Evaluation of the permeability of hair growing ingredient encapsulated PLGA nanospheres to hair follicles and their hair growing effects

This paper describes the process of encapsulating hair growing ingredients in the PLGA nanospheres by emulsion solvent diffusion method and investigates the feasibility of using the PLGA nanospheres as the DDS (Drug delivery System) carriers for delivering various hair growing ingredients to hair follicles. In-vitro and in-vivo tests were conducted to verify the performances of encapsulated PLGA nanospheres with three different hair growing ingredients. In the in-vitro tests, the scalp-pore permeability of hair growing ingredient encapsulated PLGA nanospheres (dispersed in the PBS solution) was examined using human scalp biopsies in a modified Bronaugh diffusion chamber in comparison to that of the control samples containing the hair growing ingredient in the PBS solution. Furthermore, the hair growing effect of the encapsulated PLGA nanospheres was evaluated with the C3H mice in the in-vivo tests. By observing the fluorescence intensity of the ingredients, as shown in the cross-section photographs of the human scalp biopsies, it was found that the dispersion liquids containing hair growing ingredient encapsulated PLGA nanospheres exerted a scalp-pore permeability 2.0- to 2.5-fold more marked than that of the control samples. Also, the hair growing activities were enhanced by using the encapsulated PLGA nanospheres, which transformed the hair growth cycle from the resting phase to the growing phase. As a result, the degree of hair growth was improved significantly. These results suggested that the PLGA nanosphere can be a new DDS carrier for delivering hair growing ingredients and drugs to the hair follicles.     

Changes in trace element concentrations in hair of growing children

Head hair concentrations of zinc, copper, manganese, and iron from a total of 418 subjects (154 male and 264 female) aged between 6 mo and 20 yr were measured mainly with flameless atomic absorption spectrophotometry. Only zinc analysis of a part of the female samples (n=140) were analyzed with inductively coupled plasma-atomic emission spectrometry. The two analytical methods showed close agreement. The mean concentration of copper and manganese were significantly higher in male subjects than in female subjects. The trace element concentrations in hair varied with the subject’s age. Zinc concentration in hair decreased from 6 mo to 14 yr in the male subjects and decreased from 6 mo to 12 yr in the female subjects. Then, the concentrations increased gradually to 20 yr in the both sexes. Age-dependent variations of copper and manganese concentrations in hair showed similar trends to those of zinc. The results of this study suggest that a higher concentration in the diet of these trace elements may be required for growing children, especially in the period of adolescence.     

Initial kinetics of Protein turnover in growing yeast

Proteins in yeast growing in a medium with glucose or ethanol as carbon source were pulse-labelled by a 20-min incubation with¹⁴C-leucine. The proteins in cells labelled and growing in a glucose medium were stable; when this population was transferred to the ethanol medium, the proteins were degraded at a rate of 1.1 %/h. The population labelled and growing in an ethanol medium displayed a fraction of short-lived proteins (about 4 %), decaying with a half-life of 0.5 h. The size of the short-lived protein fraction increased slightly after shifts to a glucose as well as to a starvation medium. The residual long-lived proteins underwent a turnover of 1.3 –1.4 %/h in the ethanol or the starvation medium and of 0.3 %/h in the glucose medium, respectively. Proteins labelled in the presence of canavanine or ethionine were degraded at only a slightly greater rate than the normal proteins. Participant of the UNESCO Postgraduate Course “On Modern Problems in Biology”.     

Structural study of spongiosa tissue in growing sheep.

A study was made of 160 long bones taken from 40 native Merino sheep of both sexes. These animals, which represented uniform growth (mean growth-curve values), were divided into four groups which were slaughtered consecutively at 0, 45, 105 and 270 days old (0, 6.4, 15 and 38.5 weeks, respectively). The following bones were studied; humerus, femur, tibia and os coxae. Thin lamellae taken from the metaphyses of the bones obtained were fixed, decalcified and stained with hematoxylin-eosin to assess the development of the various components of growing bony tissue. The bones studied followed the same maturation pattern; the os coxae proved to be the best histological indicator in differentiating the age of the animals studied.     

