Comment on Martínez-Delgado et al. Using Absorption Models for Insulin and Carbohydrates and Deep Leaning to Improve Glucose Level Predictions. Sensors 2021, 21, 5273.

The paper "Using Absorption Models for Insulin and Carbohydrates and Deep Leaning to Improve Glucose Level Predictions" (Sensors2021, 21, 5273) proposes a novel approach to predicting blood glucose levels for people with type 1 diabetes mellitus (T1DM). By building exponential models from raw carbohydrate and insulin data to simulate the absorption in the body, the authors reported a reduction in their model's root-mean-square error (RMSE) from 15.5 mg/dL (raw) to 9.2 mg/dL (exponential) when predicting blood glucose levels one hour into the future. In this comment, we demonstrate that the experimental techniques used in that paper are flawed, which invalidates its results and conclusions. Specifically, after reviewing the authors' code, we found that the model validation scheme was malformed, namely, the training and test data from the same time intervals were mixed. This means that the reported RMSE numbers in the referenced paper did not accurately measure the predictive capabilities of the approaches that were presented. We repaired the measurement technique by appropriately isolating the training and test data, and we discovered that their models actually performed dramatically worse than was reported in the paper. In fact, the models presented in the that paper do not appear to perform any better than a naive model that predicts future glucose levels to be the same as the current ones.

Paced Mating Behaviour Is Influenced by Duration of Female Post-Ejaculatory Interval.

BACKGROUND: Laboratory paradigms that enable the female rat to control the frequency and temporal distribution of sexual stimulation are well-suited to gaining knowledge about female sexual function; however, the variety of procedures used influence the specific behaviors exhibited by female rats and bring uncertainty into the conclusions that can be drawn. AIM: In this study, we evaluated the effects of test parameters on the display of paced mating behavior in female rats to develop better preclinical models for exploring female sexual health. METHODS: Sexually experienced, estradiol- and progesterone-primed female rats were tested under paced mating conditions to determine whether sexual behavior differed as a function of number of male partners (1 or 3; Experiment 1), the time span between receipt of an ejaculation and the next intromission (ie, the post-ejaculatory interval or PEI; Experiment 2), or the duration of ejaculations (Experiment 3). OUTCOMES: Contact-return latency, exit latency, and withdrawal duration after mounts, intromissions, and ejaculations. RESULTS: The shorter withdrawal latency after intromission and longer return latency after ejaculation observed in 30-minute paced mating tests is best attributed to the female's PEI. The duration of the PEI is a specific factor that affects the display of paced mating behavior. CLINICAL TRANSLATION: Understanding of neurobiological mechanisms and sensory factors influencing sexual behavior learned from these models can be applied to studies of human female sexual health. STRENGTHS & LIMITATIONS: Sexual motivation is indicated by analogous behaviors and supported by evolutionarily conserved systems in women and rats, meaning that animal models can be used to identify neurobiological mechanisms subserving these behaviors. Nonetheless, data related specifically to physiological effects of different mating paradigms are needed in future research. CONCLUSION: Procedures that mimic naturalistic settings, and thus enable female sexual motivation to drive behavior, are apt for studies aimed at understanding mechanisms supporting female sexual function whereas artificially lengthening the interval between an ejaculation and subsequent intromission may provide a model to study female sexual dysfunction. Corlett AG, Frankl PR, Akindona FAB, et al. Paced Mating Behaviour Is Influenced by Duration of Female Post-Ejaculatory Interval. J Sex Med 2022;19:1506-1516.

Sexual experience confers resilience to restraint stress in female rats.

