Effects of the Flying Start on Estimated Short Sprint Profiles Using Timing Gates.

Short sprints are predominantly assessed using timing gates and analyzed through parameters of the mono-exponential equation, including estimated maximal sprinting speed (MSS) and relative acceleration (TAU), derived maximum acceleration (MAC), and relative propulsive maximal power (PMAX), further referred to as the No Correction model. However, the frequently recommended flying start technique introduces a bias during parameter estimation. To correct this, two additional models (Estimated TC and Estimated FD) were proposed. To estimate model precision and sensitivity to detect the change, 31 basketball players executed multiple 30 m sprints. Athlete performance was simultaneously measured by a laser gun and timing gates positioned at 5, 10, 20, and 30 m. Short sprint parameters were estimated using a laser gun, representing the criterion measure, and five different timing gate models, representing the practical measures. Only the MSS parameter demonstrated a high agreement between the laser gun and timing gate models, using the percent mean absolute difference (%MAD) estimator (%MAD < 10%). The MSS parameter also showed the highest sensitivity, using the minimum detectable change estimator (%MDC95), with an estimated %MDC95 < 17%. Interestingly, sensitivity was the highest for the No Correction model (%MDC95 < 7%). All other parameters and models demonstrated an unsatisfying level of sensitivity. Thus, sports practitioners should be cautious when using timing gates to estimate maximum acceleration indices and changes in their respective levels.

Mechanisms of Hamstring Injury in Professional Soccer Players: Video Analysis and Magnetic Resonance Imaging Findings.

OBJECTIVE: To describe the injury mechanisms and magnetic resonance imaging (MRI) findings in acute hamstring injuries of male soccer players using a systematic video analysis. DESIGN: Descriptive case series study of consecutive acute hamstring injuries from September 2017 to January 2022. SETTING: Two specialized sports medicine hospitals. PARTICIPANTS: Professional male soccer players aged between 18 and 40 years, referred for injury assessment within 7 days after an acute hamstring injury, with an available video footage of the injury and positive finding on MRI. INDEPENDENT VARIABLES: Hamstring injury mechanisms (specific scoring based on standardized models) in relation to hamstring muscle injury MRI findings. MAIN OUTCOME MEASURES: Hamstring injury mechanism (playing situation, player/opponent behavior, movement, and biomechanical body positions) and MRI injury location. RESULTS: Fourteen videos of acute hamstring injuries in 13 professional male soccer players were analyzed. Three different injury mechanisms were seen: mixed-type (both sprint-related and stretch-related, 43%), stretch-type (36%), and sprint-type (21%). Most common actions during injury moments were change of direction (29%), kicking (29%), and running (21%). Most injuries occurred at high or very high horizontal speed (71%) and affected isolated proximal biceps femoris (BF) (36%). Most frequent body positions at defined injury moments were neutral trunk (43%), hip flexion 45-90 degrees (57%), and knee flexion <45 degrees (93%). Magnetic resonance imaging findings showed that 79% were isolated single-tendon injuries. CONCLUSIONS: According to a video analysis, most hamstring injuries in soccer occur during high-speed movements. Physicians should suspect proximal and isolated single-tendon-most often BF-hamstring injury, if represented injury mechanisms are seen during game play. In addition to sprinting and stretching, also mixed-type injury mechanisms occur.

Functional Characterization of the Nemertide α Family of Peptide Toxins.