Weight adjustment equation for hair sheep raised in warm conditions

To estimate the nutritional requirements of hair sheep, knowledge about the animal’s weight and its relationships with growth performances is essential. A study was carried with the objective to establish the relationships between BW, fasting BW (FBW), empty BW (EBW), average daily gain (ADG) and empty BW gain (EBWG) for hair sheep in growing and finishing phases in Brazilian conditions. Databases were obtained from 32 studies, for a total of 1145 observations; there were 3 sex classes (non-castrated male, castrated male and female) and 2 feeding systems (pasture and feedlot). The most representative breeds in the database were Santa Ines (n = 473), Morada Nova (n = 70) and Brazilian Somali (n = 47). The other animals in the database were crossbreeds (n = 555). The FBW (kg), EBW and EBWG (kg/day) were estimated according to linear regression. A random coefficient model was adopted, considering the study as a random effect and including the possibility of covariance between the slope and the intercept. The coefficients obtained from the linear regression of the FBW against the BW, EBW against the FBW and EBWG against the ADG did not differ between sex class (P > 0.05) and genotype (P > 0.05). The equations generated to estimate FBW from the BW, EBW from the FBW and EBWG from the ADG are as follows: FBW = −0.5470 (±0.2025) + 0.9313(±0.019) × BW, EBW = −1.4944 (±0.3639) + 0.8816 (±0.018) × FBW and EBWG = 0.906 (±0.019) × ADG, respectively. The low mean squared error values found in the cross-validation confirmed the reliability of these equations. Considering a sheep with a BW of 30 kg and a 100 g ADG, the estimated FBW, EBW and EBWG calculated using the generated equations are 27, 22.65 and 0.090 kg, respectively. In conclusion, the generated equations can be used in growing hair sheep. The validation procedure applied to the generated equations showed that its use for hair sheep seems to be appropriate.     

Gene expression in sheep skin and wool (hair)

We sequenced 2939 ESTs from fetal and adult sheep skin. Stages of gestation were picked to coincide with the major events in skin appendage (wool follicle) formation. Clustering analysis generated a nonredundant set of ESTs 2435 strong (83% nonredundant). Approximately 24% of these gave no hit to NCBI build 29 of the human genome, while 35% were tentatively classified by putative function based on BLASTX hits with a p(N) of <10(-4). In addition to bioinformatics analysis of our ESTs and gene mapping, we have generated a large EST spatial expression data set using in situ hybridization. One thousand one hundred forty-two ESTs have been used for in situ localization; about 31% are from adult sheep skin, 39% from late gestation fetal sheep skin, and 30% from midgestation fetal sheep skin. These probes have been used in over 3000 hybridization experiments. In this report, we summarize the results of in situs on adult sheep skin.     

Phosphorus uptake kinetics in a biofilm sequencing batch reactor

Using data from previously reported experimental work, theoretical kinetic analysis of the aerobic uptake of phosphate is presented. The data obtained in biofilm sequencing batch laboratory reactor was adjusted with polynomial regression to fit a curve during the uptake phase and then analyzed with two kinetic models: Classical Michaelis-Menten and one considering the enzymes involved have allosteric character. Data from nine experimental runs, every one under different operational conditions, were analyzed. The results show that the experimental values used fit in the model of an allosteric enzyme or enzyme-complex with four-binding sites. Based on this consideration, a catalytic model of sequential interactions was obtained which allows the interpretation of the kinetic mechanisms involved in the process. The calculated phosphorus uptake rate values do not show significant differences from the experimental ones.     