During paced mating, sexually experienced female rats spend more time with the male, return to the male more quickly after intromission, and exhibit shorter interintromission intervals as compared to sexually naïve rats. Factors that trigger the shift in paced mating behavior are unknown. The present study used the elevated plus maze to test whether anxiety-like behavior differs as a function of sexual experience. Ovariectomized, Long-Evans female rats were primed with estradiol benzoate plus progesterone (EB + P) and then either received four, twice weekly, paced mating treatments to gain sexual experience (Experienced) or remained sexually naïve (Naïve) but were exposed to an empty mating apparatus. In Experiment 1, anxiety-like behavior was compared between Experienced or Naïve female rats that were primed with either EB + P or oil. Significantly more time was spent in open arms under EB + P vs. oil, independent of sexual history. To test whether exposure to an acute stressor before elevated plus maze testing affected anxiety-like behavior, EB + P treated, Experienced or Naïve rats received paced mating (Experiment 2) or restraint (Experiment 3) immediately prior to the elevated plus maze task. Restraint, but not mating, led to less anxiety-like behaviors for Experienced rats compared to Naïve rats. Collectively, our data shows that one component of the shift in paced mating behavior that occurs with sexual experience appears to be altered stress responsiveness. We propose that mating is a beneficial stressor that, when repeated, increases the ability to cope with anxiety-producing events such as aversive components of mating or non-voluntary stressors.

Previous sexual experience alters the display of paced mating behavior in female rats.

The present study tested whether the display of paced mating behavior in female rats over four weekly tests is affected by sexual experience and whether test parameters, i.e., ending the test based on time or number of stimulations received, influence behavioral changes. In Experiment 1A rats with nonpaced sexual experience returned to the male more quickly overall compared to sexually naïve rats in a 30-min test of paced mating behavior. In Experiment 1B, rats received four weekly 30-min tests with one, different, male rat partner each week. Over the four tests, rats returned to the male significantly more quickly after intromissions, but significantly more slowly after ejaculations. Experiment 2A tested whether sexual experience would influence paced mating behavior in tests with a 15-intromission end criterion and the male replaced after ejaculation. Rats tested weekly under 15-intromission test conditions returned to the male significantly more quickly after intromissions, but no behavioral change was observed after ejaculations. When those same rats were given a 30-min test of paced mating behavior (Experiment 2B), they returned to the male significantly more slowly after ejaculations. Collectively, these data show that sexual experience influences the display of paced mating behavior in female rats and that the test parameters interact with sexual experience to influence the nature of the changes. Sexual experience may facilitate behaviors that promote reproductive success in female rats.

Male Syrian hamsters demonstrate a conditioned place preference for sexual behavior and female chemosensory stimuli.

Sexual behavior is a natural reward for many rodent species, and it often includes chemosensory-directed components. Chemosensory stimuli themselves may also be rewarding. Conditioned place preference (CPP) is one paradigm frequently used to test the rewarding properties of a range of stimuli. Males and females of several rodent species show a CPP for sexual behavior; however, it is currently unknown whether sexual behavior can induce a CPP in male Syrian hamsters. As male Syrian hamsters are an animal model commonly used for investigation of the neurobiology of sexual behavior, understanding the rewarding components of sexual stimuli will better direct future research on brain regions and neurotransmitters involved in these behaviors. Experiment 1 tested the prediction that male hamsters show a CPP for sexual behavior. Female chemosensory stimuli are essential for the display of sexual behavior in male hamsters; however, the rewarding properties of female chemosensory stimuli contained in vaginal secretions (VS) are uncertain. Therefore, experiment 2 tested the prediction that male hamsters show a CPP for VS. This study is the first demonstration that both sexual behavior and VS induce a CPP in male hamsters. Thus, female chemosensory stimuli are a natural reward in a species that is dependent on these stimuli for reproductive fitness.

II. Antidepressants and sexual behavior: Acute fluoxetine, but not ketamine, disrupts paced mating behavior in sexually experienced female rats.

