Sex hormones, SHBG and cognitive performance among older Australian women: an observational study.

OBJECTIVE: This study aims to explore the associations between sex hormones and cognitive performance in older women. METHODS: Associations between sex hormones, sex hormone binding globulin (SHBG) and cognitive performance were examined in women aged at least 70 years, without dementia and not using medications that influence sex hormones. Linear and generalized linear regression models included age, body mass index, education, smoking, alcohol, living circumstances, diabetes, hypertension, depression and impaired renal function. RESULTS: The included 5511 women had a median (interquartile range) age of 73.9 (71.6-77.6) years. No associations were found for estrone, estradiol, testosterone or dehydroepiandrosterone and cognitive performance. SHBG concentrations above quartile 1 (Q1) were significantly inversely associated with processing speed (Q2, ß = -0.94, 95% confidence interval [CI] - 1.64 to -0.24, p = 0.009; Q3, ß = -0.82, 95% CI -1.53 to -0.10, p = 0.025; and Q4, ß = -0.95, 95% CI -1.70 to -0.20, p = 0.013). CONCLUSIONS: Sex hormones were not associated with cognitive performance. The finding that low SHBG is associated with better processing speed warrants further investigation. The null findings for the sex hormones establish a firm baseline to confidently explore the association between sex hormones and longitudinal cognitive performance in this population. TRIAL REGISTRATION: International Standard Randomized Controlled Trial Number Register (ISRCTN83772183) and ClinicalTrials.gov (NCT01038583).

Association between testosterone and cognitive performance in postmenopausal women: a systematic review of observational studies.

This review was conducted to explore the association between endogenous testosterone blood concentrations and cognitive performance among community dwelling postmenopausal women. We searched Ovid MEDLINE, EMBASE, PsycINFO and Web of Science databases for observational studies with at least 100 postmenopausal participants. The results were categorized by study design, reporting of total or free testosterone and risk of bias assessments, narratively. Ten of the 26 articles retrieved for full-text review met the inclusion criteria, six provided cross-sectional data, seven provided longitudinal data and one provided case-control data. Cognitive performance tests differed between studies. Eight studies measured testosterone by immunoassay, one by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and one did not specify their methodology. Eleven different cognitive domains were tested by 37 different instruments. Irrespective of the study design, the findings were inconsistent and inconclusive. Both positive and inverse associations were reported for each of global cognition and immediate and delayed verbal recall. The majority of studies reported no association between total or free testosterone and cognitive performance. Although this review did not demonstrate an association between testosterone and cognitive performance in postmenopausal women, the findings should be considered inconclusive due to the imprecision of testosterone measurement and the methodological heterogeneity of the included studies.

Analytical performance of HPV assays on vaginal self-collected vs practitioner-collected cervical samples: the SCoPE study.

BACKGROUND: In the last decade, human papillomavirus (HPV) testing has been evaluated extensively for cervical screening, with studies finding increased sensitivity compared to cytology. Another advantage of HPV based-screening is the ability to test vaginal samples that can be collected by women themselves. Self-collection has the potential to extend cervical screening coverage by increasing participation rates, particularly among women who are under-screened or have never screened. This could have a significant impact on cervical cancer prevention, as the majority of invasive cervical cancer cases occur among under-screened women. Both the Netherlands and Australia have transitioned their national programs from cytology to HPV as the primary screening test and both countries include a pathway for self-collection. OBJECTIVES: We evaluated the relative sensitivity for HPV detection of self-collection compared with practitioner-collected cervical specimens in the context of the Australian National Cervical Screening Program (NCSP). STUDY DESIGN: 303 women aged ≥18 years attending a single tertiary referral centre took their own sample using a flocked-swab, and then had a practitioner-collected sample taken at colposcopy. All samples were tested at a single laboratory on the six PCR-based HPV assays which can be utilised in the NCSP; Roche cobas 4800 and cobas, Abbott RealTime, BD Onclarity, Cepheid Xpert, and Seegene Anyplex. RESULTS: HPV16/18 results had high observed agreement between self- and practitioner-collected samples on all assays (range: 0.94-0.99), with good agreement for non-HPV16/18 oncogenic HPV types (range: 0.64-0.73). CONCLUSIONS: Self-collection for HPV-based cervical screening shows good concordance and relative sensitivity when compared to practitionercollected samples across assays in the NCSP.