Peptide toxins find use in medicine, biotechnology, and agriculture. They are exploited as pharmaceutical tools, particularly for the investigation of ion channels. Here, we report the synthesis and activity of a novel family of peptide toxins: the cystine-knotted α nemertides. Following the prototypic α-1 and -2 (1 and 2), six more nemertides were discovered by mining of available nemertean transcriptomes. Here, we describe their synthesis using solid phase peptide chemistry and their oxidative folding by using an improved protocol. Nemertides α-2 to α-7 (2-7) were produced to characterize their effect on voltage-gated sodium channels (Blatella germanica BgNaV1 and mammalian NaVs1.1-1.8). In addition, ion channel activities were matched to in vivo tests using an Artemia microwell assay. Although nemertides demonstrate high sequence similarity, they display variability in activity on the tested NaVs. The nemertides are all highly toxic to Artemia, with EC50 values in the sub-low micromolar range, and all manifest preference for the insect BgNaV1 channel. Structure-activity relationship analysis revealed key residues for NaV-subtype selectivity. Combined with low EC50 values (e.g., NaV1.1: 7.9 nM (α-6); NaV1.3: 9.4 nM (α-5); NaV1.4: 14.6 nM (α-4)) this underscores the potential utility of α-nemertides for rational optimization to improve selectivity.

Functionality of primary hepatic non-parenchymal cells in a 3D spheroid model and contribution to acetaminophen hepatotoxicity.

In addition to hepatocytes, the liver comprises a host of specialised non-parenchymal cells which are important to consider in the development of in vitro models which are both physiologically and toxicologically relevant. We have characterized a 3D co-culture system comprising primary human hepatocytes (PHH) and non-parenchymal cells (NPC) and applied it to the investigation of acetaminophen-induced toxicity. Firstly, we titrated ratios of PHH:NPC and confirmed the presence of functional NPCs via both immunohistochemistry and activation with both LPS and TGF-ß. Based on these data we selected a ratio of 2:1 PHH:NPC for further studies. We observed that spheroids supplemented with NPCs were protected against acetaminophen (APAP) toxicity as determined by ATP (up to threefold difference in EC50 at day 14 compared to hepatocytes alone) and glutathione depletion, as well as miR-122 release. APAP metabolism was also altered in the presence of NPCs, with significantly lower levels of APAP-GSH detected. Expression of several CYP450 enzymes involved in the bioactivation of APAP was also lower in NPC-containing spheroids. Spheroids containing NPCs also expressed higher levels of miRNAs which have been implicated in APAP-induced hepatotoxicity, including miR-382 and miR-155 which have potential roles in liver regeneration and inflammation, respectively. These data indicate that the interaction between hepatocytes and NPCs can have significant metabolic and toxicological consequences important for the correct elucidation of hepatic safety mechanisms.

Synthesis of di-docosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate in unlabelled and C-13 labelled forms for use as a biomarker of drug induced phospholipidosis.

Di-docosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate (BMP) has been identified as a promising biomarker for drug-induced phospholipidosis (DIPL). Both unlabelled and stable isotope labelled versions of BMP were desired for use as internal standards. Isopropylideneglycerol was converted to 4-methoxyphenyldiphenylmethyl-3-PMB-glycerol in three steps. Initially, the 2-postion of the glycerol was protected as a t-butyldiphenylsilyl ether, which proved to be a mistake; deprotection of the ether resulted in the decomposition of the compound. A switch to a t-butyldimethylsilyl ether protecting group resulted in an intermediate that could be deprotected to the alcohol to give the target compound after salt exchange. The same procedure was used to prepare [13 C6 ]BMP from [13 C3 ]glycerol.

Naloxegol, an opioid antagonist with reduced CNS penetration: Mode-of-action and human relevance for rat testicular tumours.

Naloxegol is an opioid antagonist which has been developed for the treatment of patients with opioid induced constipation. In the nonclinical safety program naloxegol was shown to have a very benign toxicity profile. In the rat, but not the mouse, 2-year carcinogenicity study a change in tumour pattern with an increase in testicular Leydig cell tumours (LCT) was observed after dosing at high (supra-pharmacological) concentrations. To establish the basis of the increase in LCT and to assess its potential relevance to humans, studies to exclude and potentially identify mode-of-action (MoA) were performed. A genotoxic mechanism was ruled out following negative results in the Ames, mouse lymphoma, and micronucleus assays. An effect on androgen metabolism was excluded since the treatment of rats with naloxegol for 14days did not result in any induction of CYP protein levels. It was demonstrated that administration of centrally restricted opioid antagonists naloxegol or methylnaltrexone at high doses induced an increase in LH release with no clear increase in testosterone, in contrast to the centrally acting opioid antagonist naloxone, which showed marked increases in both LH and testosterone. LCT due to increased LH stimulation is common in rats but not documented in humans. Collectively, the lack of genotoxicity signal, the lack of androgen effect, the increase in LH secretion in rats, which is no considered to be relevant for LCT formation in humans, and high margins to clinical exposures, the observed increase in LCT in the rat is not expected to be clinically relevant.

