Efficacy of ADIPOR1 and ADIPOR2 peptide-agonist AdipoRon in preventing contracture in a rabbit model of arthrofibrosis.

AdipoRon is an adiponectin receptor 1, 2 (ADIPOR1 and ADIPOR2) agonist with potential antifibrotic effects. Whether AdipoRon can mitigate joint stiffness in a rabbit model of arthrofibrosis is unknown. We examined the efficacy of intravenous (IV) AdipoRon at mitigating contracture in a rabbit model of knee arthrofibrosis. Fifty-six female New Zealand White rabbits were divided into three dosing groups: vehicle (dimethyl sulfoxide, DMSO), 2.5 mg/kg AdipoRon, and 5 mg/kg AdipoRon. AdipoRon, in DMSO, was administered IV preoperatively and for 5 days postoperatively (30 rabbits, Aim 1). AdipoRon was again dosed similarly after Kirschner wire (K-wire) removal at 8 weeks (26 rabbits; Aim 2). The primary outcome of joint passive extension angle (PEA,°) was measured at 8, 10, 12, 16, and 24 weeks following index surgery. At 24 weeks, rabbits were euthanized and limbs were harvested to measure posterior capsular stiffness (N cm/°). In Aim 1, the 5 mg/kg treated rabbits had a significant increase in PEA when compared to controls at 16-week (p < 0.05). In Aim 2, the 5 mg/kg treated rabbits had a significant increase in PEA when compared to controls at 10-week (p < 0.05). In both aims, no significant differences were observed at later time points. Capsular stiffness was no different in any group. We are the first to report the efficacy of IV AdipoRon in a rabbit model of arthrofibrosis. We identified a significant dose-dependent decrease in joint PEA at early time points; however, no differences were observed between groups at later time points. Clinical Significance: The present investigation provided the first assessment of AdipoRon's efficacy in mitigating knee stiffness in the current gold standard rabbit model of arthrofibrosis. Results of this investigation provided further evidence as to the potential role of AdipoRon as a preventative for arthrofibrosis in large mammals.

AdipoRon reduces TGFß1-mediated collagen deposition in vitro and alleviates knee stiffness in vivo.

Arthrofibrosis, which causes joint motion restrictions, is a common complication following total knee arthroplasty (TKA). Key features associated with arthrofibrosis include myofibroblast activation, knee stiffness, and excessive scar tissue formation. We previously demonstrated that adiponectin levels are suppressed within the knee tissues of patients affected by arthrofibrosis and showed that AdipoRon, an adiponectin receptor agonist, exhibited anti-fibrotic properties in human mesenchymal stem cells. In this study, the therapeutic potential of AdipoRon was evaluated on TGFß1-mediated myofibroblast differentiation of primary human knee fibroblasts and in a mouse model of knee stiffness. Picrosirius red staining revealed that AdipoRon reduced TGFß1-induced collagen deposition in primary knee fibroblasts derived from patients undergoing primary TKA and revision TKA for arthrofibrosis. AdipoRon also reduced mRNA and protein levels of ACTA2, a key myofibroblast marker. RNA-seq analysis corroborated the anti-myofibrogenic effects of AdipoRon. In our knee stiffness mouse model, 6 weeks of knee immobilization, to induce a knee contracture, in conjunction with daily vehicle (DMSO) or AdipoRon (1, 5, and 25 mg/kg) via intraperitoneal injections were well tolerated based on animal behavior and weight measurements. Biomechanical testing demonstrated that passive extension angles (PEAs) of experimental knees were similar between vehicle and AdipoRon treatment groups in mice evaluated immediately following immobilization. Interestingly, relative to vehicle-treated mice, 5 mg/kg AdipoRon therapy improved the PEA of the experimental knees in mice that underwent 4 weeks of knee remobilization following the immobilization and therapy. Together, these studies revealed that AdipoRon may be an effective therapeutic modality for arthrofibrosis.

Comparison of the "Tall and Fall" Versus "Drop and Drive" Pitching Styles: Analysis of Major League Baseball Pitchers During a Single Season.