Environmental Situation in Some Selected Villages of Bangladesh.

Health is multifactorial and influenced by nutritional, biological, chemical and psychological factors. It is quite true that environment has direct impact on those diseases that are the outcome of man's maladjustments to his environment. Often man is responsible for pollution of environment through urbanization, industrialization and other human activities. Improvement of environmental condition is one of the best ways to prevent some of the diseases prevailing in our country. The study was designed to explore the environmental situation of rural area of Bangladesh. The descriptive cross sectional study was conducted in purposively selected six villages of Dewanganj Upazila of Jamalpur district, Bangladesh during the period of January 2018 to February 2018. A total of 227 purposively selected women having at least one under five children were interviewed face to face using a semi-structured questionnaire. Mean±SD age of the respondents was 27.31±6.4 years. About three fourth (167, 73.57%) of the respondents were literate while one fourth (60, 26.43%) of them were illiterate. On the other hand about two third (149, 65.64%) of husbands were literate and remaining (78, 34.36%) were illiterate. The most (215, 94.71%) of the respondents were housewives. Majority (143, 62.99%) of them had children of less than 3 and mean±SD age of children was 2.85±1.17 years. Majority (140, 61.67%) of the respondents belonged to nuclear family while 86 (37.89%) belonged to joint family. Mean±SD family member was 6.42±8.12 persons. The mean monthly family income was BDT 8066.52 with a standard deviation of BDT. 6767.12. Out of 277 respondents, almost half 110(48.46%) of them lived in mixed houses followed by 92(40.53%) and 25(11.01%) lived in Kantcha & Pucca houses respectively. The most (180, 79.30%) of the respondents were used separate kitchen in their houses. About two forth (171, 75.33%) of the respondents did not have any kitchen garden. All 227(100.0%) of the respondents were used tube well as source of drinking water. The most (211, 92.95%) of the respondents got supply water within their compounds. All of the respondents got water supply within 15 minute walking distance. Majority 149(65.64%) of the respondents used sanitary toilets & 40(17.62%) used pit latrines and rest 30(13.22%) of them practiced open air disposal. More than half (118, 51.99%) of the respondents' children were not using toilets for excreta disposal. Efforts are needed to increase environmental health related knowledge and awareness to maintain environment sanitation. People should use sanitary latrines and avoid practice of open air disposal of children's feces to prevent excreta borne diseases. Further wider scale details study is needed to formulate policy in regards to improve the environmental sanitation of Bangladesh.

Irritable Bowel Syndrome: Is it Rare in Children?

Irritable bowel syndrome (IBS) is one of the most common and best studied disorders among the group of functional gastrointestinal disorders. It is a functional bowel disorder in which abdominal pain or discomfort is associated with defecation or a change in bowel habit. Visceral hypersensitivity and increased GIT motility are the main patho-physiological mechanism for developing IBS. IBS present with diarrhea and constipation or both. Investigation is least needed for diagnosis of IBS rather done to exclude differential diagnosis. Diagnosis is done on the basis of Rome-III criteria. Proper counseling, dietary management, anti-spasmotic and antidepressant are the mainstay of treatment.

Clinical and Laboratory Predictors of Esophageal Varices in Children with Chronic Liver Disease.