Tropical tannin-rich fodder intake modifies saliva-binding capacity in growing sheep

We evaluated the effect of feeding dietary tannins from Lysiloma latisiliquum fresh forage on the saliva tannin-binding capacity of hair sheep lambs without previous exposure to tannin-rich (TR) fodder. Twenty-four hair sheep lambs (13.6±3.04 kg LW) were fed a tannin-free diet at the beginning of the experimental period (from day 10 to 13). On day 14, lambs were distributed into three groups (n=8): control group (CG), fed with the tannin-free diet (from D10 to D112); tannin short-term group (TST), fed the basal diet and 650 g of L. latisiliquum forage (from D14 to D55); tannin long-term group (TLT), fed the basal diet and 650 g of L. latisiliquum forage (from D14 to D112). Saliva samples were collected from the mouth of each lamb in the morning before feeding time on D10 and D14 (baseline period), on D49 and D56 (period 1) and on D97 and D112 (period 2). The tannin binding response of salivary protein (∆% turbidity) was determined with the haze development test (HDT) using either tannic acid or L. latisiliquum forage acetone extract. A turbidity protein index (TPI) was calculated as (∆% turbidity/[salivary protein (mg)]). Differences in HDT and TPI in the different groups were compared by repeated measures ANOVA using Proc Mixed. All groups had similar ∆% turbidity throughout the experiment (P>0.05). At baseline and period 1, the TPI of the different groups was similar (P>0.05). On period 2 the TLT group showed higher TPI compared with CG (P<0.05). Meanwhile, CG and TST showed similar salivary TPI. The saliva of hair sheep lambs consuming TR L. latisiliquum fresh fodder (TLT group) increased their TPI compared with control lambs not exposed to tannins.     

The kinetics of specific serum antibodies in sheep infested with sheep bot larvae]

The kinetics of specific antibodies of the blood serum of sheep experimentally infested with 80, 160 and 1000 specimens of Oestrus ovis larvae was examined. The affinity pure serum IgG and the immunoferment analysis (ELISA) were used for qualitative estimation of specific antibodies. It has been shown that the level of specific antibodies correlates with the larval biomass and is connected with ontogenesis of this parasite. The younger animals, which were infested for the first time, are characterized by more intensive production of specific IgG than adult reinfested ones. The ways of immunity response formation in animals infested with Oestrus ovis larvae are considered.     

Mitochondrial analysis sheds light on the origin of hair sheep

A total of 180 mtDNA sequences from hair Caribbean (93), West African (73) and Canarian‐wooled (14) sheep were analysed to shed light on the origin of hair sheep. A comparison of 360 Iberian sheep sequences retrieved from GenBank was performed to assess a possible European origin of the Caribbean hair sheep. These 180 sequences gave 48 different haplotypes (16 in Caribbean sheep). All Caribbean and Canarian‐wooled sequences and 91.8% of the West African samples belonged to haplogroup B. The sheep analysed showed wide haplotypic identity. Caribbean sheep shared roughly two‐thirds of their samples with Canarian‐wooled and West African samples, respectively. Principal component analysis showed that the Caribbean and the Canarian‐wooled sheep clustered together. Additional analyses showed that hair and Iberian sheep had wide genetic identity. It was not possible to ascertain a single Canarian, African or European origin of the Caribbean hair sheep using mtDNA markers only. European, African and Caribbean hair sheep maternal genetic backgrounds likely result from related domestication events.     

