Heat processing has been used to improve protein utilization and availability of animal nutrition. However, to date, few studies exist on heat-induced protein molecular structure changes on a molecular basis. The aims of this study were to use molecular spectroscopy as a novel approach to determine heat-induced protein molecular structure changes affected by moist and dry heating and quantify protein molecular structures and nutritive value in the rumen and intestine in dairy cattle. In this study, soybean was used as a model for feed protein and was autoclaved at 120°C for 1h (moist heating) and dry heated at 120°C for 1h. The parameters assessed in this study included protein structure α-helix and β-sheet and their ratio, protein subfractions associated with protein degradation behaviors, intestinal protein availability, and energy values. The results show that heat treatments changed the protein molecular structure. Both dry and moist heating increased the amide I-to-amide II ratio. However, for the protein α-helix-to-β-sheet ratio, moist heating decreased but dry heating increased the ratio. Compared with dry heating, moist heating dramatically changed the chemical and nutrient profiles of soybean seed. It greatly decreased soluble crude protein, nonprotein nitrogen, and increased neutral detergent insoluble protein. Both dry and moist heating treatments did not alter digestible nutrients and energy values. Heating tended to decrease the nonprotein nitrogen fraction (soluble and rapidly degradable protein fraction) and true protein 1 fraction (fast-degradable protein fraction). Conversely, the true protein 3 fraction (slowly degradable fraction) significantly increased. The in situ rumen study showed that moist heating decreased protein rumen degradability and increased intestinal digestibility of rumen-undegradable protein. Compared with the raw soybeans, dry heating did not affect rumen degradability and intestinal digestibility. In conclusion, compared with dry heating, moist heating dramatically affected the nutrient profile, protein subfractions, rumen degradability, intestinal digestibility, and protein molecular structure (amide I-to-II ratio; α-helix-to-β-sheet ratio). The sensitivity of soybean seed to moist heating was much higher than that to dry heating in terms of the structure and nutrient profile changes.

1. Experiments were conducted to establish the requirements and optimal dietary ratio of lysine to threonine for fast growing male chickens (genotype Ross 308) depending on age, daily protein deposition and of dietary amino acid efficiency. 2. A total of 216 growing chickens were utilised in nitrogen-balance studies in three age periods (10 to 25 d; 30 to 45 d; 50 to 65 d) using graded levels of protein supply (60 to 360 g/kg crude protein) in lysine or threonine limiting diets. 3. Supplementation of crystalline amino acids (L-Lys, L-Thr, DL-Met and L-Arg) provided the following amino acid ratios: lysine limiting diets (Lys:Met + Cys:Thr:Arg = 1 : 1.01 : 0.91 : 1.14), threonine limiting diets (Lys : Met + Cys : Thr : Arg = 1 : 0.85 : 0.54 : 1.16). 4. The principles of the diet dilution technique using an exponential function were applied for the modelling of lysine and threonine requirements. For equal daily protein deposition, optimal lysine to threonine ratios 1 : 0.69 (10 to 25 d), 1 : 0.70 (30 to 45 d) and 1 : 0.74 (50 to 65 d) were established. 5. For the commercial growth period of fast growing chickens, the derived optimal lysine to threonine ratio was constant (1 : 0.69). The applied modelling procedure gave conclusions for quantitative requirements and optimal dietary lysine:threonine ratios in line with actual recommendations.