Esophageal varices are a serious consequence of portal hypertension in patients with chronic liver diseases. Several studies have evaluated possible noninvasive markers of esophageal varices to reduce the number of unnecessary endoscopies in patients with chronic liver disease. Aim of this study was to identify clinical and laboratory predictors of esophageal varices in children with chronic liver disease. This cross sectional observational study was done at Pediatric Gastroenterology and Nutrition Department of Bangabandhu Sheikh Mujib Medical University, Dhaka, over a period of 6 months. Fifty (50) consecutive cases of chronic liver disease patient under 15 years of age were evaluated clinically, biochemically and endoscopically. Esophageal varices were classified into 4 grades according to Conn's grading system. Based on endoscopic findings (presence/absence of esophageal varices) all the patients were categorized into two groups; Group I: Chronic liver disease with esophageal varices and Group II: Chronic liver disease without esophageal varices. Most (51.5%) of the patients in Group I belong to age 10-12 years and majority 35.3% of Group II patients belong to ≤2 years. More than half of the (51.5%) patients had grade III varices followed by 24.2% grade II, 21.2% grade I and only 3.0% had grade IV. All patients had splenomegaly in Group I and 70.6% of Group II, which was significantly (p<0.05) higher in Group I. Jaundice, spider angioma, hepatomegaly and ascites werealmost alike between two groups. Platelet count <1,50,000/mm³ in 72.73% patients of Group I and 41.18% in Group II, which was significantly (p<0.05) higher in Group I. Similarly, Serum albumin <3.5mg/dl in 78.79% patients of Group I and 47.6% of Group II, was also significantly (p<0.05) higher in Group I. Serum bilirubin, serum alanine aminotransferase and International normalized ratio (INR) were almost similar between two groups. Wilson's disease was found in 42.4% of chronic liver disease with esophageal varices and 35.3% of Chronic liver disease without esophageal varices. Hepatitis B virus infections were 6.1% and 5.9% in Group I and Group II respectively. Unknown cause was found 42.4% and 17.6% in Group I and Group II respectively. Splenomegaly, Platelet count <1,50,000/mm³ and serum albumin <3.5mg/dl are important predictors of esophageal varices. These three parameters can be used independently to predict esophageal varices in children with chronic liver disease.

Childhood Wilson Disease: Bangladesh Perspective.

Wilson's disease (WD) is an autosomal recessive disorder affecting copper metabolism causing copper induced damage to various organs. In children liver is commonly involved. Central nervous system, eyes, RBC, kidneys, brain and bones may also be affected. Aim of the study is to evaluate clinical & laboratory profile of Wilson's disease in children. This cross sectional descriptive study was conducted at the department of Paediatric Gastroenterology and Nutrition, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh, from January 2011 to December, 2013. One hundred consecutive children of WD between 3 to 18 years of age were evaluated for clinical & biochemical profile. Mean age of studied children was 8.5±1.5 years. Male female ratio was 2:1. Ninety one percent patients were Muslim and nine percent Hindu. Consanguinity of marriage was found in 30% cases. Seven parents were first degree cousin. Family history of chronic liver disease was present in 15% of patients. Most (53%) cases of the hepatic WD presented between 5 to 10 years of age and most of the neurologic WD manifested in 10-15 years age group. Among 100 patients of WD, 69 children presented only with hepatic manifestations, 6 only with neurological manifestations, 14 with both hepatic & neurological manifestation, 10 children was asymptomatic and 1 patient presented with psychiatric features. WD presented as chronic liver disease (CLD) in 42%, CLD with portal hypertension in 34%, acute hepatitis in 20% and fulminant hepatic failure in 4% cases. Stigmata of chronic liver disease were found in 18% patients. Commonest stigmata was thenar and hypothenar wasting (n=8). Keiser- Fleischser ring (K-F ring) was found in 76% of the total patients. K-F ring was present in 84% ( 58 out of 69) of the hepatic only Wilsonian patients and in 90% (18 out of 20) of all neurologic Wilsonian patients. Asymptomatic and psychiatric patient had no K-F ring. About 26% of the WD patients had Coombs negative hemolytic anemia in PBF. Most of the WD patients had altered liver function. Elevated serum transaminase was found in 85% of all cases, prolonged prothrombin time in 59% cases & low serum albumin in 53% cases. Seventy three percent patients had low serum ceruloplasmin, basal urinary copper of >100µgm/day was found in 81% cases and urinary copper following penicillamine challenge of >1200µgm/day was found in 92% cases. In 28 cases with hepatic presentation esophageal varices were identified by upper gastrointestinal endoscopy. WD patient with hepatic presentations were given zinc sulphate along with penicillamine. All patients with neurological manifestation as well as asymptomatic cases were maintained on zinc therapy. WD is a treatable metabolic cause of liver disease. Majority of studied WD children presented with hepatic manifestation of which 76% presented with CLD. Any child presented with jaundice after the age of 3 years should be investigated for WD.