Growth performance and carcass traits of forage-fed hair sheep wethers

The objective of the present study was to compare live animal performance and carcass characteristics of 3/4 or 7/8 Dorper (DO; n = 30), purebred Katahdin (KA; n = 20), purebred St. Croix (SC; n = 17) and purebred Suffolk (SU; n = 10) lambs born in the spring and fall of 2001. After weaning, lambs were supplemented with up to 680 g of a corn-soybean meal supplement while grazing bermudagrass pastures overseeded with ryegrass. Lambs were slaughtered at approximately 210 d of age. From birth to weaning, DO lambs gained faster (P < 0.001) than KA or SC lambs, whereas KA lambs had higher (P < 0.001) ADG than SC lambs. Additionally, DO and SU wethers had greater (P < 0.02) ADG from weaning to slaughter than SC or KA wethers. Suffolk lambs were heavier (P < 0.001) at slaughter and produced heavier (P < 0.001) carcasses than lambs from hair-sheep breeds. Carcasses of KA lambs were fatter (actual fat thickness; P < 0.02) resulting in higher yield grades (P < 0.03) than carcasses of DO, SC, or SU lambs. Carcasses of DO and SU had larger (P < 0.001) longissimus muscle (LM) areas than those of KA or SC carcasses, whereas kidney fat weight and percentage were greater (P < 0.001) in carcasses from KA and SC than DO and SU lambs. Lean maturity was similar (P = 0.32) among breed-types. However, skeletal maturity was greater (P < 0.001) in SU than hair-sheep carcasses. Flank-streaking scores were similar (P = 0.19) among the breed-types, but conformation scores were higher (P < 0.001) for DO and SU carcasses and resulted in higher (P < 0.001) quality grades than SC carcasses. The LM of SU lambs was lighter (higher L^* values; P < 0.05) than that of KA and SC lambs, whereas the LM from DO lambs was redder (higher a^* values; P < 0.001) than SC and SU and more (P < 0.001) yellow than that of the other breed-types. Chops from SU lambs were tougher (higher shear force values; P < 0.007) than chops from the hair-sheep breeds. Results of this study indicate that ADG, carcass muscularity and meat quality were similar between DO and SU lambs, and, although fatter, carcass muscularity of KA was similar to that of DO lambs.     

Estrus synchronization and artificial insemination of hair sheep ewes in the tropics

Hair sheep ewes (St. Croix White and Barbados Blackbelly) were used to evaluate 3 methods of estrus synchronization for use with transcervical artificial insemination (TAI). To synchronize estrus, ewes (n = 18) were treated with PGF2alpha (15 mg, im) 10 d apart, with controlled internal drug release (CIDR) devices containing 300 mg progesterone for 12 d (n = 18), or with intravaginal sponges containing 500 mg progesterone for 12 d (n = 18). On the day of the second PGF2alpha injection or at CIDR or sponge removal, sterile rams were placed with the ewes. Jugular blood samples were collected from the ewes at 6-h intervals until the time of ovulation, and daily for 16 d after estrus (Day 0). Plasma was harvested and stored at -20 degrees C until LH, and progesterone concentrations were determined by RIA. There was no difference (P>0.10) in time to estrus among the CIDR-, PGF2alpha- or sponge-treated ewes. All of the ewes in the CIDR group and 94.4% of the sponge treated ewes exhibited estrus by 36 h after ram introduction, while only 72.2% of PGF2alpha-treated ewes showed signs of estrus by this time (P<0.06). The time from ram introduction to ovulation was not different (P>0.10) among the CIDR-, PGF2alpha- or sponge-treated ewes. The time to the preovulatory LH surge was similar (P>0.10) among CIDR, PGF2alpha and sponge treated ewes. Progesterone levels through Day 16 after the synchronized estrus were not different (P>0.10) among treatment groups. Hair sheep ewes (n = 23) were synchronized using PGF2alpha and bred by TAI using frozen-thawed semen 48 h after the second injection. The conception rate to TAI was 2/23 (8.7%) and produced 3 ram lambs. In a subsequent trial, 17 ewes were synchronized with CIDR devices and bred by TAI using frozen-thawed semen 48 h after CIDR removal, resulting in a conception rate of 52.9% (9/17). It is possible to synchronize estrus in hair sheep using either CIDRs, sponges or PGF2alpha. Even though there were no significant differences in the timing of ovulation or the LH surge among the treatment groups, a higher conception rate was achieved in ewes synchronized with CIDR devices during the second trial. This may reflect an increase in the skill level of the TAI technician.     

Induction kinetics of RNA and proteins in exponentially growing organisms.