The Bacterial (Vibrio alginolyticus) Production of Tetrodotoxin in the Ribbon Worm Lineus longissimus-Just a False Positive?

We test previous claims that the bacteria Vibrio alginolyticus produces tetrodotoxin (TTX) when living in symbiosis with the nemertean Lineus longissimus by a setup with bacteria cultivation for TTX production. Toxicity experiments on the shore crab, Carcinus maenas, demonstrated the presence of a paralytic toxin, but evidence from LC-MS and electrophysiological measurements of voltage-gated sodium channel-dependent nerve conductance in male Wistar rat tissue showed conclusively that this effect did not originate from TTX. However, a compound of similar molecular weight was found, albeit apparently non-toxic, and with different LC retention time and MS/MS fragmentation pattern than those of TTX. We conclude that C. maenas paralysis and death likely emanate from a compound <5 kDa, and via a different mechanism of action than that of TTX. The similarity in mass between TTX and the Vibrio-produced low-molecular-weight, non-toxic compound invokes that thorough analysis is required when assessing TTX production. Based on our findings, we suggest that re-examination of some published claims of TTX production may be warranted.

Molecular motor propelled filaments reveal light-guiding in nanowire arrays for enhanced biosensing.

Semiconductor nanowire arrays offer significant potential for biosensing applications with optical read-out due to their high surface area and due to the unique optical properties of one-dimensional materials. A challenge for optical read-out of analyte-binding to the nanowires is the need to efficiently collect and detect light from a three-dimensional volume. Here we show that light from fluorophores attached along several µm long vertical Al2O3 coated gallium phosphide nanowires couples into the wires, is guided along them and emitted at the tip. This enables effective collection of light emitted by fluorescent analytes located at different focal planes along the nanowire. We unequivocally demonstrate the light-guiding effect using a novel method whereby the changes in emitted fluorescence intensity are observed when fluorescent cytoskeletal filaments are propelled by molecular motors along the wires. The findings are discussed in relation to nanobiosensor developments, other nanotechnological applications, and fundamental studies of motor function.

Dropout revisited: patient- and therapist-initiated discontinuation of psychotherapy as a function of organizational instability.

OBJECTIVE: To explore the association between the stability or instability of services' organizational structure and patient- and therapist-initiated discontinuation of therapy in routine mental health. METHOD: Three groups, comprising altogether 750 cases in routine mental health care in eight different clinics, were included: cases with patient-initiated discontinuation, therapist-initiated discontinuation, and patients remaining in treatment. Multilevel multinomial regression was used to estimate three models: An initial, unconditional intercept-only model, another one including patient variables, and a final model with significant patient and therapist variables including the organizational stability of the therapists' clinic. RESULTS: High between-therapist variability was noted. Odds ratios and significance tests indicated a strong association of organizational instability with patient-initiated premature termination in particular. CONCLUSIONS: The question of how organizational factors influence the treatment results needs further research. Future studies have to be designed in ways that permit clinically meaningful subdivision of the patients' and the therapists' decisions for premature termination.

Peptide Toxins from Antarctica: The Nemertean Predator and Scavenger Parborlasia corrugatus (McIntosh, 1876).

