An integrative model of amino acid metabolism in the liver of the lactating dairy cow

The objective of this work was to construct a dynamic model of hepatic amino acid metabolism in the lactating dairy cow that could be parameterized using net flow data from in vivo experiments. The model considers 22 amino acids, ammonia, urea, and 13 energetic metabolites, and was parameterized using a steady-state balance model and two in vivo, net flow experiments conducted with mid-lactation dairy cows. Extracellular flows were derived directly from the observed data. An optimization routine was used to derive nine intracellular flows. The resulting dynamic model was found to be stable across a range of inputs suggesting that it can be perturbed and applied to other physiological states. Although nitrogen was generally in balance, leucine was in slight deficit compared to predicted needs for export protein synthesis, suggesting that an alternative source of leucine (e.g. peptides) was utilized. Simulations of varying glucagon concentrations indicated that an additional 5 mol/d of glucose could be synthesized at the reference substrate concentrations and blood flows. The increased glucose production was supported by increased removal from blood of lactate, glutamate, aspartate, alanine, asparagine, and glutamine. As glucose output increased, ketone body and acetate release increased while CO(2) release declined. The pattern of amino acids appearing in hepatic vein blood was affected by changes in amino acid concentration in portal vein blood, portal blood flow rate and glucagon concentration, with methionine and phenylalanine being the most affected of essential amino acids. Experimental evidence is insufficient to determine whether essential amino acids are affected by varying gluconeogenic demands.     

Reducing dietary protein in dairy cow diets: implications for nitrogen utilization,milk production,welfare and fertility

In light of increasing global protein prices and with the need to reduce environmental impact of contemporary systems of milk production, the current review seeks to assess the feasibility of reducing levels of dietary CP in dairy cow diets. At CP levels between 140 and 220 g/kg DM there is a strong positive relationship between CP concentration and dry matter intake (DMI). However, such effects are modest and reductions in DMI when dietary CP is below 180 g/kg DM can be at least partially offset by improving the digestibility and amino acid profile of the undegradable protein (UDP) component of the diet or by increasing rumen fermentable energy. Level and balance of intestinally absorbable amino acids, in particular methionine and lysine, may become limiting at lower CP concentrations. In general the amino acid composition of microbial protein is superior to that of UDP, so that dietary strategies that aim to promote microbial protein synthesis in the rumen may go some way to correcting for amino acid imbalances in low CP diets. For example, reducing the level of NDF, while increasing the proportion of starch, can lead to improvements in nitrogen (N) utilisation as great as that achieved by reducing dietary CP to below 150 g/kg. A systematic review and meta-analysis of responses to rumen protected forms of methionine and lysine was conducted for early/mid lactation cows fed diets containing ⩽150 g CP/kg DM. This analysis revealed a small but significant (P=0.002) increase in milk protein yield when cows were supplemented with these rumen protected amino acids. Variation in milk and milk protein yield responses between studies was not random but due to differences in diet composition between studies. Cows fed low CP diets can respond to supplemental methionine and lysine so long as DMI is not limiting, metabolisable protein (MP) is not grossly deficient and other amino acids such as histidine and leucine do not become rate limiting. Whereas excess dietary protein can impair reproduction and can contribute to lameness, there is no evidence to indicate that reducing dietary CP levels to around 140 to 150 g CP/kg DM will have any detrimental effect on either cow fertility or health. Contemporary models that estimate MP requirements of dairy cows may require refinement and further validation in order to predict responses with low CP diets.     

Dietary influence on protein level in milk and milk yield in dairy cows

Diet can influence the yield of milk protein more than it can influence milk protein content. Providing sufficient dietary crude protein in forms that will maximize amounts of amino acids required for milk protein synthesis to the gastrointestinal tract, in forms that can be digested and absorbed, will provide for optimal milk protein yield and content. Maximizing ruminal microbial protein synthesis is an important part of this strategy. The rest of the strategy involves providing sufficient amounts of the remaining required amino acids as ruminally protected proteins, or amino acids in forms that can be digested in the gastrointestinal tract. A diet deficient in protein will reduce milk protein content 1 to 2 g kg⁻¹ and may substantially reduce yields of milk and milk protein. A diet containing high amounts of readily fermentable carbohydrates may increase milk protein content 1 to 2 g kg⁻¹, and may increase yields of milk and protein, but may also result in digestive and metabolic upsets. Diets containing supplemental fat will increase yield of milk protein, but not as extensively as the increase in yield of milk, because milk protein content is usually reduced 1 to 2 g kg⁻¹. The increased efficiency of milk fat and lactose synthesis is likely to be the reason for this depression in milk protein content. A means of overcoming this problem is a continuing research challenge.     