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N-balance studies were carried out to assess the lysine requirement of fast growing chickens (Cobb /**) at di#erent sex and age depending on crude protein deposition and e$ciency of dietary lysine utilisation. The experiments were conducted within three age periods (I : +*ῌ,/ d ; II :-*ῌ./ d ; III : /*ῌ0/ d) and 1, chickens (-0 males,-0 females) per age period. Experimental diets with six levels of graded CP content were based on high protein (HP)-soybean meal, wheat gluten and crystalline amino acids (L-Thr, DL-Met, L-Arg) in order to create lysine (..-* g Lys/+** gCP) as the first limiting dietary amino acid (constant ratio Lys : MetῌCys : Thr : Arg῍+ : +.*+ : *.3+ : +.+.). For application of a nonlinear N-utilization model, nitrogen maintenance requirement (NMR) and theoretical maximum for daily nitrogen retention (NRmaxT) were established as model parameters for further assessment of the lysine requirement depending on age, sex and daily CP-deposition. As an example, the calculated lysine requirement concentration for 0*ῌ of the theoretical potential for daily CP-deposition (+*ῌ,/ d : +.+*ῌ lysine, 0* g daily feed intake ;-*ῌ./ d : +.*-ῌ lysine, +.* g daily feed intake ; /*ῌ0/ d : *.30ῌ lysine, +1* g daily feed intake) was in close agreement with published data. However, the predicted feed intake is one of the most important factors of influence when amino acid requirement concentrations are established. The level of daily CP-deposition and the dietary amino acid e$ciency as important factors influencing the amino acid requirement data need more attention in future requirement studies.

Experiments were conducted to estimate daily N maintenance requirements (NMR) and the genetic potential for daily N deposition (ND(max)T) in fast-growing chickens depending on age and sex. In N-balance studies, 144 male and 144 female chickens (Cobb 500) were utilized in 4 consecutive age periods (I: 10 to 25 d; II: 30 to 45 d; III: 50 to 65 d; and IV: 70 to 85 d). The experimental diets contained high-protein soybean meal and crystalline amino acids as protein sources and 6 graded levels of protein supply (N1 = 6.6%; N2 = 13.0%; N3 = 19.6%; N4 = 25.1%; N5 = 31.8%; and N6 = 37.6% CP in DM). The connection between N intake and total N excretion was fitted for NMR determination by an exponential function. The average NMR value (252 mg of N/BW(kg)0.67 per d) was applied for further calculation of ND(max)T as the threshold value of the function between N intake and daily N balance. For estimating the threshold value, the principle of the Levenberg-Marquardt algorithm within the SPSS program (Version 11.5) was applied. As a theoretical maximum for ND(max)T, 3,592, 2,723, 1,702, and 1,386 mg of N/BW(kg)0.67 per d for male and 3,452, 2,604, 1,501, and 1,286 mg of N/BW(kg)0.67 per d for female fast-growing chickens (corresponding to age periods I to IV) were obtained. The determined model parameters were the precondition for modeling of the amino acid requirement based on an exponential N-utilization model and depended on performance and dietary amino acid efficiency. This procedure will be further developed and applied in the subsequent paper.

Nitrogen-balance experiments were conducted with a total of 288 male chickens to assess Thr requirement data on 2 commercial slow-growing genotypes (I 657 and Red JA from Hubbard ISA) by use of a modeling procedure described previously. Six graded levels of dietary protein supply from high-protein soybeanmeal were used within 4 age periods (period I: 10 to 25 d; period II: 30 to 45 d; period III: 5 to 65 d; and period IV: 70 to 85 d). The provided dietary amino acid ratio (Lys:Met+Cys:Thr=1:0.85:0.54), with 3.87% Thr in the feed protein, identified Thr as the first limiting dietary amino acid. The nitrogen maintenance requirement (NMR) was established by exponential approximation of N excretion depending on N intake (on average, NMR=173 mg of N/BWkg0.67 per d). The theoretical maximum for daily N deposition was estimated by the Levenberg-Marquardt algorithm (SPSS program, version 11.5) and by exponential fitting of N balance data depending on N intake. The observed dietary Thr efficiency was used to model Thr requirements for a given protein deposition depending on age. The optimal dietary Thr concentration (percentage of feed) was established by different predictions for daily feed intake. Daily CP deposition of approximately 60% of the potential required 0.83 and 0.87% (10 to 25 d), 0.73 and 0.75% (30 to 45 d), 0.66 and 0.69% (50 to 65 d), and 0.51 and 0.53% (70 to 85 d) of Thr in feed for genotype I 657 and genotype Red JA, respectively (average daily feed intakes of 30, 75, 100, and 100 g in age periods I to IV). Results of model calculations need verification in comparative growth studies with assessment of nutrient deposition and varying dietary Thr efficiencies.

