A priori estimation of accuracy and of the number of wells to be employed in limiting dilution assays

The use of limiting dilution assay (LDA) for assessing the frequency of responders in a cell population is a method extensively used by immunologists. A series of studies addressing the statistical method of choice in an LDA have been published. However, none of these studies has addressed the point of how many wells should be employed in a given assay. The objective of this study was to demonstrate how a researcher can predict the number of wells that should be employed in order to obtain results with a given accuracy, and, therefore, to help in choosing a better experimental design to fulfill one's expectations. We present the rationale underlying the expected relative error computation based on simple binomial distributions. A series of simulated in machina experiments were performed to test the validity of the a priori computation of expected errors, thus confirming the predictions. The step-by-step procedure of the relative error estimation is given. We also discuss the constraints under which an LDA must be performed.

Plasma and testicular testosterone levels, volume density and number of Leydig cells and spermatogenic efficiency of rabbits

Plasma and tissue testosterone concentrations were determined by radioimmunoassay in 12 eight-month-old sexually mature New Zealand White rabbits and evaluated for possible associations with spermatogenic efficiency as well as with volume density and number of Leydig cells. Testicular tissue was processed histologically and histometry was performed in order to quantify germ cells, Sertoli cells and Leydig cells. Spermatogenic efficiency, reported as the ratios among germ cells (spermatogonia, primary spermatocytes and round spermatids) and by the ratio of germ cells to Sertoli cells, was not associated with testosterone levels. However, Leydig cell parameters such as number of Leydig cells per gram of testis, total number of Leydig cells per testis and percent cell volume of Leydig cell nuclei were correlated significantly with testosterone levels. The statistically significant correlation (r = 0.82, P<0.05) observed between testosterone levels and the number of Leydig cells per gram of testis suggests that, in the rabbit, the latter parameter can serve as a criterion for monitoring testosterone levels in this species under normal conditions.

Trypanosoma cruzi infection and/or administration of the nonsteroidal anti-inflammatory nimesulide increase the number of colonic crypts overexpressing metallothioneins in rat colon carcinogenesis

Trypanosoma cruzi infection and nonsteroidal anti-inflammatory drugs inhibit colorectal carcinogenesis by mechanisms not completely known and metallothionein proteins (MTs) may be involved in this process. Sixty-six male Wistar rats weighing 90 to 120 g were randomly divided into seven groups (GI to GVII). GI, GII and GIII animals were subcutaneously infected with 200,000 trypomastigote forms of the Y strain of T. cruzi. After 8 weeks, GI, GII, GIV, and GVI were injected with one weekly subcutaneous dose of 12 mg/kg dimethylhydrazine for 4 weeks. In sequence, GI, GIV and GV were treated with nimesulide (10 mg/kg per dose, five times per week for 8 weeks). Groups I, III, IV, and VI had 12 animals, and each of the other groups had 6 animals. All the animals were euthanized 8 weeks after the last dimethylhydrazine injection. The colons were fixed and processed for MT immunohistochemistry. The index of MT-overexpressing colonic crypts (MTEC) was estimated as the percentage of MT-stained crypts in relation to the total number of crypts scored. Five hundred crypts per animal were scored. Data were analyzed by the Kruskal-Wallis test followed by the Dunn test. There was an increase in MTEC index in the groups either infected with T. cruzi or treated with nimesulide or both infected and treated when compared to control (401, 809, and 1011%, respectively). We suggest that the increased formation of MTEC may be related to the protection against carcinogenesis provided both by T. cruzi infection and nimesulide.

Long-term aerobic swimming training by rats reduces the number of aberrant crypt foci in 1,2-dimethylhydrazine-induced colon cancer