Klippel Trenaunay Syndrome: A Case Report.

Klippel Trenaunay syndrome refers to a rare congenital anomaly which is characterized by capillary malformation, venous malformation and sometimes lymphatic malformation associated with overgrowth of a limb, with soft tissue hypertrophy and/or bony hypertrophy. The anomaly, if present, is present at birth and usually involves the lower limbs as well as portion of trunk, face, uppper limb or head. Our reporting case is a preterm male neonate having port wine stain, varicose veins and excessive growth of soft tissue of left lower limb clinically consistent with Klippel Trenaunay Syndrome.

Early Outcome of Primary Repair in Colonic Injury.

The management of the colon injury remains controversial in spite of a number of divergent reports during the past decade. Previously surgeons were reluctant to do primary anastomosis but now-a-days they are doing primary repair with good results. The present study is designed to see the early outcomes of primary repair in colonic injury. This prospective observational study performed at Dhaka Medical College Hospital, Dhaka, Bangladesh from January 2013 to June 2013 with allocation of 50 patients with colonic injury who underwent laparotomy with primary repair of that injury in the department of Casualty Surgery. A primary repair was performed after freshening the edges or by resection and primary anastomosis with 3.0 round-body Vicryl by single layer extra mucosal interrupted suture. Data processed using software SPSS version 16.0. For all analytical results a p value <0.05 was considered significant. In this study the commonest site of injury were transvers colon and sigmoid colon 38.0% in each. Out of 50 respondents, 5(10.0%) developed burst abdomen, 1(2.0%) developed entero-cutaneous fistula with none had paralytic ileus or septicaemia or pelvic collection. No mortality observed. This study showed that the increasing in colon injury scale (CIS) score culminate into increasing rate of postoperative complication & post operative complications were more at left colon (24%). On basis of our findings, we recommend the primary repair is a safe and effective surgical technique for addressing the large gut injury. Unnecessary proximal diversions should be avoided. According to our experience, we believe that the policy of primary repair of colon injuries can be applied more liberally in majority of patients with high success rate.

Correlation between Umbilical Cord Diameter and Cross Sectional Area with Gestational Age and Foetal Anthropometric Parameters.

The objective of the study was to find out correlation between umbilical cord diameter, cross sectional area with gestational age and foetal anthropometric parameters. This cross sectional study was conducted among healthy women between the 24(th) and 40(th) completed weeks of a normal pregnancy in the Department of Radiology & Imaging, Mymensingh Medical College Hospital, Mymensingh during the study period, from July 2009 to June 2011. A total of 230 consecutive normal pregnancy patients were included in the study. The diameter & cross-sectional area of the umbilical cord were measured on a plane adjacent to the junction of the umbilical cord and the fetal abdomen, in cross-section, with maximum magnification of the image. The cord was manually circled, and it's cross sectional areas was automatically calculated by the ultrasonograph. The mean±SD age was 24.3±4.7 years with range from 19 to 36 years. The mean gestational age was 32.1±4.5 weeks and more than a half (56.4%) of the pregnant women were nulliparas. A positive significant (p<0.001) correlation were found between umbilical cord diameter with bi-parietal diameter (r=0.548); head circumference (r=0.411); abdominal circumference (r=0.444); femur length (r=0.366) and gestational age gestation age (r=0.643). Similarly, a significant (p<0.001) positive week correlation were found between umbilical cross sectional area with bi-parietal diameter (r=0.3303); head circumference (r=0.3202); abdominal circumference (r=0.2651); femur length (r=0.3307) and gestation age (r=0.4051). A positive significant better correlation was found with umbilical cord diameter than cross sectional area with foetal anthropometric parameters.