Background: Previous research has documented the proportion of "tall and fall" (TF) and "drop and drive" (DD) pitching styles among Major League Baseball (MLB) pitchers who underwent ulnar collateral ligament reconstruction (UCLR). The proportion of these 2 styles among all MLB pitchers remains unknown. Purpose: To determine the proportion of the TF and DD pitching styles in all rostered MLB pitchers during a single season as well as the proportion of TF and DD pitchers who sustained an upper extremity (UE) injury and those who underwent UCLR. Study Design: Cross-sectional study; Level of evidence, 3. Methods: Pitcher demographic characteristics from the 2019 MLB season and pitching information were obtained via open-access sources. Two-dimensional video analysis was used to categorize the included pitchers into TF and DD groups. Statistical comparisons and contrasts were made using 2-tailed t tests, chi-square tests, and Pearson correlation analyses as appropriate. Results: Of the 660 MLB rostered pitchers in 2019 (age, 27.39 ± 3.51 years; body mass index, 26.34 ± 2.47 kg/m2; fastball velocity, 150.49 ± 3.99 kph [93.51 ± 2.48 mph]), 412 (62.4%) pitchers used the TF style and 248 (37.6%) pitchers used the DD style. Significantly more UE injuries were seen in the TF group compared with the DD group (112 vs 38 injuries, respectively; P < .001). Twelve pitchers underwent UCLR (TF, 10; DD, 2), representing a 1.8% UCLR rate among all pitchers. This was a second surgery for 2 pitchers, both of whom used the TF pitching style. Significantly more pitchers in the TF group than the DD group had undergone UCLR before 2019 (135 vs 56 pitchers, respectively; P = .005). Conclusion: The results of the present study demonstrated a higher prevalence of both UE injury and prior UCLR in TF pitchers. Further research is needed to explore the potential association between pitching style and UE injury.

The Safety of Adiponectin Receptor Agonist AdipoRon in a Rabbit Model of Arthrofibrosis.

AdipoRon is an adiponectin receptor 1, 2 (ADIPOR1 and ADIPOR2) agonist with numerous reported physiological benefits in murine models of human disease, including a proposed reduction in fibrosis. However, AdipoRon has never been investigated in rabbits, which provide a robust model for orthopedic conditions. We examined the safety of intravenous (IV) AdipoRon in New Zealand White (NZW) female rabbits surgically stressed by a procedure that mimics human arthrofibrosis. Fifteen female NZW rabbits were prospectively studied using increasing AdipoRon doses based on established literature. AdipoRon was dissolved in dimethyl sulfoxide (DMSO), diluted in normal saline, and administered IV preoperatively and for 5 subsequent days postoperatively. The primary outcome was overall toxicity to rabbits, whereas secondary outcomes were change in rabbit weights and hemodynamics and defining acid-base characteristics of the drug formulation. Two rabbits expired during preoperative drug administration at 25 mg/kg. Remaining rabbits received preoperative doses of DMSO (vehicle), 2.5, 5, or 10 mg/kg of AdipoRon without complications. On postoperative day 1, one rabbit sustained a tonic-clonic seizure after their second dose of 10 mg/kg AdipoRon. The remaining 12 rabbits (4 in each group) received six serial doses of vehicle, 2.5, or 5 mg/kg of AdipoRon without adverse effects. All formulations of AdipoRon were within safe physiological pH ranges (4-5). We are the first to report the use of IV AdipoRon in a surgically stressed rabbit model of orthopedic disease. AdipoRon doses of 5 mg/kg or less appear to be well-tolerated in female NZW rabbits. Impact statement We provided the first in vivo toxicity assessment and dose optimization of a new antifibrotic experimental medication, AdipoRon, in a surgically stressed rabbit model of knee arthrofibrosis.

Knowledge and Perceptions of Ulnar Collateral Ligament Injuries in Baseball Players: A Survey of NCAA Head Baseball Coaches.

Background: There is growing concern among sports medicine professionals regarding the increasing prevalence of ulnar collateral ligament (UCL) surgery in baseball players. At this time, it is unclear whether collegiate head baseball coaches possess adequate knowledge of UCL injuries. Purpose/Hypothesis: The purpose of this study was to assess the knowledge and perceptions of UCL injury among National Collegiate Athletic Association (NCAA) head baseball coaches. It was hypothesized that NCAA head baseball coaches would demonstrate misconceptions and knowledge disparities regarding UCL injury. Study Design: Cross-sectional study. Methods: An online 35-question survey was designed to assess NCAA head baseball coaches' knowledge of UCL injury. The survey included questions related to participant characteristics, UCL injury, biometric performance and pitching variables, throwing fatigue, surgical variables/performance outcomes, and collegiate athlete recruitment. A total of 788 NCAA head baseball coaches were asked to participate. Responses were analyzed and reported using descriptive statistics where appropriate. Statistical comparisons and contrasts were made using chi-square and Fisher exact tests. Results: A total of 103 NCAA head baseball coaches participated in the survey, representing a 13.1% response rate. Only 31% of respondents could correctly identify all UCL injury symptoms, and 93% recommended <8 weeks off from overhead throwing during the off-season. Previous elbow injury observation was associated with UCL symptom identification (χ2 = 10.614; P = .005). Regular access to an athletic trainer (P = .015) and regular access to a strength and conditioning coach (P = .004) were both associated with NCAA division status, with Division I programs having the most access. Neither sports medicine physician (P = .656) nor athletic trainer (P = .611) access was associated with the geographic location of the baseball program; however, strength and conditioning coach access was associated with geographic location (χ2 = 6.696; P = .010). Conclusion: This study demonstrates that limited UCL injury knowledge and various misconceptions exist among NCAA baseball head coaches. The majority of responding coaches recommended an amount of time off from overhead throwing during the off-season that may be inadequate and were unable to identify all symptoms associated with UCL injury, representing the most concerning findings of the present study.