Using pseudo amino acid composition to predict protein subcellular location: approached with amino acid composition distribution

In the Post Genome Age, there is an urgent need to develop the reliable and effective computational methods to predict the subcellular localization for the explosion of newly found proteins. Here, a novel method of pseudo amino acid (PseAA) composition, the so-called “amino acid composition distribution” (AACD), is introduced. First, a protein sequence is divided equally into multiple segments. Then, amino acid composition of each segment is calculated in series. After that, each protein sequence can be represented by a feature vector. Finally, the feature vectors of all sequences thus obtained are further input into the multi-class support vector machines to predict the subcellular localization. The results show that AACD is quite effective in representing protein sequences for the purpose of predicting protein subcellular localization.     

Use of physiological parameters to predict milk yield and feed intake in heat-stressed dairy cows

Holstein cows were tested to determine which measures of thermal strain are the better predictors of production over 24–96 h at constant air temperatures of 19 and 29 °C. Both rectal temperature and respiration rate increased within 24 h, followed by milk yield and feed intake reductions after 48 h of heat stress. There were significant correlations between milk yield, feed intake, and rectal temperature. Several physiological strain indices were created to determine if combinations of measures are better than single measures in predicting production under these acute conditions. Mean daily rectal temperature was superior to maximum and minimum daily values of rectal temperature, as well as other indicators of thermal status in predicting dairy cow production. Likewise, mean daily rectal temperature was equal if not better than the physiological strain indices used in this study in predicting production. These data suggest that rectal temperature is superior to both single and combined indicators of thermal status in predicting dairy cow production, and should be considered for future development of physiological strain indices over a longer time period in both laboratory and field environments.     

Restricting dairy cow access time to pasture in early lactation: the effects on milk production, grazing behaviour and dry matter intake

One of the main aims of pasture-based systems of dairy production is to increase the proportion of grazed grass in the diet. This is most easily achieved by increasing the number of grazing days. However, periods of inclement weather conditions can reduce the number of days at pasture. The two objectives of this experiment were: (i) to investigate the effect of restricting pasture access time on animal production, grazing behaviour and dry matter intake (DMI) of spring calving dairy cows in early lactation; and (ii) to establish whether silage supplementation is required when cows return indoors after short grazing periods. In all, 52 Holstein-Friesian spring calving dairy cows were assigned to a four-treatment study from 25 February to 26 March 2008. The four treatments were: full-time access to pasture (22H; control); 4.5-h- pasture access after both milkings (2 × 4.5H); 3-h pasture access after both milkings (2 × 3H); 3-h pasture access after both milkings with silage supplementation by night (2 × 3SH). All treatments were offered 14.4 kg DM/cow per day herbage from swards, with a mean pre-grazing yield of 1739 kg DM/ha above 4 cm, - and were supplemented with 3 kg DM/cow per day of concentrate. The 2 × 3SH treatment was offered an additional 4 kg DM/cow of grass silage by night. Restricting pasture access time (2 × 3H, 2 × 3SH and 2 × 4.5H) had no effect on milk (28.3 kg/cow per day) and solids-corrected milk (27.2 kg/cow per day) yield when compared with the treatment grazing full time. Supplementing animals with grass silage did not increase milk production when compared with all other treatments. Milk protein concentration tended to be lower (P = 0.08; 32.2 g/kg) for the 2 × 3SH animals when compared with the 22H animals (33.7 g/kg). The grass DMI of the 2 × 3SH treatment was significantly lower (-2.3 kg DM/cow per day) than all other treatments (11.9 kg DM/cow per day), yet the total DMI of these animals was highest (16.6 kg DM/cow per day). The 22H cows grazed for 481 min/cow per day, which is significantly longer than all other treatments. The 2 × 3H animals grazed for 98% of the time, whereas the 2 × 3SH grazed for 79% of their time at pasture. Restricting pasture access time did not affect end body weight or body condition score. The results of this study indicate that restricting pasture access time of dairy cows in early lactation does not affect milk production performance. Furthermore, supplementing cows with grass silage does not increase milk production but reduces grazing efficiency.     