Comparison of transvaginal and transabdominal ultrasonography in the diagnosis of ectopic pregnancy.

This cross sectional study was carried out in the department of Radiology and Imaging, Dhaka Medical College Hospital from July 2008 to June 2010 to compare the accuracy of transvaginal ultrasonography and transabdominal ultrasonography in the diagnosis of clinically suspected cases of ectopic pregnancy. Initially 60 patients with clinical suspicion of ectopic pregnancy were included in this study after analyzing selection criteria 30 patients underwent both transvaginal and transabdominal ultrasonography. 'Histopathological diagnosis' was considered gold standard against which accuracies of two diagnostic modalities were compared. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy of transabdominal ultrasonography as a diagnostic modality in evaluation of suspected ectopic pregnancy were 73.1%, 75%, 95%, 30% and 73.3% respectively where as transvaginal ultrasonography was found to have 92.3% sensitivity, 75% specificity, 96% positive predictive value, 60% negative predictive value and 90% accuracy. Transvaginal ultrasonography was superior to transabdominal ultrasonography in the evaluation of suspected ectopic pregnancies so, transvaginal ultrasonography is important for early and accurate diagnosis of ectopic pregnancy.

Age-related changes in triathlon performances.

The aim of this study was two-fold: i) to analyse age-related declines in swimming, cycling, and running performances for Olympic and Ironman triathlons, and ii) to compare age-related changes in these three disciplines between the Olympic and Ironman triathlons. Swimming, cycling, running and total time performances of the top 10 males between 20 and 70 years of age (in 5 years intervals) were analysed for two consecutive world championships (2006 and 2007) for Olympic and Ironman distances. There was a lesser age-related decline in cycling performance (p<0.01) compared with running and swimming after 55 years of age for Olympic distance and after 50 years of age for Ironman distance. With advancing age, the performance decline was less pronounced (p<0.01) for Olympic than for Ironman triathlon in cycling (>55 years) and running (>50 years), respectively. In contrast, an age-related decline in swimming performance seemed independent of triathlon distance. The age-related decline in triathlon performance is specific to the discipline, with cycling showing less declines in performance with age than swimming and running. The magnitude of the declines in cycling and running performance at Ironman distance is greater than at Olympic distance, suggesting that task duration exerts an important influence on the magnitude of the age-associated changes in triathlon performance.

Induction of systemic resistance in Arabidopsis thaliana in response to a culture filtrate from a plant growth-promoting fungus, Phoma sp. GS8-3.

The plant growth-promoting fungus (PGPF), Phoma sp. GS8-3, isolated from a zoysia grass rhizosphere, is capable of protecting cucumber plants against virulent pathogens. This fungus was investigated in terms of the underlying mechanisms and ability to elicit systemic resistance in Arabidopsis thaliana. Root treatment of Arabidopsis plants with a culture filtrate (CF) from Phoma sp. GS8-3 elicited systemic resistance against the bacterial speck pathogen Pseudomonas syringae pv. tomato DC3000 (Pst), with restricted disease development and inhibited pathogen proliferation. Pathway-specific mutant plants, such as jar1 (jasmonic acid insensitive) and ein2 (ethylene insensitive), and transgenic NahG plants (impaired in salicylate signalling) were protected after application of the CF, demonstrating that these pathways are dispensable (at least individually) in CF-mediated resistance. Similarly, NPR1 interference in npr1 mutants had no effect on CF-induced resistance. Gene expression studies revealed that CF treatment stimulated the systemic expression of both the SA-inducible PR-1 and JA/ET-inducible PDF1.2 genes. However, pathogenic challenge to CF-treated plants was associated with potentiated expression of the PR-1 gene and down-regulated expression of the PDF1.2 gene. The observed down-regulation of the PDF1.2 gene in CF-treated plants indicates that there may be cross-talk between SA- and JA/ET-dependent signalling pathways during the pathogenic infection process. In conclusion, our data suggest that CF of Phoma sp. GS8-3 induces resistance in Arabidopsis in a manner where SA and JA/ET may play a role in defence signalling.