Retrospective Analysis of Ulnar Collateral Ligament Reconstructions in Major League Baseball Pitchers: A Comparison of the "Tall and Fall" Versus "Drop and Drive" Pitching Styles.

Background: Previous pilot research has investigated differences in elbow valgus torque between the "tall and fall" (TF) and "drop and drive" (DD) pitching styles. Whether one of these pitching styles is associated with a greater rate of ulnar collateral ligament reconstruction (UCLR) is currently unknown. Purpose: To determine the proportion of Major League Baseball (MLB) pitchers using the TF and DD pitching styles who underwent UCLR over a 10-year period. Study Design: Cross-sectional study; Level of evidence, 3. Methods: The demographic characteristics of pitchers who underwent UCLR between 2007 and 2017 were obtained via the open-source database MLB Player Analysis Tommy John Surgery List. Other information, such as previous UCLR and pitching videos and graphics, was obtained from other open-source databases. A comprehensive, 2-dimensional, kinesiology-based multicomponent definition of each pitching style was formulated and used to categorize the included pitchers into the TF and DD groups. Statistical comparisons and contrasts were made using chi-square and Pearson correlation tests. Results: Included were 223 MLB pitchers (mean ± SD age, 27.5 ± 3.6 years; body mass index [BMI], 27.6 ± 2.2; throwing velocity, 92.9 ± 2.6 mph [149.5 ± 4.2 km/h]) who underwent UCLR between 2007 and 2017. Of these pitchers, 162 were categorized as TF pitchers (72.6%) and 61 as DD pitchers (27.4%). Pitching velocity for injured pitchers was significantly correlated to BMI (P < .001). We found no significant associations of pitching style with year of UCLR (P = .941), BMI (P = .549), age (P = .647), handedness (P = .501), or average pitch velocity (P = .921). Conclusion: The study findings demonstrated that a higher proportion of UCL-injured MLB pitchers (72.6%) used the TF pitching style. Further research is needed to explore the potential association between pitching style and UCL injury.

Differences in Shoulder Internal Rotation Strength Between Baseball Players With Ulnar Collateral Ligament Reconstruction and Healthy Controls.

BACKGROUND: Studies have indicated decreased shoulder internal rotation (IR) and external rotation (ER) strength in the throwing limb of baseball players after ulnar collateral ligament injury. There is limited evidence on the recovery of shoulder rotation strength after primary ulnar collateral ligament reconstruction (UCLR). HYPOTHESIS: At the time of return to throwing, baseball players who underwent UCLR would demonstrate decreased IR and ER shoulder strength in the throwing arm as compared with healthy baseball players. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: Male competitive high school and collegiate baseball athletes participated in this study. Athletes who underwent UCLR were compared with healthy controls who were matched by age, height, weight, and position. Bilateral isometric shoulder ER and IR strength was measured using a handheld dynamometer for all participants at the time of initial evaluation (UCLR group) and throughout the course of a season (healthy group). Independent t tests were run to calculate mean differences in ER and IR shoulder strength between the groups, with significance set at P < .05. RESULTS: A total of 86 baseball athletes participated in this study (43 UCLR group, 43 healthy group). At the time of return to throwing (mean ± SD, 194 ± 30 days postoperatively), the 2 groups demonstrated no significant differences in nonthrowing arm ER or IR strength (P = .143 and .994, respectively). No significant difference was found between groups for throwing arm ER strength (P = .921); however, the UCLR group demonstrated significantly less throwing arm IR strength than the healthy group (144.2 ± 27.8 vs 157.6 ± 27.1 N; P = .023). CONCLUSION: The results of this study demonstrate that throwing arm rotator cuff strength may not fully recover before the initiation of a return-to-throwing program after UCLR. These data provide a potential framework for clinicians to assist in the management and exercise prescription of the baseball athlete after UCLR and before medical release and the initiation of a return-to-throwing program.

