymphoma (S-49) cells that DNA synthesis inhibition resulting from guanine starvation is associated with GTP rather than dGTP depletion. Since several effective anticancer drugs act via guanine depletion, it is important to test whether critical GTP depletion is unique to S-49 cells or also occurs in other cell lines. Mycophenolic acid-induced guanine starvation caused a drastic DNA synthesis inhibition in the human lymphoblastic T leukemia (CEM) and the mouse B leukemia (L1210) cell lines, which was again associated with GTP depletion rather than dGTP depletion. These results suggest that GTP depletion represents a common target of purine antimetabolites in mammalian cells.

An association between Vogt-Koyanagi-Harada disease and Guillain-Barré syndrome.

PURPOSE: To describe an association between Vogt-Koyanagi-Harada disease and Guillain-Barré syndrome. METHODS: Case series, describing three patients. RESULTS: In two patients, the disorders had their onsets within 2 weeks of each other; in the third patient, Vogt-Koyanagi-Harada disease occurred after 3 months, as Guillain-Barré syndrome resolved. All three patients had bilateral panuveitis typical of Vogt-Koyanagi-Harada disease. Each also developed well-accepted manifestations of Guillain-Barré syndrome, including paresis of the lower extremities (all patients), paresis of the upper extremities (two patients), paresis of cranial nerves (two patients), areflexia (all patients), and abnormal electromyography findings (two patients). CONCLUSIONS: Vogt-Koyanagi-Harada disease may follow or occur simultaneously with Guillain-Barré syndrome. The fact that these two autoimmune disorders occur together in some patients suggest that they may share common disease mechanisms.

Cervical thorium dioxide granuloma ('thorotrastoma').

An elderly woman had an expanding cervical mass that entrapped and compressed the adjacent cranial nerves, blood vessels, and muscles. The mass was dense on radiographs, extended from the skull base to low neck in the prevertebral and parapharyngeal tissues, and showed mixed intensity on MR. A previous direct carotid arteriogram with thorium dioxide as the contrast agent suggested the histologically proved diagnosis of a cervical thorium dioxide granuloma ("thorotrastoma").

Rationale for the combination of cryoablation with surgical resection of hepatic tumors.

Only 5% to 10% of metastatic and primary liver tumors are amenable to surgical resection. Hepatic cryoablation has increased the number of patients who are suitable for curative treatment. The aim of this study was to evaluate survival and intrahepatic recurrence in patients treated with cryoablation and resection. From June 1994 to July 1999, thirty-eight surgically unresectable patients underwent a total of 42 cryoablative procedures for 65 malignant hepatic lesions. Twenty patients underwent cryoablation alone, and 18 patients were treated with a combination of resection and cryoablation, with a minimum of 18 months' follow-up. The 38 patients had the following malignancies: primary hepatocellular carcinoma (n = 8) and metastases from colorectal cancer (n = 21), neuroendocrine tumors (n = 3), ovarian cancer (n = 3), leiomyosarcoma (n = 1), testicular cancer (n = 1), and endometrial cancer (n = 1). Patients were evaluated preoperatively with spiral CT scans and intraoperatively with ultrasound examinations for lesion location and cryoprobe guidance. Local recurrence was detected by CT. Major complications included bleeding in three patients and acute renal failure, transient liver insufficiency, and postoperative pneumonia in one patient each. Two patients (5%) died during the early postoperative interval; mean hospital stay was 7.1 days. Median follow-up was 28 months (range 18 to 51 months). Overall survival according to Kaplan-Meier analysis was 82%, 65%, and 54% at 12, 24, and 48 months, respectively. Forty-eight-month survival was not significantly different between those patients undergoing cryoablation alone (64%) and those treated with a combination of resection and cryoablation (42%). Disease-free survival at 45 months was 36% for patients undergoing cryoablation plus resection compared to 25% for those undergoing cryoablation alone. Local recurrences were detected at five cryosurgical sites, for a rate of 12% overall (5 of 42), 11% (2 of 18) for patients in the cryoablation plus resection group, and 12% (3 of 24) for those in the cryoablation alone group. For patients with colorectal metastases, survival was 70% at 30 months compared to 33% for hepatocellular cancer and 66% for other types of tumors. Patients with tumors larger than 5 cm or numbering more than three did not have significantly decreased survival. Cryoablation of hepatic tumors is a safe and effective treatment for some patients not amenable to resection. The combination of cryoablation and resection results in survival comparable to that achieved with cryoablation alone.

Human T-lymphotropic Virus Type II Infection inVietnamese Thalassemic Patients.

Anti-human T-lymphotropic virus type I/II (HTLV-I/II) antibodies were screened by particle agglutination test in a total of 66 patients with thalassemia major who received multiple transfusion from paid donors at the Blood Transfusion Hematology Center of Ho Chi Minh City in South Vietnam. HTLV-II infection was confirmed in 6 patients (9.1%) by Western blot analysis and/or polymerase chain reaction. Phylogenetic analysis revealed that long terminal repeat sequences of HTLV-II proviruses from 5 thalassemic patients in Vietnam belonged to the same phylogenetic subgroup of HTLV-IIb as those from intravenous drug abusers in North America and Europe. These data shed light on the route of introducing HTLV-II into Vietnam.

Should intestinal continuity be restored after massive intestinal resection?

We reviewed our experience with 32 patients who underwent massive intestinal resection to determine when intestinal continuity should be maintained. An enterostomy was created in 21 patients (66 percent) at the initial resection because of questionable bowel viability and an unstable condition or the need for colonic anastomosis. Intestinal continuity was restored in only 20 percent of these patients. In 11 patients (34 percent), intestinal continuity was maintained at the time of resection. Only four of these patients (36 percent) had a satisfactory long-term outcome. Overall, intestinal continuity was maintained in 10 of the 22 patients (45 percent) followed over the long-term. Three quarters of patients with intestinal remnants shorter than 3 feet had an enterostomy. We believe intestinal continuity should be restored at the time of massive resection only in carefully selected patients when bowel viability is ensured, remnant length is greater than 3 feet, and a colonic anastomosis is not required. Maintaining intestinal continuity eliminates the inconvenience of the stoma but may cause dietary restriction and perianal discomfort.