Construction of fetal charts for biparietal diameter, fetal abdominal circumference and femur length in Bangladeshi population.

Infants born for small for date (SFD) fetuses have an increased risk of perinatal mortality and morbidity. Different methods have been applied to identify these fetuses including history, clinical examination and ultrasonography. Ultrasonography has a better predictive value and majority of such fetuses can be identified. Measurements of the fetal biparietal diameter (BPD), abdominal circumference (AC) and femur length (FL) charts are widely used in dating pregnancies and follow-up of pregnant women in assessing fetal growth, identification of small for date (SFD) and growth retarded fetuses. This prospective study was performed to construct fetal chart for BPD, AC and FL at different gestational weeks from the Bangladeshi pregnant women. Seven hundred and ten women had ultrasonic measurements of fetal BPD, AC and FL between 12 to 42 weeks of pregnancy. Centiles, mean and the standard deviation (SD) were calculated for BPD, AC and FL. Mean maternal age was 24.73 +/- 4.48 (Mean +/- SD) and 310 (43.7%) were primigravidae. There was a gradual increase of the BPD (outer-inner), AC and FL measurements of 5th, 10th, 50th and 90th Centiles upto 38th weeks of gestation with a gradual increase of SD showing increasing dispersion of data. In cases of BPD and AC, After 38th weeks of gestation the Centiles showed a slower growth rate towards 42 weeks of pregnancy. This slower growth rate from 38 weeks of pregnancy was not noted in case of femur length. Fetal charts with the raw data for each measurement with superimposed fitted lines derived from polynomial (quadratic) regression were constructed. Quadratic model showed good fit to the data during construction of fetal charts. The new fetal measurement charts of BPD, AC and FL are unique for the Bangladeshi population and have not been found similar in the later weeks of pregnancy to those published for other Caucasian populations. These charts will help the clinicians and sonographers in dating pregnancy, identifying SFD and growth retarded fetuses.

Effect of methotrexate and folinic acid on accumulation of iron in mice.

A mouse-model was used to investigate the effect of methotrexate (MTX) and folinic acid on accumulation of iron in young growing mice. Four equal groups of Balb/c young male mice were treated (subcutaneously) with either MTX, or folinic acid, or MTX plus folinic acid, or physiological saline on every second day. After 3 weeks of treatment, liver, spleen, kidney, small intestine, brain, skeletal muscle and heart were removed and analyzed for iron contents using a spectrophotometric method. When the mean values of iron in liver of four groups were compared using one way ANOVA followed by Tukey's HSD test, the group receiving MTX alone was found to have significantly (p=0.004) more accumulation of iron in liver. The group receiving MTX plus folinic acid had iron accumulation in the liver similar to the placebo group. However, the mean values of iron in brain, kidney, small intestine, skeletal muscle, heart and spleen in all the groups, were not found to be statistically different. The data indicate that MTX shifts iron from being utilized in hemoglobin synthesis to liver stores. Folinic acid administration 8 h post-MTX, however, prevents this shift of iron to liver. Decreased levels of iron in plasma in mice treated with MTX alone suggest decreased availability of iron to other tissues for their normal growth and development.

The peri-albumen layer: a novel structure in the envelopes of an avian egg.

The present paper describes a novel structure, termed the peri-albumen layer, in the egg-envelopes of the quail Coturnix japonica. It reacts with Alcian blue and exists between the egg white and the shell membrane. Ultrastructurally, it is of fine granular structure and forms a fenestrate sheet, the width of which is 190 nm or less. Isolated materials of the peri-albumen layer include an Alcian-blue-positive polysaccharide of 260 kDa, and three glycoproteins of 160, 108 and 52 kDa. The layer is supplied to an egg when it passes through the magnum-isthmus junction, the normalized length of which is 0.62-0.63 of the oviduct. The mucosa of the junction consists exclusively of a luminal epithelium. It is apparently distinct from the mucosa of the magnum and the isthmus, which consist of a luminal epithelium and tubular glands. The luminal epithelium of the magnum-isthmus junction stains prominently with Alcian blue and consists of alternately distributed ciliated cells and granular cells. Immunohistochemistry with an antiserum raised against the materials of the peri-albumen layer revealed the staining of the peri-albumen layer of the egg, and secretory cells of the luminal epithelium at the magnum-isthmus junction. It was concluded that the materials of the peri-albumen layer are produced by secretory cells at the magnum-isthmus junction of the oviduct.