Restricting dairy cow access time to pasture in autumn: The effects on milk production,grazing behaviour and DM intake of late lactation dairy cows

Extending the grazing season in pasture based systems of dairy production can increase farm profitability; poor weather and soil conditions can reduce the number of grazing days. The study objectives were to (i) examine the effect of restricted access to pasture in the autumn on the milk production, grazing behaviour and DM intake (DMI) of late lactation spring-calving dairy cows and (ii) establish the effect of alternating restricted and continuous access to pasture on dairy cow production, DMI and grazing behaviour. Cows were randomly assigned to one of four grazing treatments: (i) 22 h (full-time) access to pasture (22H; control); (ii) Two 5-h periods of access to pasture (2×5H); (iii) Two 3-h periods of access to pasture (2×3H); and (iv) alternating between full-time and 3-h access to pasture with no more than three continuous days on any one regime, e.g. Monday – full-time access, Tuesday − 2x3H access, Wednesday − 2x3H access; Thursday – full-time access, etc. (2×3HV). Restricted access to pasture was offered after a.m. and p.m. milking. Swards of similar quality and pregrazing herbage mass were offered. Treatment had no effect on milk yield (13.2 kg/day), milk fat (48.2 g/kg), protein (39.0 g/kg) or lactose content (42.6 g/kg) and milk solid yield (1.15 kg/day). Similarly, there was no effect of treatment on final BW (483 kg) or final BCS (2.66). There was no significant difference in DMI (15.1 kg DM/cow/day) between treatments. There was an effect on daily grazing time, 22H cows (565 min/cow/day) grazed for longest time, however, when the 2x3HV treatment had full-time access to pasture, they had a similar grazing time (543 min/cow/day) to the 22H cows and were similar to the 2x3H treatment on days with restricted access to pasture (357 min/cow/day). The 22H and 2x5H animals had similar grass DMI/min (29.2 g/min), the 2x3HV were higher (33.9 g/min) but were similar to the comparable treatment when offered 2x3H access time (41.6 g/min) and when offered 22H access time (27.7 g/min). The results from this study show how when offered a grass only diet of autumn pasture grazing behaviour can be modified by restricting pasture access time without reducing dairy cow production in late lactation at low production levels. There was also no effect of alternating access time between 22H and 2x3H on milk production and DMI in the 2x3HV treatment. Restricted access time to pasture in autumn may be a strategy which farmers can use to extend the grazing season.     

High- and low-purity glycerine supplementation to dairy cows in early lactation: effects on silage intake,milk production and metabolism

This study evaluated the effects of supplemental low- and high-purity glycerine on silage intake, milk yield and composition, plasma metabolites and body condition score (BCS) in dairy cows. A total of 42 cows of the Swedish Red Breed, housed in individual tie stalls, were fed 0.25 kg of low- or high-purity glycerine on top of concentrate, twice daily, during the first 4 weeks of lactation. One-third of the cows acted as controls, receiving no glycerine. Silage was fed for ad libitum intake and concentrate was fed at restricted level of intake, about 6 kg/day for primiparous cows and 7 kg/day for multiparous cows. Feed refusals were weighed daily. Cows were milked twice daily, milk yield was recorded on four occasions per week and milk samples were collected simultaneously. Blood samples were drawn from the coccygeal vessel once a week. Low- and high-purity glycerine had no effect on silage or total dry matter intake (P = 0.38 and P = 0.75, respectively) or on BCS (P = 0.45). Cows fed high-purity glycerine tended to have higher milk yield than control cows (P = 0.06). Milk composition tended to differ among treatments. No main effects of treatment on concentration of glycerine (P = 0.44), glucose (P = 0.78), insulin (P = 0.33), non-esterified fatty acids (P = 0.33) and β-hydroxybutyrate (P = 0.15) in plasma. These data indicate that high-purity glycerine has the potential to increase milk yield, as well as enhance the milk protein concentration and milk fat + protein yield.     

