Stabilizing isopeptide bonds revealed in gram-positive bacterial pilus structure

Many bacterial pathogens have long, slender pili through which they adhere to host cells. The crystal structure of the major pilin subunit from the Gram-positive human pathogen Streptococcus pyogenes at 2.2 angstroms resolution reveals an extended structure comprising two all-beta domains. The molecules associate in columns through the crystal, with each carboxyl terminus adjacent to a conserved lysine of the next molecule. This lysine forms the isopeptide bonds that link the subunits in native pili, validating the relevance of the crystal assembly. Each subunit contains two lysine-asparagine isopeptide bonds generated by an intramolecular reaction, and we find evidence for similar isopeptide bonds in other cell surface proteins of Gram-positive bacteria. The present structure explains the strength and stability of such Gram-positive pili and could facilitate vaccine development.     

A high-resolution record of holocene climate change in speleothem calcite from cold water cave, northeast iowa

High-precision uranium-thorium mass spectrometric chronology and (18)O-(13)C isotopic analysis of speleothem calcite from Cold Water Cave in northeast Iowa have been used to chart mid-Holocene climate change. Significant shifts in dagger(18)O and dagger(13)C isotopic values coincide with well-documented Holocene vegetation changes. Temperature estimates based on (18)O/(16)O ratios suggest that the climate warmed rapidly by about 3 degrees C at 5900 years before present and then cooled by 4 degrees C at 3600 years before present. Initiation of a gradual increase in dagger(13)C at 5900 years before present suggests that turnover of the forest soil biomass was slow and that equilibrium with prairie vegetation was not attained by 3600 years before present.     

Reducing chilling injury of potatoes by intermittent warming

Potato tubers held continuously 19 weeks at 32 F developed chillinginjury symptoms early during storage and the symptoms worsened with time in storage. Early symptoms included browning and surface mold on skinned areas. By 11 weeks mahogany browning, blackheart, and hollow heart, bluish skin discoloration and sinking of intact skin were visible in daylight. And under ultraviolet light, yellow fluorescence was visible in halos, washes, or spots around or near internal tuber tissues discolored by other chilling-injury symptoms. Some damage at 32 F became visible after subsequent holding of tubers for 1 week at 60 F. Tubers accumulated high amounts of reducing and total sugar at 32 F and respiration, measured at 60 F as CO₂ evolved, was progressively stimulated by time at 32 F. Sugar build-up, respiration-rate increase, and chilling injury seemed to show no cause-and-effect relationship but appeared as separate phenomena each caused by the stress of holding tubers at too low a temperature. Moving potatoes to 60 F for 1 week after every 3rd week to interrupt storage at 32 practically eliminated all forms of chilling injury except skin browning and surface mold which were greatly reduced. Sugar content and respiration rates in tubers intermittently held at low and high temperatures were higher after periods in low temperature and lower after periods at 60 F, and in general were maintained at levels much lower than in potatoes held continuously at 32, 36, or 40 F. In all treatments sugar content was higher than desirable for chipping type potatoes. Holding potatoes at 60 F for 1 to 4 weeks before placing them at 32 F reduced skin browning and surface mold but had no effect on other forms of chilling injury, sugar accumulation, or respiration rates in tubers as measured after 15 to 19 weeks continuously at 32 F. Practically no chilling-injury symptom except for very little mold growth and browning of skinned areas developed in tubers held in 36 or 40 F.     

Historical and Modern Disturbance Regimes,Stand Structures,and Landscape Dynamics in Piñon–Juniper Vegetation of the Western United States

Piñon–juniper is a major vegetation type in western North America. Effective management of these ecosystems has been hindered by inadequate understanding of 1) the variability in ecosystem structure and ecological processes that exists among the diverse combinations of piñons, junipers, and associated shrubs, herbs, and soil organisms; 2) the prehistoric and historic disturbance regimes; and 3) the mechanisms driving changes in vegetation structure and composition during the past 150 yr. This article summarizes what we know (and don't know) about three fundamentally different kinds of piñon–juniper vegetation. Persistent woodlands are found where local soils, climate, and disturbance regimes are favorable for piñon, juniper, or a mix of both; fires have always been infrequent in these woodlands. Piñon–juniper savannas are found where local soils and climate are suitable for both trees and grasses; it is logical that low-severity fires may have maintained low tree densities before disruption of fire regimes following Euro-American settlement, but information is insufficient to support any confident statements about historical disturbance regimes in these savannas. Wooded shrublands are found where local soils and climate support a shrub community, but trees can increase during moist climatic conditions and periods without disturbance and decrease during droughts and following disturbance. Dramatic increases in tree density have occurred in portions of all three types of piñon–juniper vegetation, although equally dramatic mortality events have also occurred in some areas. The potential mechanisms driving increases in tree density—such as recovery from past disturbance, natural range expansion, livestock grazing, fire exclusion, climatic variability, and CO₂ fertilization—generally have not received enough empirical or experimental investigation to predict which is most important in any given location. The intent of this synthesis is 1) to provide a source of information for managers and policy makers; and 2) to stimulate researchers to address the most important unanswered questions.     