We determined the effect of long-term aerobic swimming training regimens of different intensities on colonic carcinogenesis in rats. Male Wistar rats (11 weeks old) were given 4 subcutaneous injections (40 mg/kg body weight each) of 1,2-dimethyl-hydrazine (DMH, dissolved in 0.9% NaCl containing 1.5% EDTA, pH 6.5), at 3-day intervals and divided into three exercise groups that swam with 0% body weight (EG1, N = 11), 2% body weight (EG2, N = 11), and 4% body weight of load (EG3, N = 10), 20 min/day, 5 days/week for 35 weeks, and one sedentary control group (CG, N = 10). At sacrifice, the colon was removed and counted for tumors and aberrant crypt foci. Tumor size was measured and intra-abdominal fat was weighed. The mean number of aberrant crypt foci was reduced only for EG2 compared to CG (26.21 ± 2.99 vs 36.40 ± 1.53 crypts; P < 0.05). Tumor incidence was not significantly different among groups (CG: 90%; EG1: 72.7%; EG2: 90%; EG3: 80%). Swimming training did not affect either tumor multiplicity (CG: 2.30 ± 0.58; EG1: 2.09 ± 0.44; EG2: 1.27 ± 0.19; EG3: 1.50 ± 0.48 tumors) or size (CG: 1.78 ± 0.24; EG1: 1.81 ± 0.14; EG2: 1.55 ± 0.21; EG3: 2.17 ± 0.22 cm³). Intra-abdominal fat was not significantly different among groups (CG: 10.54 ± 2.73; EG1: 6.12 ± 1.15; EG2: 7.85 ± 1.24; EG3: 5.11 ± 0.74 g). Aerobic swimming training with 2% body weight of load protected against the DMH-induced preneoplastic colon lesions, but not against tumor development in the rat.

Fish oil ingestion reduces the number of aberrant crypt foci and adenoma in 1,2-dimethylhydrazine-induced colon cancer in rats

We determined the effect of fish oil (FO) ingestion on colonic carcinogenesis in rats. Male Wistar rats received 4 subcutaneous injections (40 mg/kg body weight each) of 1,2-dimethylhydrazine (DMH) at 3-day intervals and were fed a diet containing 18% by weight FO (N = 10) or soybean oil (SO, N = 10) for 36 weeks. At sacrifice, the colon was removed, aberrant crypt foci were counted and the fatty acid profile was determined. Intestinal tumors were removed and classified as adenoma or carcinoma. Liver and feces were collected and analyzed for fatty acid profile. FO reduced the mean (± SEM) number of aberrant crypt foci compared to SO (113.55 ± 6.97 vs 214.60 ± 18.61; P < 0.05) and the incidence of adenoma (FO: 20% vs SO: 100%), but carcinoma occurred equally in FO and SO rats (2 animals per group). The polyunsaturated fatty acid (PUFA) profile of the colon was affected by diet (P < 0.05): total ω-3 (FO: 8.18 ± 0.97 vs SO: 1.71 ± 0.54%) and total ω-6 (FO: 3.83 ± 0.59 vs SO: 10.43 ± 1.28%). The same occurred in the liver (P < 0.05): total ω-3 (FO: 34.41 ± 2.6 vs SO: 6.46 ± 0.59%) and total ω-6 (FO: 8.73 ± 1.37 vs SO: 42.12 ± 2.33%). The PUFA profile of the feces and liver polyamine levels did not differ between groups (P > 0.05). In conclusion, our findings indicate that chronic FO ingestion protected against the DMH-induced preneoplastic colon lesions and adenoma development, but not against carcinoma in rats.

Impact of a 2-year intervention program on cardiometabolic profile according to the number of goals achieved

We investigated the impact of lifestyle goal achievement on cardiovascular risk factors after a 2-year behavioral intervention program applied to 394 adults (113 with diabetes, mean age 60.2 ± 11.4 years, 56% women) and targeting four goals: ≥5% weight loss; ≥150 min/week physical activities; <10% saturated fat intake/day; ≥400 g fruit and vegetable intake/day. Baseline characteristics and changes in variables after intervention among the four categories of number of goals achieved (none, 1, 2, and ≥3) were compared by independent ANOVA or the Kruskal-Wallis test. Individuals without diabetes achieving a higher number of goals were more likely to be older (3 or 4 goals: 61.8 ± 12.6 years vs none: 53.3 ± 10.3 years, P < 0.05) and to have a lower mean BMI (3 or 4 goals: 21.7 ± 2.6 kg/m² vs none: 29.0 ± 4.8 kg/m², P < 0.05), diastolic blood pressure (3 or 4 goals: 77.3 ± 2.1 mmHg vs none: 85.4 ± 9.6 mmHg, P < 0.05), triglyceride (3 or 4 goals: 116.1 ± 95.1 mg/dL vs none: 144.8 ± 65.5 mg/dL, P < 0.05) and insulin levels (3 or 4 goals: 3.6 ± 2.4 μU/L vs none: 5.7 ± 4.0 μU/L, P < 0.05) than those achieving fewer goals. The absolute changes in cardiovascular risk factors tended to be more pronounced with increasing number of goals achieved in individuals without diabetes. The intervention had a beneficial impact on the cardiometabolic profile of individuals with normal or altered glucose metabolism. The number of goals achieved in this lifestyle intervention was associated with the magnitude of improvement of cardiovascular risk factors in individuals without diabetes. Participants with a better cardiometabolic profile seemed to be more likely to have a healthy lifestyle.