Folinic acid protects against suppression of growth by methotrexate in mice.

The objective of this study was to investigate whether folinic acid supplementation would protect young mice against suppression of growth by methotrexate (MTX). Four equal groups of Balb/c young male mice (5 animals in each group; mean+/-SD body weight 9.64+/-0.85 g, in their rapid growth phase) were subjected to the following drug treatment: One group was given MTX (3.5 mg/kg body weight) intraperitoneally on every 2nd day, another received folinic acid (7.0 mg/kg body weight) intraperitoneally every 2nd day. The third group was given both of these drugs (MTX on every 2nd day and folinic acid 8 h post-MTX injection). The fourth group was injected with physiological saline every other day to serve as a control group. Total body weight, food and water consumption by animals in each group were monitored every second day for a period of 3 weeks. After this period mice were sacrificed and liver, spleen and kidneys were excised, weighed and analyzed for MTX and dihydrofolate reductase activity. A small segment of the proximal part of small intestine and small pieces of liver and kidney were also removed to study morphological changes. Compared to the groups, which received folinic acid alone, folinic acid plus MTX or physiological saline, mean increase in body weight (6.8+/-0.8 g) of mice over a period of 3 weeks was minimal in the group receiving MTX alone (one-way ANOVA p=0.0001). The mean weights of liver and kidney in this group receiving MTX alone were also found to be significantly less than the mean weights of these organs in the 3 groups (p<0.001). The negative effect on growth of animals appears not only due to malabsorption but inhibition of pathway of de novo DNA synthesis may also be involved. This is supported by loss of villous pattern in small intestine of mice treated with MTX alone and increased accumulation of free MTX and decreased dihydrofolate reductase in the liver of the group receiving MTX alone as compared with the group receiving MTX plus folinic acid. The data indicate that the administration of folinic acid protects mice against suppression of growth by MTX. On the basis of these observations it can be deduced that patients suffering from juvenile rheumatoid arthritis or acute lymphoblastic leukaemia receiving MTX over a long period of time might be at a risk of experiencing short-term suppression of growth, however they could benefit from supplementation with folinic acid.

Induction of white cell proliferation due to haematopoietic growth factors is associated with an increase in multiple forms of dihydrofolate reductase in non-neutropenic cancer patients.

OBJECTIVE: Granulocyte-colony stimulating factor (G-CSF) and granulocyte macrophage-colony stimulating factor (GM-CSF) are frequently used in cancer patients to overcome the granulocytopenic effects of chemotherapy, and also to mobilize the stem cells. The mobilized stem cells are collected from the peripheral blood and used for transplantation following high doses of chemotherapy. However, the molecular mechanism by which these colony stimulating factors (CSFs) bring about proliferation of myeloid precursor cells is not clearly known. Dihydrofolate reductase (DHFR), which has an established role in DNA synthesis, could be a link between administration of CSF and stem cell proliferation. The purpose of this study was to investigate whether CSFs induce white cell proliferation by producing multiple forms of DHFR. METHODS: Twelve patients with non-haematological malignancies were treated with either G-CSF or GM-CSF to mobilize stem cells. Nine healthy subjects were treated with placebo as controls. Blood samples were obtained before and after stimulation with CSFs or placebo. White blood cells were separated and concentrations of both active DHFR and immunoreactive nonfunctional form of DHFR were determined in their cytoplasm using methotrexate-binding assay and enzyme-linked immunosorbent assay, respectively. Total leucocytes count (TLC) was also monitored before and after stimulation with CSFs or placebo. RESULTS: There was a significant (P < 0.05) increase in concentration of immunoreactive nonfunctional form of DHFR and TLC following stimulation with CSFs. There was an increase in concentration of active DHFR as well, however, this did not reach statistical significance. In the placebo-treated subjects, no significant increase in active DHFR, immunoreactive nonfunctional form of enzyme or TLC was observed. However, it was noticed that the base-line values of active DHFR and immunoreactive nonfunctional form of enzyme in leucocytes of cancer patients were higher than the base-line values in leukocytes of normal healthy subjects. CONCLUSION: Our data suggest that colony stimulating factors induce white cell proliferation by increasing levels of multiple forms of DHFR.