Female sexual dysfunction is both a symptom of depression and exacerbated by treatments for depression. Ketamine, a novel treatment for depression, has been shown to enhance, whereas fluoxetine has been shown to impair sexual motivation. Sexual experience leads to more robust partner preference and paced mating behavior in female rats. Whether acute ketamine and fluoxetine similarly affect sexual motivation and mating behavior in sexually experienced female rats is unknown. Sexually experienced female rats received 10 mg/kg i.p. of ketamine or saline vehicle (Experiment 1) or 10 mg/kg i.p. of fluoxetine or water vehicle (Experiment 2) 30 min before a 10-min no-contact partner preference test followed immediately by a 15-intromission paced mating test. Partner preference and paced mating behavior did not differ between ketamine- and saline-treated rats. In contrast, rats treated with fluoxetine spent significantly less time with either stimulus animal and were less active during the partner preference test than water-treated rats. Additionally, contact-return latency to ejaculation was significantly longer in fluoxetine-treated rats and they spent less time with the male during paced mating in comparison to water-treated rats. Thus, even with sexual experience, fluoxetine disrupts sexual function whereas ketamine has no detrimental effects on sexual behavior in female rats. A growing body of evidence suggests that ketamine is an encouraging new approach to treat depression particularly because it is not associated with sexual dysfunction.

Sexual Behavior is Enhanced by Regular, Repeated Mating from Young Adulthood to Middle Age in Female Long-Evans Rats.

BACKGROUND: Aging is associated neuroendocrine changes in women. Animals can be used to model these changes, as well as changes in reproductive behavior. OBJECTIVE: The current study was designed to characterize mating behavior across age and assess the effects of age and sexual history on mating behavior. METHODS: Sexual motivation was assessed using the partner-preference test, in which a female rat is given the choice to interact with a same-sex conspecific or a sexually-vigorous male rat, with which she can mate. RESULTS: Across repeated mating tests (2-12 months of age), female rats spent more time with the male, displayed more solicitation behaviors, were less likely to leave the male after mounts, but visited both stimulus animals less frequently. Comparing a separate group of age-matched, hormoneyoked female rats mated for the first time at 12 months of age to female rats mated for the first time at 2 months of age showed that the 12 month rats visited both stimulus animals less, were less likely to leave the male after mounts, took longer to return to the male after mounts, and displayed fewer solicitation behaviors than their younger counterparts. Relative to middle-aged female rats once they were sexually experienced, 12 month naïve rats spent less time with the male, were more likely to leave the male after mounts, and displayed fewer solicitation behaviors. Furthermore, 12 month naïve rats failed to discriminate between the stimulus animals, visiting both stimulus animals at the same rate unlike 2 month naïve or 12 month experienced rats. CONCLUSION: Taken together, these results suggest that aging affects some measures of sexual behavior, but most effects of age can be mitigated by regular, repeated mating.

I. Antidepressants and sexual behavior: Weekly ketamine injections increase sexual behavior initially in female and male rats.

The present study characterized the effects of weekly ketamine injections on sexual behavior and anxiety in female and male rats, using a dosing protocol that mimics human therapeutic treatment for depression. In Experiment 1A, ketamine (10 mg/kg, i.p.) or saline was injected once per week for four consecutive weeks. The partner preference paradigm was used to measure sexual motivation 30 min after each weekly injection. Briefly, subjects were first given a 10-min test during which they could choose to spend time in the vicinity of a sexually receptive female stimulus or a sexually experienced male stimulus, however physical contact was restricted (no-contact). Immediately after, subjects were given unrestricted access to the stimulus animals (contact). After a washout period, subjects received four additional weekly injections of ketamine or saline, and then were tested for anxiety-like behavior on the elevated plus maze (EPM) after the last injection (Experiment 1B). For Experiment 2, similar procedures were used to test the effects of weekly ketamine injections on sexual motivation (Experiment 2A) and anxiety (Experiment 2B) in male subjects. In female subjects, ketamine increased sexual motivation as measured by greater time spent with the male stimulus, decreased likelihood of leaving after receiving mounts, and shorter return latencies after receiving intromissions, when compared to saline controls. In male subjects, ketamine shortened latency to first mount and first intromission, as well as increased time spent with the female stimulus. Very little anxiety was observed in either group (ketamine or saline) of female or male subjects when tested on the EPM. In conclusion, even after four weeks of ketamine exposure, sexual dysfunction did not emerge in either females or males. In contrast, ketamine increased sexual motivation in both females and males, with an initial robust response. However, as both groups gained sexual experience, the impact of ketamine diminished.