Peptide toxins from marine invertebrates have found use as drugs and in biotechnological applications. Many marine habitats, however, remain underexplored for natural products, and the Southern Ocean is among them. Here, we report toxins from one of the top predators in Antarctic waters: the nemertean worm Parborlasia corrugatus (McIntosh, 1876). Transcriptome mining revealed a total of ten putative toxins with a cysteine pattern similar to that of alpha nemertides, four nemertide-beta-type sequences, and two novel full-length parborlysins. Nemertean worms express toxins in the epidermal mucus. Here, the expression was determined by liquid chromatography combined with mass spectrometry. The findings include a new type of nemertide, 8750 Da, containing eight cysteines. In addition, we report the presence of six cysteine-containing peptides. The toxicity of tissue extracts and mucus fractions was tested in an Artemia assay. Notably, significant activity was observed both in tissue and the high-molecular-weight mucus fraction, as well as in a parborlysin fraction. Membrane permeabilization experiments display the membranolytic activity of some peptides, most prominently the parborlysin fraction, with an estimated EC50 of 70 nM.

Amide hydrolysis of a novel chemical series of microsomal prostaglandin E synthase-1 inhibitors induces kidney toxicity in the rat.

A novel microsomal prostaglandin E synthase 1 (mPGES-1) inhibitor induced kidney injury at exposures representing less than 4 times the anticipated efficacious exposure in man during a 7-day toxicity study in rats. The findings consisted mainly of tubular lesions and the presence of crystalline material and increases in plasma urea and creatinine. In vitro and in vivo metabolic profiling generated a working hypothesis that a bis-sulfonamide metabolite (determined M1) formed by amide hydrolysis caused this toxicity. To test this hypothesis, rats were subjected to a 7-day study and were administered the suspected metabolite and two low-potency mPGES-1 inhibitor analogs, where amide hydrolysis was undetectable in rat hepatocyte experiments. The results suggested that compounds with a reduced propensity to undergo amide hydrolysis, thus having less ability to form M1, reduced the risk of inducing kidney toxicity. Rats treated with M1 alone showed no histopathologic change in the kidney, which was likely related to underexposure to M1. To circumvent rat kidney toxicity, we identified a potent mPGES-1 inhibitor with a low propensity for amide hydrolysis and superior rat pharmacokinetic properties. A subsequent 14-day rat toxicity study showed that this compound was associated with kidney toxicity at 42, but not 21, times the anticipated efficacious exposure in humans. In conclusion, by including metabolic profiling and exploratory rat toxicity studies, a new and active mPGES-1 inhibitor with improved margins to chemically induced kidney toxicity in rats has been identified.

Cancer odor in the blood of ovarian cancer patients: a retrospective study of detection by dogs during treatment, 3 and 6 months afterward.

BACKGROUND: In recent decades it has been noted that trained dogs can detect specific odor molecules emitted by cancer cells. We have shown that the same odor can also be detected in the patient's blood with high sensitivity and specificity by trained dogs. In the present study, we examined how the ability of dogs to detect this smell was affected by treatment to reduce tumor burden, including surgery and five courses of chemotherapy. METHODS: In Series I, one drop of plasma from each of 42 ovarian cancer patients (taken between the fifth and sixth courses of chemotherapy) and 210 samples from healthy controls were examined by two trained dogs. All 42 patients in Series I had clinical complete responses, all except two had normal CA-125 values and all were declared healthy after primary treatment. In Series II, the dogs examined blood taken from a new subset of 10 patients at 3 and 6 months after the last (sixth) course of chemotherapy. RESULTS: In Series I, the dogs showed high sensitivity (97%) and specificity (99%), for detecting viable cancer cells or molecular cancer markers in the patients' plasma. Indeed, 29 of 42 patients died within 5 years. In Series II, the dogs indicated positive samples from three of the 10 patients at both the 3- and 6-month follow-up. All three patients had recurrences, and two died 3-4 years after the end of treatment. This was one of the most important findings of this study. Seven patients were still alive in January 2013. CONCLUSIONS: Although our study was based on a limited number of selected patients, it clearly suggests that canine detection gave us a very good assessment of the prognosis of the study patients. Being able to detect a marker based on the specific cancer odor in the blood would enhance primary diagnosis and enable earlier relapse diagnosis, consequently increasing survival.

Systems pharmacology modeling of drug-induced modulation of thyroid hormones in dogs and translation to human.