The epigenetic reader Brd4 is required for osteoblast differentiation.

Transcription networks and epigenetic mechanisms including DNA methylation, histone modifications, and noncoding RNAs control lineage commitment of multipotent mesenchymal progenitor cells. Proteins that read, write, and erase histone tail modifications curate and interpret the highly intricate histone code. Epigenetic reader proteins that recognize and bind histone marks provide a crucial link between histone modifications and their downstream biological effects. Here, we investigate the role of bromodomain-containing (BRD) proteins, which recognize acetylated histones, during osteogenic differentiation. Using RNA-sequencing (RNA-seq) analysis, we screened for BRD proteins (n = 40) that are robustly expressed in MC3T3 osteoblasts. We focused functional follow-up studies on Brd2 and Brd4 which are highly expressed in MC3T3 preosteoblasts and represent "bromodomain and extra terminal domain" (BET) proteins that are sensitive to pharmacological agents (BET inhibitors). We show that small interfering RNA depletion of Brd4 has stronger inhibitory effects on osteoblast differentiation than Brd2 loss as measured by osteoblast-related gene expression, extracellular matrix deposition, and alkaline phosphatase activity. Similar effects on osteoblast differentiation are seen with the BET inhibitor +JQ1, and this effect is reversible upon its removal indicating that this small molecule has no lasting effects on the differentiation capacity of MC3T3 cells. Mechanistically, we find that Brd4 binds at known Runx2 binding sites in promoters of bone-related genes. Collectively, these findings suggest that Brd4 is recruited to osteoblast-specific genes and may cooperate with bone-related transcription factors to promote osteoblast lineage commitment and maturation.

The effect of adipose-derived stem cells on enthesis healing after repair of acute and chronic massive rotator cuff tears in rats.

BACKGROUND: Chronic massive rotator cuff tears heal poorly and often retear. This study investigated the effect of adipose-derived stem cells (ADSCs) and transforming growth factor-ß3 (TGF-ß3) delivered in 1 of 2 hydrogels (fibrin or gelatin methacrylate [GelMA]) on enthesis healing after repair of acute or chronic massive rotator cuff tears in rats. METHODS: Adult male Lewis rats underwent bilateral transection of the supraspinatus and infraspinatus tendons with intramuscular injection of botulinum toxin A (n = 48 rats). After 8 weeks, animals received 1 of 8 interventions (n = 12 shoulders/group): (1) no repair, (2) repair only, or repair augmented with (3) fibrin, (4) GelMA, (5) fibrin + ADSCs, (6) GelMA + ADSCs, (7) fibrin + ADSCs + TGF-ß3, or (8) GelMA + ADSCs + TGF-ß3. An equal number of animals underwent acute tendon transection and immediate application of 1 of 8 interventions. Enthesis healing was evaluated 4 weeks after the repair by microcomputed tomography, histology, and mechanical testing. RESULTS: Increased bone loss and reduced structural properties were seen in chronic compared with acute tears. Bone mineral density of the proximal humerus was higher in repairs of chronic tears augmented with fibrin + ADSCs and GelMA + ADSCs than in unrepaired chronic tears. Similar improvement was not seen in acute tears. No intervention enhanced histologic appearance or structural properties in acute or chronic tears. CONCLUSIONS: Surgical repair augmented with ADSCs may provide more benefit in chronic tears compared with acute tears, although there was no added benefit to supplementing ADSCs with TGF-ß3.

Predictive validity of the HCR-20 for inpatient aggression: the effect of intellectual disability on accuracy.