Serum levels of brain-derived neurotrophicfactor correlate with the number of T2 MRI lesions in multiple sclerosis

The objective of the present study was to determine if there is a relationship between serum levels of brain-derived neurotrophic factor (BDNF) and the number of T2/fluid-attenuated inversion recovery (T2/FLAIR) lesions in multiple sclerosis (MS). The use of magnetic resonance imaging (MRI) has revolutionized the study of MS. However, MRI has limitations and the use of other biomarkers such as BDNF may be useful for the clinical assessment and the study of the disease. Serum was obtained from 28 MS patients, 18-50 years old (median 38), 21 women, 0.5-10 years (median 5) of disease duration, EDSS 1-4 (median 1.5) and 28 healthy controls, 19-49 years old (median 33), 19 women. BDNF levels were measured by ELISA. T1, T2/FLAIR and gadolinium-enhanced lesions were measured by a trained radiologist. BDNF was reduced in MS patients (median [range] pg/mL; 1160 [352.6-2640]) compared to healthy controls (1640 [632.4-4268]; P = 0.03, Mann-Whitney test) and was negatively correlated (Spearman correlation test, r = -0.41; P = 0.02) with T2/FLAIR (11-81 lesions, median 42). We found that serum BDNF levels were inversely correlated with the number of T2/FLAIR lesions in patients with MS. BDNF may be a promising biomarker of MS.

Letters from the Secretary of War to the Committee of Ways and Means, in relation to the number of Militia called into the public service in 1813, to a provision for paying the bounties and premiums to soldiers lately authorized, and to the strength of the army.--

At head of title: [78].

Persons who served more than six months in the War of 1812 : letter from the Secretary of War, transmitting in compliance with a resolution of the House calling for a statement of the number of officers, non-commissioned officers, privates, &c., who served for a period of six months and upwards in the War of 1812.--

Caption title.

The Number of Militia Called into the Public Service in 1813

Full Title: Letters from the Secretary of War to the Committee of Ways and Means, in relation to the number of Militia called into the public service in 1813, to a provision for paying the bounties and premiums to soldiers lately authorized, and to the strength of the army March, 3, 1814. Read, and ordered to be printed. U.S. 13th Congress 2nd Session, 1813-1814. House.

List of the number of loads dredged by Smiley’s Dredge since the 1st of October along the Welland Railway

List of the number of loads dredged by Smiley’s Dredge since the 1st of October along the Welland Railway. This is addressed to S.D. Woodruff and signed by James Woodall of Lock No. 1. There are holes and stains in the document. Text is not affected, Jan. 12, 1859.

Note regarding the number of days Fred Holmes was employed upon the Port Robinson and Thorold macadamized road during the months of July and August

Note regarding the number of days Fred Holmes was employed upon the Port Robinson and Thorold macadamized road during the months of July and August. This is signed by S.D. Woodruff and Fred Holmes, November, 1857.

Median Sets and Median Number of a Graph

A profile is a finite sequence of vertices of a graph. The set of all vertices of the graph which minimises the sum of the distances to the vertices of the profile is the median of the profile. Any subset of the vertex set such that it is the median of some profile is called a median set. The number of median sets of a graph is defined to be the median number of the graph. In this paper, we identify the median sets of various classes of graphs such as Kp − e, Kp,q forP > 2, and wheel graph and so forth. The median numbers of these graphs and hypercubes are found out, and an upper bound for the median number of even cycles is established.We also express the median number of a product graph in terms of the median number of their factors.