PURPOSE: To develop a systems pharmacology model based on hormone physiology and pharmacokinetic-pharmacodynamic concepts describing the impact of thyroperoxidase (TPO) inhibition on thyroid hormone homeostasis in the dog and to predict drug-induced changes in thyroid hormones in humans. METHODS: A population model was developed based on a simultaneous analysis of concentration-time data of T4, T3 and TSH in dogs following once daily oral dosing for up to 6-months of a myeloperoxidase inhibitor (MPO-IN1) with TPO inhibiting properties. The model consisted of linked turnover compartments for T4, T3 and TSH including a negative feedback from T4 on TSH concentrations. RESULTS: The model could well describe the concentration-time profiles of thyroid hormones in dog. Successful model validation was performed by predicting the hormone concentrations during 1-month administration of MPO-IN2 based on its in vitro dog TPO inhibition potency. Using human thyroid hormone turnover rates and TPO inhibitory potency, the human T4 and TSH concentrations upon MPO-IN1 treatment were predicted well. CONCLUSIONS: The model provides a scientific framework for the prediction of drug induced effects on plasma thyroid hormones concentrations in humans via TPO inhibition based on results obtained in in vitro and animal studies.

Scientific and regulatory policy committee (SRPC) paper: assessment of circulating hormones in nonclinical toxicity studies III. female reproductive hormones.

Hormonally mediated effects on the female reproductive system may manifest as pathologic changes of endocrine-responsive organs and altered reproductive function. Identification of these effects requires proper assessment, which may include investigative studies to profile female reproductive hormones. Here, we briefly describe normal hormonal patterns across the estrous or menstrual cycle and provide general guidance on measuring female reproductive hormones and characterizing hormonal disturbances in nonclinical toxicity studies. Although species used in standard toxicity studies share basic features of reproductive endocrinology, there are important species differences that affect both study design and interpretation of results. Diagnosing female reproductive hormone disturbances can be complicated by many factors, including estrous/menstrual cyclicity, diurnal variation, and age- and stress-related factors. Thus, female reproductive hormonal measurements should not generally be included in first-tier toxicity studies of standard design with groups of unsynchronized intact female animals. Rather, appropriately designed and statistically powered investigative studies are recommended in order to properly identify ovarian and/or pituitary hormone changes and bridge these effects to mechanistic evaluations and safety assessments. This article is intended to provide general considerations and approaches for these types of targeted studies.

Differences in Peak Knee Flexor Force between Eccentric-Only and Combined Eccentric-Concentric Nordic Hamstring Exercise.

In many sports, the hamstring strain injury is a common injury. There is evidence that the Nordic hamstring exercise (NHE), a knee flexor exercise, can reduce hamstring injury risk in athletes. In research on hamstring injury prevention, eccentric-only NHE is typically performed, whereas in sports, it is relatively common for athletes to perform NHE eccentrically-concentrically. Further, NHE strength is generally assessed by measuring knee flexor force through an ankle brace, attached atop of a load cell. An alternative method might be to assess knee flexor force about the knee joint using a force plate. The aim of the study was to investigate differences in peak knee flexor force between eccentric-only and combined eccentric-concentric NHE. The purpose was also to determine the correlation between hamstring force measured at the ankle using a load cell (current gold standard) and force assessed about the knee joint using a force plate during NHE. Fifteen junior and senior elite soccer and track and field athletes (3 women and 12 men aged 17-27 years) performed eccentric NHE (ENHE) in which they leaned forward as far as possible until breakpoint and eccentric-concentric NHE (ECNHE) where they returned to the starting position. A linear encoder measured the position at which peak force occurred during the NHEs. Force assessed at the ankle differed significantly (678 vs. 600 N, p < 0.05), whereas force about the knee joint did not (640 vs. 607 N, p > 0.05) between ENHE and ECNHE (12 and 5% difference, respectively). The forward distance achieved by the participants in cm at breakpoint for ENHE was 37% higher than at the coupling phase for ECNHE (74 vs. 54 cm, p < 0.001). Very strong significant (p < 0.01) correlations were noted between peak force assessed at the ankle and about the knee joint for ENHE and ECNHE, r = 0.96 and r = 0.99, respectively. Our results suggest that ECNHE, where peak knee flexor force was reached with 37% less forward movement, may complement ENHE, i.e., during hamstring injury rehabilitation, where a position of great knee extension may not be well tolerated by the athlete. Further, assessing knee flexor force about the knee joint using a force plate may provide an alternative to measuring force at the ankle using a load cell when testing NHE strength.