Effect of hCG Treatment on the Oestrous and Ovulation Responses to FSH in Prepubertal Gilts

To ensure sufficient numbers of pregnant females, particularly at hotter times of the year, hormonal induction of gilt oestrus may be necessary. However, the gilt oestrus and ovulation responses to gonadotrophin treatment have often proven unpredictable. The objective of this study was to examine possible reasons for this unpredictability. Prepubertal gilts (approximately 150 days of age, n = 63) were assigned to one of three treatments: injection of 300 IU hCG (n = 15); pre-treatment with 100 mg FSH in polyvinylpyrrolidinone administered as 2 × 50 mg injections 24 h apart, followed by 600 IU eCG at 24 h after the second FSH injection (n = 23); or FSH pre-treatment as above followed by 300 IU hCG at 24 h after the second FSH injection (n = 25). To facilitate oestrus detection, gilts were exposed to a mature boar for 15 min daily for 7 days. Blood samples were obtained on the day of eCG or hCG injection and again 10 days later and gilt ovulation responses determined based on elevated progesterone concentrations. The oestrus responses by 7 days were 6.7%, 17.5% and 64.0% for gilts treated with hCG, FSH + eCG and FSH + hCG, respectively (p < 0.001). The oestrous gilt receiving hCG alone and one oestrous FSH + hCG gilt did not ovulate, all other oestrous gilts ovulated. A further two anoestrous FSH + eCG-treated gilts ovulated. These data suggest that FSH pre-treatment facilitated the development of ovarian follicles to the point where they became responsive to hCG, but had little effect on the response to eCG.     

The effect of calcium and phosphorus supplementation, inactivity, and subsequent aerobic training on the mineral balance in young, mature, and aged horses

A randomized complete block design experiment was conducted to determine the effect of Ca and P supplementation, inactivity, and subsequent aerobic exercise on Ca and P serum concentrations, and balance and digestibilities in young, mature, and aged horses. Twelve stock-type geldings were blocked into three age groups: 2 and 3, 7 to 11, and 15 to 21 years of age. Two groups of six horses were randomly assigned to either control (C) or mineral supplemented (T) diet. Horses consuming C received about 133% of National Research Council (NRC) requirements for Ca and P, while horses consuming T received about 275% of NRC requirements for Ca and P. The trial consisted of eight 21-d periods. During periods I, II, and III, all horses were inactive, and during periods IV, V, VI, VII, and VIII, all horses performed an ascending aerobic exercise regimen. Serum samples were taken at the beginning of period I and at 21-d intervals thereafter for determination of serum Ca and P. Total fecal and urine collections were taken for 72 h at the beginning of period I and at 21-d intervals thereafter for determination of Ca and P balance. Overall mean apparent Ca balance was affected by treatment (P < .009); time (P < .001); and the interaction of time and treatment (P < .001). Overall mean apparent Ca balance was higher (P < .009) for T as compared to C (23.63 vs. 3.77 mg/kg BW). Mean apparent Ca digestibility was affected by time (P < .001) and the interaction of time and age (P < .001). Mean apparent P balance was affected by time (P < .001) and the interaction of time and treatment (P < .032). Increases in Ca balance may indicate more available mineral for bone metabolism, particularly when remodeling is triggered with the onset of exercise following a sedentary period, suggesting a possible benefit to feeding additional Ca above current NRC recommendations.     

Use of the novel atypical opioid tapentadol in goats (Capra hircus): pharmacokinetics after intravenous,and intramuscular administration

The objective of the present study was to assess the pharmacokinetics of the novel atypical drug tapentadol (TAP) after intravenous (I.V.) and intramuscular (I.M.) injections in clinically healthy goats. A 2 × 2 cross‐over design study was carried out. Six local adult Nubian nonlactating, nonpregnant female goats, were given 5 mg/kg body weight of TAP by I.V. and I.M. routes. The concentrations of TAP in plasma were evaluated using a validated HPLC method. Transient adverse effects were noticed in some animals, especially after I.V. administration (tremors and ataxia). Three days after drug administration, severe hair loss was also recorded. The plasma concentrations after the two routes of administration were best described by a bi‐compartmental model. After I.M. injection, TAP showed a very fast absorption (T_max = 0.17 h) and a short half‐life (1.29 h). The I.M. bioavailability was quite high, despite being variable (87.8 ± 35.6%). This is the first pharmacokinetic study of TAP in goats but due to its unknown safety profile and efficacy, it is premature to recommend the use of this drug in clinical ovine practice.