BACKGROUND: People with intellectual disability (ID) account for a large proportion of aggressive incidents in secure and forensic psychiatric services. Although the Historical, Clinical, Risk Management 20 (HCR-20) has good predictive validity in inpatient settings, it does not perform equally in all groups and there is little evidence for its efficacy in those with ID. METHOD: A pseudo-prospective cohort study of the predictive efficacy of the HCR-20 for those with ID (n = 109) was conducted in a UK secure mental health setting using routinely collected risk data. Performance of the HCR-20 in the ID group was compared with a comparison group of adult inpatients without an ID (n = 504). Analysis controlled for potential covariates including security level, length of stay, gender and diagnosis. RESULTS: The HCR-20 total score was a significant predictor of any aggression and of physical aggression for both groups, although the area under the curve values did not reach the threshold for a large effect size. The clinical subscale performed significantly better in those without an ID compared with those with. The ID group had a greater number of relevant historical and risk management items. The clinicians' summary judgment significantly predicted both types of aggressive outcomes in the ID group, but did not predict either in those without an ID. CONCLUSIONS: This study demonstrates that, after controlling for a range of potential covariates, the HCR-20 is a significant predictor of inpatient aggression in people with an ID and performs as well as for a comparison group of mentally disordered individuals without ID. The potency of HCR-20 subscales and items varied between the ID and comparison groups suggesting important target areas for improved prediction and risk management interventions in those with ID.

Digestibility and metabolic utilization of diets containing whole-ear corn silage and their effects on growth and slaughter traits of heavy pigs.

The aim was to evaluate 2 levels of dietary inclusion of chopped whole-ear corn silage (WECS) on energy and nutrient utilization, growth, and slaughter performances of heavy pigs. Two in vivo experiments were conducted to determine digestibility and metabolic utilization of WECS using 18 barrows weighing 118 ± 8 kg BW on average, metabolic cages and respiration chambers (Exp. 1), and the effect of WECS on the growth performance and carcass traits on 42 barrows from 90 to 170 kg BW (Exp. 2). In both experiments, pigs were fed 3 experimental diets: a control diet (CON) containing cereal meals, extracted soybean meal, and wheat bran (80%, 9%, and 8% of DM, respectively) and 2 diets containing 15% (15WECS) or 30% WECS (30WECS) on a DM basis in place of wheat bran and corn meal. The diets were prepared daily by mixing the WECS to a suitable compound feed. Feed intake was always restricted to allow a daily DMI of 7.2% BW(0.75) in Exp. 1 and from 8.0% to 6.5% BW(0.75) in Exp. 2. Diets had similar NDF contents (15.2% to 15.8% of DM), and WECS inclusion resulted in a slight reduction in CP content (from 14.0% to 13.6% of DM) and a considerable decrease in P content (from 0.47% to 0.30% of DM). Digestibility of OM, CP, and fat was similar among diets, whereas P digestibility was lower (P < 0.05) for the 30WECS diet (33.5%) in comparison with the CON and 15WECS diets (45.5% and 44.1%, respectively). Nitrogen lost in feces and urine and N retained were not different among diets, whereas P retained decreased with the increase of WECS (5.4, 3.7, and 2.2 g/d for the CON, 15WECS, and 30WECS diets, respectively; P < 0.05). No difference among diets was observed for energy balance. The WECS contained 13.48 MJ ME and 9.39 MJ NE/kg DM. In Exp. 2, feed intake was not depressed by WECS inclusion, and the ADG for the whole experiment was not different among dietary treatments (from 737 to 774 g/d). Fecal pH was lower (P < 0.05) for the WECS diets than the control diet (7.10 and 7.00 vs. 7.40) and for the sampling at 150 kg BW than that at 130 and 110 kg BW (6.96 vs. 7.29 and 7.24). At slaughter, lean percentage in the carcass was lower in the 30WECS diet than those of the other 2 diets (46.8% vs. 48.3% and 48.6%, P = 0.05). The overall experimental data obtained in both trials indicate that substitution of wheat bran and corn meal for WECS (up to 30% of DM) does not affect, with the exception of P utilization and carcass leanness, energy and nutrient utilization and performance of heavy pigs in the last phase of growing.

Ultrasonographic appearance of the coelomic cavity in healthy green iguanas.

OBJECTIVE: To describe the ultrasonographic appearance of the coelomic cavity in healthy green iguanas. DESIGN: Cross-sectional study. ANIMALS: 26 healthy green iguanas (20 males and 6 females). PROCEDURES: For coelomic ultrasonography, animals were physically restrained in dorsal recumbency by an assistant; chemical restraint was not used. Qualitative and quantitative observations were recorded. RESULTS: Structures that could be visualized in all animals included the heart and cardiac chambers; liver; caudal vena cava; hepatic veins; portal vein; gallbladder; pyloric portion of the stomach; and, when distended, urinary bladder. Visualization of the kidneys was poor. The spleen could be identified in 17 animals, and the gonads could be identified in 22, but were most easily identified in males evaluated during November (ie, during the breeding season); no females were evaluated during the breeding season. Physiologic enlargement of the testes yielded an acoustic window for the spleen by displacing overlying intestine. Anechoic, free coelomic fluid was identified in 3 animals. Measurements of overall cardiac size, ventricular wall thickness, gallbladder size, thickness of the pyloric portion of the stomach, and splenic size were obtained. Only ventricular wall thickness was significantly correlated with body weight. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that ultrasonography allowed examination of most coelomic structures in green iguanas. The procedure was easily performed and was well tolerated in conscious animals.