Speed Matters in Nordic Hamstring Exercise: Higher Peak Knee Flexor Force during Fast Stretch-Shortening Variant Compared to Standard Slow Eccentric Execution in Elite Athletes.

Hamstring strain injuries are prevalent in many sports. Research has demonstrated that the Nordic hamstring exercise (NHE), a knee-dominant exercise addressing the posterior chain muscles, can aid in reducing the risk of hamstring injuries in athletes. However, most research on hamstring injury prevention has focused on performing the eccentric version of the NHE (NHEECC). In contrast, in sports, it is quite frequent for athletes to use an eccentric-concentric version of the NHE. Additionally, eccentric NHE is typically performed using a slow, controlled tempo. The effect of a fast stretch-shortening cycle NHE (NHESSC) compared to standard slow NHEECC on peak knee flexor force has not been investigated. The aim of the study was therefore to investigate fast NHESSC vs. standard slow NHEECC. Our hypothesis posited that peak knee flexor force would be greater for fast NHESSC compared with standard slow NHEECC. The study involved 22 elite athletes (actively competing in both national and international events) consisting of female (n = 10) and male (n = 7) track and field athletes and male football players (n = 5), aged 17-31 years. The participants performed maximum trials of slow NHEECC and fast NHESSC repetitions in which measurement of bilateral peak knee flexor force was conducted at the ankle with the use of a load cell. During the NHEs, a linear encoder was used to measure both the position where the peak knee flexor force was recorded and the average eccentric velocity. SSC contributed to an enhanced NHE performance, where bilateral absolute peak knee flexor force was 13% higher for fast NHESSC vs. standard slow NHEECC (822 vs. 726 N, p < 0.01, ES = 0.54). Participants achieved a 32% greater forward distance at the breakpoint stage during NHEECC compared to the coupling phase for NHESSC (54 vs. 41 cm, p < 0.001, ES = 1.37). Eccentric average velocity was more than three times higher for NHESSC compared with NHEECC (0.38 vs. 0.12 m/s, p < 0.001, ES = 3.25). The key findings of this study were that SSC contributed to an enhanced NHE performance, where absolute peak knee flexor force was 13% greater for fast NHESSC compared to standard slow NHEECC. The fast NHESSC could therefore be an interesting alternative to the standard slow NHEECC execution, as it may offer potential advantages for sprint performance, as well as hamstring injury prevention and rehabilitation.

Sprint skating profile of competitive female bandy players: An analysis of positional and playing-level differences.

There is no research examining female bandy players, which creates a gap of knowledge of female skating performance and its determinants with male skating performance, not only in bandy but also in exercise science in general. Therefore, the aim of this explorative study was to investigate position and playing-level differences in the sprint skating performance and anthropometrics of 74 elite female bandy players (age: 18.9 ± 4.1 years; height: 1.67 ± 0.06 m; body mass: 63.2 ± 7.4 kg). Participants were categorised according to playing level (26 elite and 48 junior elite players) and position (22 defenders, 35 midfielders, and 17 forwards). They were tested on their anthropometric characteristics and sprint linear skating profile over 80 m with the split times measured at 10, 20, 40, 50, 60, 70, and 80 m to calculate the average velocities between these different 10 m intervals. Results revealed that elite players had more training experience, were heavier, could accelerate faster, and reached a higher maximal velocity than the junior elite players (9.52 ± 0.37 vs 8.84 ± 0.40 m/s, respectively). In general, defenders were heavier than forwards, and the elite forwards accelerated faster in the first 10 m than the midfielders (p = 0.041). In summary, playing level and position, body mass, and training experience modulated skating sprint performance. The findings suggest that female junior-level players should spend more time developing skating sprint and acceleration abilities to meet the specific demands of playing at the elite level. Moreover, the coaches and professionals who work with female bandy players should be aware that the development of acceleration ability is more important for forwards.