Objective To explore the utility of intraoperative cavernosal nerve stimulation in facilitating atraumatic nerve dissection during radical prostatectomy, and thus help predict postoperative erectile function. Patients and methods Fourteen patients (aged 51–72 years) underwent nerve‐sparing radical retropubic prostatectomy (NSRRP); 10 were potent before surgery (group 1), and four had erectile dysfunction (group 2). A multi‐acquisition system (MacLab/8e) with a Macintosh computer was used for real‐time display and recording of intracavernosal pressure (ICP) during surgery. Nerves were stimulated with a bipolar probe (monophasic rectangular pulses, 10 mA, 20 Hz, 0.22 s) before and after removal of the gland. The follow‐up consisted of interviews with patients and their partners’ 12–18 months after treatment. Results The mean (sem) basal ICP of 8.0 (2.0) cmH2O remained unchanged during nerve dissection. The mean increase in ICP during electrical stimulation was >50 cmH2O in seven potent patients (group 1) and was sustained as long as the nerve was stimulated. Postoperatively, these seven patients reported erections sufficient for sexual intercourse. However, the three remaining patients in group 1 had pressure rises of <30 cmH2O, of whom two reported partial erections and one reported total impotence postoperatively. The recovery time for erectile function was 6–12 months after surgery. Two patients from group 2 had transient increases in ICP to <40 cmH2O; one had an increase to 20 cmH2O and one had no response at all. All four patients remained totally impotent postoperatively. There were no complications. Conclusions Intraoperative electrical stimulation of the cavernosal nerves with ICP monitoring before and after NSRRP is a safe and reliable method for documenting nerve continuity and its functional status. Patients who have normal preoperative erectile function and show an adequate rise in ICP upon electrical nerve stimulation during NSRRP will almost certainly be potent after surgery. This tool may be used to facilitate atraumatic nerve dissection during NSRRP.

In addition to dose-response studies, modeling of N utilization, depending on intake of the first limiting amino acid in the diet, is one of the tools for assessing amino acid requirements in growing animals. Based on a verified nonlinear N-utilization model and following the principles of the diet dilution technique, N-balance experiments were conducted to estimate the Thr requirement of fast-growing chickens (genotype Cobb), depending on age, sex, CP deposition. and efficiency of dietary Thr utilization. Different predictions were made for the feed intake to conclude the optimal Thr concentration in the feed. The results are based on N-balance experiments with a total of 144 male and 144 female growing chickens within 4 age periods (I: 10 to 25 d; II: 30 to 45 d; III: 50 to 65 d; IV: 70 to 85 d), using diets with graded protein supply (6.6, 13, 19.6, 25.1, 31.8, and 37.6% CP in DM) from high-protein soybean meal with a constant amino acid ratio and Thr as the first limiting amino acid (3.87 g of Thr/100 g of CP; dietary Lys:Thr = 1:0.54). The observed optimal Thr concentration (% of feed) was influenced by age, sex, level of CP deposition, dietary efficiency of Thr utilization, and predicted feed intake. For male chickens, assuming an average CP deposition (60% of the potential) and average efficiency of Thr utilization, 0.78% (10 to 25 d), 0.73% (30 to 45 d), 0.65% (50 to 65 d), and 0.55% (70 to 85 d) total dietary Thr were observed as optimal total Thr concentration in the diet (corresponding to 60, 135, 160, and 180 g of daily feed intake, respectively). Data are discussed in context with the main factors of influence like age, sex, level of daily CP deposition, efficiency of dietary Thr utilization, and predicted feed intake.