Effect of butaphosphan and cyanocobalamin on postpartum metabolism and milk production in dairy cows

The aim of this study was to determine the effect of butaphosphan and cyanocobalamin (BTPC) supplementation on plasma metabolites and milk production in postpartum dairy cows. A total of fifty-two Holstein cows were randomly assigned to receive either: (1) 10 ml of saline (NaCl 0.9%, control group); (2) 1000 mg of butaphosphan and 0.5 mg of cyanocobalamin (BTPC1 group); and (3) 2000 mg of butaphosphan and 1.0 mg of cyanocobalamin (BTPC2 group). All cows received injections every 5 days from calving to 20 days in milk (DIM). Blood samples were collected every 15 days from calving until 75 DIM to determine serum concentration of glucose, non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHB), cholesterol, urea, calcium (Ca), phosphorus (P), magnesium (Mg), aminotransferase aspartate (AST) and γ-glutamyltransferase (GGT). The body condition score (BCS) and milk production were evaluated from calving until 90 DIM. Increasing doses of BTPC caused a linear reduction in plasma concentrations of NEFA and cholesterol. Supplementation of BTPC also reduced concentrations of BHB but it did not differ between the two treatment doses. Milk yield and milk protein had a linear increase with increasing doses of BTPC. A quadratic effect was detected for milk fat and total milk solids according to treatment dose, and BTPC1 had the lowest mean values. Concentrations of glucose, urea, P, Mg, AST, GGT, milk lactose and BCS were not affected by treatment. These results indicate that injections of BTPC during the early postpartum period can reduce NEFA and BHB concentrations and increase milk production in Holstein cows.     

An individual reproduction model sensitive to milk yield and body condition in Holstein dairy cows

To simulate the consequences of management in dairy herds, the use of individual-based herd models is very useful and has become common. Reproduction is a key driver of milk production and herd dynamics, whose influence has been magnified by the decrease in reproductive performance over the last decades. Moreover, feeding management influences milk yield (MY) and body reserves, which in turn influence reproductive performance. Therefore, our objective was to build an up-to-date animal reproduction model sensitive to both MY and body condition score (BCS). A dynamic and stochastic individual reproduction model was built mainly from data of a single recent long-term experiment. This model covers the whole reproductive process and is composed of a succession of discrete stochastic events, mainly calving, ovulations, conception and embryonic loss. Each reproductive step is sensitive to MY or BCS levels or changes. The model takes into account recent evolutions of reproductive performance, particularly concerning calving-to-first ovulation interval, cyclicity (normal cycle length, prevalence of prolonged luteal phase), oestrus expression and pregnancy (conception, early and late embryonic loss). A sensitivity analysis of the model to MY and BCS at calving was performed. The simulated performance was compared with observed data from the database used to build the model and from the bibliography to validate the model. Despite comprising a whole series of reproductive steps, the model made it possible to simulate realistic global reproduction outputs. It was able to well simulate the overall reproductive performance observed in farms in terms of both success rate (recalving rate) and reproduction delays (calving interval). This model has the purpose to be integrated in herd simulation models to usefully test the impact of management strategies on herd reproductive performance, and thus on calving patterns and culling rates.     

Structured model to predict intracellular amino acid shortages during recombinant protein overexpression in E. coli.

Recombinant protein overexpression and the classical stringent response have been shown to induce the same proteases. Since the stringent response was the result of an intracellular amino acid shortage, it was hypothesized that the overexpression of the recombinant protein also caused an intracellular amino acid shortage. A structured non-segregated kinetic mathematical model of recombinant Escherichia coli was developed to predict intracellular amino acid shortages during recombinant protein overexpression, and thus the induction of the stringent response. Two model recombinant proteins were examined, chloramphenicol acetyl-transferase (CAT) and an 'average protein'. The model predicted an aromatic amino acid shortage during CAT overexpression, as predicted based on the CAT's amino acid content. The model also predicted a shortage of the intracellular alanine family amino acid pool during CAT overexpression. This was unexpected due to the relatively low content of alanine family amino acids in CAT compared to the average E. coli protein. The model predicted alanine, glutarate, and aspartate family amino acid shortages during recombinant 'average protein' overexpression. Additionally, the model predicted a decrease in the ribosome pool at induction for both recombinant proteins, which agrees with published experimental results. Thus, the structured kinetic model was able to predict amino acid shortages, that could potentially cause a stringent response and elevated protease activity.     