A mathematical model of the induction kinetics of RNAs and proteins in exponentially growing organisms is derived, and the cellular concentrations of the induced macromolecules at a given time after induction are related to three parameters: the fraction of the synthesis of these macromolecules in total synthesis, the half life of the inducible macromolecules, and the generation time of the organisms. The model predicts that the concentrations of the inducible macromolecules reach one half of the maximum induction level within one generation time after the onset of the induction. The model also predicts that induction curves of proteins are parabolic when their mRNAs are short-lived, but sigmoid when they are stable. Observed induction curves of beta-galactosidase in Escherichia coli cells fit in the theoretical induction curves.     

Pulmonary neutrophil kinetics in sheep: effects of altered hemodynamics

We investigated the effect of elevated left atrial pressure and reduced cardiac output on pulmonary neutrophil kinetics in the sheep. Sheep neutrophils were isolated, labeled with 111In-oxine, and reinfused. Erythrocytes were labeled with [99mTc]pertechnetate. A gamma camera measured the lung activities of the labeled neutrophils and erythrocytes. The results indicated that 38.5% of the total injected neutrophils marginated in the lung. Pulmonary hemodynamics were altered by inflating a left atrial balloon three times in each sheep for 15-30 min to achieve 5- to 25-mmHg increments in pulmonary arterial wedge pressure. At least a 30-min recovery period was allowed between inflations. After each left atrial balloon inflation, neutrophil uptake remained unchanged from base line, despite decreased mean cardiac output to 0.67 +/- 0.24 (+/- SD) 1/min and increased pulmonary blood volume. The absence of pulmonary neutrophil uptake was confirmed by arterial-venous measurements. Increased pulmonary blood volume had little effect on lung neutrophil uptake, suggesting that most of the pulmonary neutrophils are marginated. We conclude that the lungs have a large marginated neutrophil pool compared with the circulating pool and that reduced cardiac output and elevated left atrial pressure have no effect on pulmonary neutrophil kinetics in the sheep.     

Effects of reduction on the denaturation kinetics of human hair

Although human hair as an alpha-keratinous fiber exhibits a complex morphology, it can be considered as a nano-structured filament/matrix composite for the context of thermal analysis. Using differential scanning calorimetry (DSC) in water, the denaturation performance of the alpha-helical protein fraction and the effects of reductive treatments were studied. The results are viewed in the context of a previous study for oxidative treatments. It was found that the course of the denaturation process remains generally unperturbed by the treatment, following an irreversible, one-step, first-order process. Arrhenius activation energies and pre-exponential factors were determined from the DSC-curves by applying the principles of the Friedman-method. Comparing activation energy values between reductive and oxidative processes shows the differences of the effects on the components of the composite. In contrast, the values of the rate constant at the denaturation temperature, though showing differences in their trends with cumulative treatments, are very similar. This further emphasizes the theory that the viscosity of the matrix affects strict kinetic control over the denaturation of the alpha-helical segments. Once the viscosity of the matrix has decreased enough for the denaturation process to occur, this follows a path that is largely independent of the temperature range and of the chemical pre-history.     

Expression profile analysis of microRNAs during hair follicle development in the sheep foetus

MicroRNAs (miRNAs) regulate the development and growth cycle of hair follicles (HFs). The molecular mechanism by which miRNAs determine the development of HFs in the sheep foetus remains elusive. In this study, the expression profiles of miRNAs at 11 development periods (45, 55, 65, 75, 85, 95, 105, 115, 125, 135 and 145 d) in sheep foetus skin were analysed by high-throughput sequencing and bioinformatics analysis. A total of 72 conserved miRNAs, 44 novel miRNAs and 32 known miRNAs were significantly differentially expressed. qRT-PCR results for 18 miRNAs were consistent with the sequencing data. 85 d of foetal development was the starting point for secondary hair follicle (SF) development according to tissue morphology and cluster analysis. In SF development, the prolactin signalling pathway and platelet activation played important roles, and 10 miRNAs were potential candidate miRNAs in SF initiation.