From policy to practice: challenges in infant feeding in emergencies during the Balkan crisis.

The preparation and dissemination of policy statements are necessary but insufficient to prevent the inappropriate use of infant-feeding products in emergencies. The widespread failure of humanitarian agencies operating in the Balkan crisis to act in accordance with international policies and recommendations provides a recent example of the failure to translate infant-feeding policies into practice. This article explores the underlying reasons behind the failures which include: (1) the weak institutionalisation of policies; (2) the massive quantities of unsolicited donations of infant-feeding products: (3) the absence of monitoring systems; (4) inadequate co-ordination mechanisms; (5) the high costs of correcting mistakes; and (6) the cumulative effects of poor practice. Efforts to uphold best practice during the crisis are also documented. Finally, the article identifies actions that could be undertaken in advance of and during future emergencies to enhance the application of infant feeding policies in emergencies.

The management of severe malnutrition in Burundi: an NGO's perspective of the practical constraints to effective emergency and medium-term programmes.

The NGO, Action contre la Faim (ACF), has been operating in Burundi since 1994, where the situation has dramatically hampered humanitarian programmes. These include poor security resulting in an inability to maintain 24-hour care for the severely malnourished and poor access to the beneficiaries, all within a politically and economically unstable context. However, ACF has been able to capitalise on lessons learnt and reflect on ways to move forward, which have included the improvement of their capacities in the treatment of severe malnutrition and the integration within broader disciplines and national structures. The protocols for the treatment of severe malnutrition currently used in Burundi have been developed as a result of the research of ACF over the past five years. Other aspects of the nutrition programme remain to be further developed. These include, for example, an improved capacity to care for severely malnourished adolescents and adults; and also a better knowledge of the beneficiaries to allow for stronger links between the provision of treatment and support for their longer-term food security.

A basic list of recommended books and journals for support of clinical dentistry in a nondental library.

A basic list of 133 book and journal titles in dentistry is presented. The list is intended as a bibliographic selection tool for those libraries and health institutions that support clinical dentistry programs and services in the nondental school environment in the United States and Canada. The book and journal titles were selected by the membership of the Dental Section of the Medical Library Association (MLA). The Dental Section membership represents dental and other health sciences libraries and dental research institutions from the United States and Canada, as well as from other countries. The list was compiled and edited by the Ad Hoc Publications Committee of the Dental Section of MLA. The final list was reviewed and subsequently was approved for publication and distribution by the Dental Section of MLA during the section's 1996 annual meeting in Kansas City, Missouri.

The production of alamethicins by Trichoderma spp.

The production of polypeptides containing a high percentage of 2-methylalanine residues by a number of isolates of Trichoderma spp. has been examined. It has been shown that good yields (0.5-1.0 g L-1) can be achieved on synthetic media provided an insoluble carbohydrate is included and provided single-spore isolates that have this production ability are selected from time to time. Such yields could not be obtained on any single nitrogen source investigated, but a mixture of potassium nitrate, glutamine, and 2-methylalanine was effective. It was shown that at least eight polypeptides were produced in shake-flask or tank fermentation and that the proportions of these metabolites depended on the fermentation temperature, its pH, age, and aeration. Fermentation conditions for enhancing the production (independently) of two of the metabolites at the expense of the others are given. These two metabolites have been obtained in crystalline form and details of some of their physical and chemical properties are given.

Caffeine-induced taste aversion and mimetic responses.

Novel tastes preceded a range of caffeine doses (10-80 mg/kg) in a taste aversion training trial. One week later rats which had doses of 30 mg or higher showed strong aversions as measured by a single bottle consumption test. The 10 and 20 mg dose produced the most hyperactivity and apparently enhanced intake of the taste paired with caffeine. During the training trial, rats receiving the 80 mg dose exhibited copious gapes and chin-rubs, mimetic responses to noxious tastes. Gapes also occurred in these subjects during the aversion test. Consumption was more sensitive than mimetic responding as a measure of the aversive effects of caffeine. Only the 80 mg dose produced neophobia. Tests with isotonic injections indicated that tonicity was not the source of the aversions.