Erratum to: Rumen-protected conjugated linoleic acid supplementation to dairy cows in late pregnancy and early lactation: effects on milk composition,milk yield,blood metabolites and gene expression in liver

Background  

Conjugated linoleic acid (CLA) is a collective term for isomers of octadecadienoic acid with conjugated double-bond system. Thus, it was the objective to investigate whether milk composition and metabolic key parameters are affected by adding CLA to the diet of dairy cows in the first four weeks of lactation.     

Predicting grass dry matter intake,milk yield and milk fat and protein yield of spring calving grazing dairy cows during the grazing season

Predicting the grass dry matter intake (GDMI), milk yield (MY) or milk fat and protein yield (milk solids yield (MSY)) of the grazing dairy herd is difficult. Decisions with regard to grazing management are based on guesstimates of the GDMI of the herd, yet GDMI is a critical factor influencing MY and MSY. A data set containing animal, sward, grazing management and concentrate supplementation variables recorded during weeks of GDMI measurement was used to develop multiple regression equations to predict GDMI, MY and MSY. The data set contained data from 245 grazing herds from 10 published studies conducted at Teagasc, Moorepark. A forward stepwise multiple regression technique was used to develop the multiple regression equations for each of the dependent variables (GDMI, MY, MSY) for three periods during the grazing season: spring (SP; 5 March to 30 April), summer (SU; 1 May to 31 July) and autumn (AU; 1 August to 31 October). The equations generated highlighted the importance of different variables associated with GDMI, MY and MSY during the grazing season. Peak MY was associated with an increase in GDMI, MY and MSY during the grazing season with the exception of GDMI in SU when BW accounted for more of the variation. A higher body condition score (BCS) at calving was associated with a lower GDMI in SP and SU and a lower MY and MSY in all periods. A higher BCS was associated with a higher GDMI in SP and SU, a higher MY in SU and AU and a higher MSY in all periods. The pre-grazing herbage mass of the sward (PGHM) above 4 cm was associated with a quadratic effect on GDMI in SP, on MY in SP and SU and on MSY in SU. An increase in daily herbage allowance (DHA) above 4 cm was associated with an increase in GDMI in AU, an increase in MY in SU and AU and MSY in AU. Supplementing grazing dairy cows with concentrate reduced GDMI and increased MY and MSY in all periods. The equations generated can be used by the Irish dairy industry during the grazing season to predict the GDMI, MY and MSY of grazing dairy herds.     

Rumen-protected conjugated linoleic acid supplementation to dairy cows in late pregnancy and early lactation: effects on milk composition,milk yield,blood metabolites and gene expression in liver

After 27 years with no detected cases, an outbreak of anthrax occurred in a beef cattle herd in the south of Sweden. The outbreak was unusual as it occurred in winter, in animals not exposed to meat-and-bone meal, in a non-endemic country. The affected herd consisted of 90 animals, including calves and young stock. The animals were kept in a barn on deep straw bedding and fed only roughage. Seven animals died during 10 days, with no typical previous clinical signs except fever. The carcasses were reportedly normal in appearance, particularly as regards rigor mortis, bleeding and coagulation of the blood. Subsequently, three more animals died and anthrax was suspected at necropsy and confirmed by culture and PCR on blood samples. The isolated strain was susceptible to tetracycline, ciprofloxacin and ampicillin. Subtyping by MLVA showed the strain to cluster with isolates in the A lineage of Bacillus anthracis. Environmental samples from the holding were all negative except for two soil samples taken from a spot where infected carcasses had been kept until they were picked up for transport. The most likely source of the infection was concluded to be contaminated roughage, although this could not be substantiated by laboratory analysis. The suspected feed was mixed with soil and dust and originated from fields where flooding occurred the previous year, followed by a dry summer with a very low water level in the river allowing for the harvesting on soil usually not exposed. In the early 1900s, animal carcasses are said to have been dumped in this river during anthrax outbreaks and it is most likely that some anthrax spores could remain in the area. The case indicates that untypical cases in non-endemic areas may be missed to a larger extent than previously thought. Field tests allowing a preliminary risk assessment of animal carcasses would be helpful for increased sensitivity of detection and prevention of further exposure to the causative agent.     