Increased levels of multiple forms of dihydrofolate reductase in peripheral blood leucocytes of cancer patients receiving haematopoietic colony-stimulating factors: interim analysis.

The precise mechanism whereby granulocytes proliferate when haematopoietic colony stimulating factors (CSFs) are used in neutropenic cancer patients is poorly understood. The purpose of this study was to investigate whether these cytokines bring about leucocyte proliferation by increasing the levels of multiple forms of dihydrofolate reductase (DHFR). Blood samples were collected from 36 cancer patients (25 males and 11 females) with chemotherapy-induced neutropenia. One sample of blood from each patient was obtained before therapy either with CSF, such as granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) or with placebo, and another one at the time of resolution of neutropenia. Peripheral blood leucocytes in these blood samples were counted, separated and lysed. From lysates, cytoplasmic samples were prepared and analyzed for active DHFR by a methotrexate-binding assay and for total immunoreactive DHFR by an enzyme linked immunosorbent assay. The increase in total leucocyte count (TLC) was most prominent (P < 0.005) in the CSF group and less so (P < 0.05) in the placebo group. The mean +/- SD concentration values of active DHFR before and after stimulation with GM-CSF found were to be 0.34 +/- 0.4 ng/mg protein and 0.99 +/- 0.82 ng/mg protein, respectively, and in the group treated with G-CSF, 0.24 +/- 0.32 ng/mg protein and 1.18 +/- 2.4 ng/mg protein, respectively. This increase in active DHFR after stimulation with CSF was statistically significant (P < 0.05). Similarly, concentration values of immunoreactive but nonfunctional form of DHFR (IRE) were 110 +/- 97 ng/mg protein and 605 +/- 475 ng/mg protein before and after stimulation with GM-CSF, and 115 +/- 165 ng/mg protein and 1,054 +/- 1,095 ng/ mg protein before and after stimulation with G-CSF. This increase in concentration of IRE after stimulation with GM-CSF or G-CSF was statistically significant (P < 0.005). In the control group, there was an increase in the concentration of both active DHFR and IRE after treatment with placebo. However, this was not statistically significant. Resolution of neutropenia was quicker in the groups treated with CSF compared to the control group. Results of this study indicate that colony stimulating factors (G-CSF and GM-CSF) induce white cell proliferation by increasing the levels of multiple forms of DHFR.

Description of Arthrobacter creatinolyticus sp. nov., isolated from human urine.

Three strains of creatinine-hydrolysing bacteria isolated from human urine were characterized taxonomically. They were aerobic, non-spore-forming, Gram-positive rods with the peptidoglycan of the cell wall containing lysine. MK-8 and MK-9 were found to be the major types of menaquinone. The G + C content of the DNA was 66-67 mol%. The 16S rRNA sequence of one strain (GIFU 12498) was determined and aligned with other high-G + C-content Gram-positive rods from different genera. Following phylogenetic analysis, this strain was placed in the genus Arthrobacter. Arthrobacter protophormiae was the most closely related species in the phylogenetic tree, and this species also showed the highest sequence homology value (97%) with GIFU 12498. However, DNA-DNA hybridization indicated that GIFU 12498 did not belong to A. protophormiae (33.8 +/- 3.5% chromosomal similarity). The three urine strains belonged to one species because they shared more than 95% DNA-DNA similarity. It is proposed that these strains are placed in the genus Arthrobacter as a new species, Arthrobacter creatinolyticus sp. nov. The type strain of A. creatinolyticus is GIFU 12498, which has been deposited in the Japan Collection of Microorganisms (JCM) with the accession number JCM 10102.