Sprint skating profile of competitive male bandy players: determination of positional differences and playing level.

This study aimed to compare sprint skating profile characteristics of the different playing positions of junior and senior bandy players. In total, 111 male national-level bandy players (age: 20.7 ± 5.0 years, height: 1.80 ± 0.05 m, body mass: 76.4 ± 0.4 kg, training experience: 13.8 ± 5.0 yrs) were tested on their sprint skating profile over 80 m. The main findings were that no differences between positions were found in sprint skating performance (speed and acceleration), but that elite players were in general heavier (p < 0.05) than junior players (80.0 ± 7.1 vs. 73.1 ± 8.1 kg), they could accelerate faster (2.96 ± 0.22 vs. 2.81 ± 0.28 m/s2), and they reached a higher velocity (10.83 ± 0.37 vs. 10.24 ± 0.42 m/s) earlier over 80 m than the junior players. This implies that junior level players should spend more time in power and sprint training to meet the specific demands of playing at a higher, elite level.

The α-defensin salt-bridge induces backbone stability to facilitate folding and confer proteolytic resistance.

Salt-bridge interactions between acidic and basic amino acids contribute to the structural stability of proteins and to protein-protein interactions. A conserved salt-bridge is a canonical feature of the α-defensin antimicrobial peptide family, but the role of this common structural element has not been fully elucidated. We have investigated mouse Paneth cell α-defensincryptdin-4 (Crp4) and peptide variants with mutations at Arg7 or Glu15 residue positions to disrupt the salt-bridge and assess the consequences on Crp4 structure, function, and stability. NMR analyses showed that both (R7G)-Crp4 and (E15G)-Crp4 adopt native-like structures, evidence of fold plasticity that allows peptides to reshuffle side chains and stabilize the structure in the absence of the salt-bridge. In contrast, introduction of a large hydrophobic side chain at position 15, as in (E15L)-Crp4 cannot be accommodated in the context of the Crp4 primary structure. Regardless of which side of the salt-bridge was mutated, salt-bridge variants retained bactericidal peptide activity with differential microbicidal effects against certain bacterial cell targets, confirming that the salt-bridge does not determine bactericidal activity per se. The increased structural flexibility induced by salt-bridge disruption enhanced peptide sensitivity to proteolysis. Although sensitivity to proteolysis by MMP7 was unaffected by most Arg(7) and Glu(150 substitutions, every salt-bridge variant was degraded extensively by trypsin. Moreover, the salt-bridge facilitates adoption of the characteristic α-defensin fold as shown by the impaired in vitro refolding of (E15D)-proCrp4, the most conservative salt-bridge disrupting replacement. In Crp4, therefore, the canonical α-defensin salt-bridge facilitates adoption of the characteristic α-defensin fold, which decreases structural flexibility and confers resistance todegradation by proteinases.

A review of the incidence and coincidence of uterine and mammary tumors in Wistar and Sprague-Dawley rats based on the RITA database and the role of prolactin.

Wistar rats are frequently selected for use in carcinogenicity studies because of their advantageous survival rate, which is more favorable than other strains such as the Sprague-Dawley (SD) strain. Uterine and mammary tumors are relatively common spontaneous neoplasms of both strains. We examined the incidence and coincidence of uterine tumors and mammary tumors in control animals of both strains within the RITA database. There was a strong inverse relationship between these tumor types in Wistar rats (p < .001). A less strong relationship was present in SD rats (p = .057). This association is likely to be related to prolactin. A short review of the role of prolactin in rats is given. These results are also discussed in the background of nonspecific toxicity at high dose levels in carcinogenicity studies above MTD levels resulting in reduction in body weights of >10%.