Effects of flaxseed supplementation on milk production,milk fatty acid composition and nutrient utilization by lactating dairy cows

Twelve multiparous Holstein cows at 72 ± 20 days in milk were used in a switch-back design with 14-d periods to determine the effect of replacing barley grain into a dairy total mixed ration with micronized or raw flaxseed on nutrient digestibility, milk yield, milk composition. Total mixed diets were (DM basis) 50% barley silage, 50% concentrate mix mainly rolled barley grain and canola meal. Diets were supplemented with 1 kg raw (RF) or micronized (MF) flaxseed to substitute 1 kg of rolled barley grain (C). Neutral detergent fibre, ADF and CP digestibility of the diets were not significantly affected by supplementation; however, calcium digestibility was reduced by 62% and 46% when raw and micronized flax were fed, respectively. Milk yield (38.3, 39.6, and 38.4 kg/d for diets C, RF and MF, respectively) was similar for all diets. Milk fat (3.50, 3.48, and 3.52%) and protein (3.31, 3.34, and 3.31%) for diets C, RF and MF, respectively, were not affected by treatment diets. Concentrations of c9, t11 conjugated linoleic acid (CLA; 0.51, 0.72 and 0.76 g/100 g fatty acids) in milk fat increased (P < 0.05) similarly among the two flaxseed supplemented diets. The RF and MF diets significantly increased the C18:1, C18:1 trans-11, C18:2 cis-9, cis-12 and C18:3 in milk fat however, C12:0, C14:0 and C16:0 were significantly reduced compared with control. Replacing barley grain with flaxseed in the diet of lactating cows increased the beneficial fatty acids in milk without depressing nutrient digestibility. Micronization of flaxseed did not reveal any advantage over raw flaxseed.     

Breed of cow and herd productivity affect milk nutrient recovery in curd,and cheese yield,efficiency and daily production

Little is known about cheese-making efficiency at the individual cow level, so our objective was to study the effects of herd productivity, individual herd within productivity class and breed of cow within herd by producing, then analyzing, 508 model cheeses from the milk of 508 cows of six different breeds reared in 41 multi-breed herds classified into two productivity classes (high v. low). For each cow we obtained six milk composition traits; four milk nutrient (fat, protein, solids and energy) recovery traits (REC) in curd; three actual % cheese yield traits (%CY); two theoretical %CYs (fresh cheese and cheese solids) calculated from milk composition; two overall cheese-making efficiencies (% ratio of actual to theoretical %CYs); daily milk yield (dMY); and three actual daily cheese yield traits (dCY). The aforementioned phenotypes were analyzed using a mixed model which included the fixed effects of herd productivity, parity, days in milk (DIM) and breed; the random effects were the water bath, vat, herd and residual. Cows reared in high-productivity herds yielded more milk with higher nutrient contents and more cheese per day, had greater theoretical %CY, and lower cheese-making efficiency than low-productivity herds, but there were no differences between them in terms of REC traits. Individual herd within productivity class was an intermediate source of total variation in REC, %CY and efficiency traits (10.0% to 17.2%), and a major source of variation in milk yield and dCY traits (43.1% to 46.3%). Parity of cows was an important source of variation for productivity traits, whereas DIM affected almost all traits. Breed within herd greatly affected all traits. Holsteins produced more milk, but Brown Swiss cows produced milk with higher actual and theoretical %CYs and cheese-making efficiency, so that the two large-framed breeds had the same dCY. Compared with the two large-framed breeds, the small Jersey cows produced much less milk, but with greater actual and theoretical %CYs, similar efficiencies and a slightly lower dCY. Compared with the average of the specialized dairy breeds, the three dual-purpose breeds (Simmental and the local Rendena and Alpine Grey) had, on average, similar dMY, lower actual and theoretical %CY, similar fat and protein REC, and slightly greater cheese-making efficiency.     

Using pseudo amino acid composition to predict protein subcellular location: Approached with Lyapunov index, Bessel function, and Chebyshev filter

Summary. With the avalanche of new protein sequences we are facing in the post-genomic era, it is vitally important to develop an automated method for fast and accurately determining the subcellular location of uncharacterized proteins. In this article, based on the concept of pseudo amino acid composition (Chou, K.C. Proteins: Structure, Function, and Genetics, 2001, 43: 246–255), three pseudo amino acid components are introduced via Lyapunov index, Bessel function, Chebyshev filter that can be more efficiently used to deal with the chaos and complexity in protein sequences, leading to a higher success rate in predicting protein subcellular location.