Conditioned place preference for mating is preserved in rats with pelvic nerve transection.

Female rats exhibit a conditioned place preference (CPP) for a context paired with mating. The present experiment tested the hypothesis that the activation of the pelvic nerve mediates the reinforcing effects of mating for female rats. Rats underwent bilateral pelvic nerve or sham transection and then received paced mating, nonpaced mating, or the control treatment during a CPP procedure. Pelvic nerve transection did not affect the CPP for paced or nonpaced mating. In tests of paced mating behavior, contact-return latencies following intromissions were significantly shorter in rats with pelvic nerve transection than they were in rats with sham transections. These results show that the pathway conveying the reinforcing effects of mating stimulation does not depend on the integrity of the pelvic nerve, but that activation of the pelvic nerve contributes to the display of paced mating behavior.

Artificial vaginocervical stimulation induces a conditioned place preference in female rats.

Female rats express a conditioned place preference (CPP) for a context paired with mating. During a mating encounter, the female rat is exposed to several different types of stimuli, including, but not limited to, vaginocervical stimulation and social contact. The present experiment tested the hypothesis that two components of the mating interaction, vaginocervical stimulation or social contact, each induce a CPP in female rats. During conditioning rats received nonpaced mating, artificial vaginocervical stimulation, social interaction or a control treatment. Rats expressed a CPP for the context paired with nonpaced mating or artificial vaginocervical stimulation whereas social interaction and the control treatment did not induce a CPP. The present findings highlight the important role that vaginocervical stimulation plays in the reinforcing effects of mating in female rats.

Zaprinast, a phosphodiesterase type-5 inhibitor, alters paced mating behavior in female rats.

Nitric oxide (NO) is the primary mediator of blood flow in female genital tissues and drugs that enhance the activity of nitric oxide, such as phosphodiesterase type-5 (PDE-5) inhibitors, increase vaginal blood flow in anesthetized rats. The goal of the present study was to test the effects of one PDE-5 inhibitor, zaprinast, on the display of sexual behaviors in gonadectomized, estrogen- and progesterone-treated female rats. Experiment 1 demonstrates that zaprinast alters paced mating behavior by lengthening the contact-return latency to ejaculation; there is a significant relationship between dose of zaprinast (range 1.5-6 mg/kg) and contact-return latency to ejaculation. Experiment 2 illustrates that zaprinast has no effect on preference for an intact male as measured in a No Contact partner preference test. Rats receiving zaprinast tend to exhibit reduced locomotor activity in both experiments. Collectively, these findings demonstrate that modulation of the NO-cGMP pathway using a PDE-5 inhibitor alters the display of paced mating behaviors in rats.

Sexually experienced, but not naïve, female rats show a conditioned object preference (COP) for mating after a single training trial.

Female rats with mating experience spend more time with the male rat, exhibit shorter contact-return latency to intromission, and display more proceptive behaviors in the male rat's compartment than during the first mating experience. The present study tested 1) whether mating induced conditioned object preference (COP) is possible with a single conditioning trial and 2) whether a preference is induced for an object associated with the first mating encounter or the fifth mating encounter in female rats. Ovariectomized, Long-Evans female rats were primed with estradiol benzoate + progesterone and either exposed to an empty paced mating chamber for 15 min (Naïve) or received a 15 intromission test of paced mating behavior (Experienced) on four separate occasions before undergoing the COP procedure. Experienced, but not Naïve, female rats developed a COP for a single mating bout, indicating that mating is highly rewarding for sexually experienced female rats. The findings raise questions about the effect of sexual experience on reward regions in the brain, the responsiveness of genital tissue, and learning mechanisms.

Prepubertal ovariectomy modulates paced mating behavior but not sexual preference or conditioned place preference for mating in female rats.

The present study investigated whether the presence or absence of peripubertal ovarian hormones affects sexual preference and conditioned place preference for paced mating in adult female rats primed with 10µg estradiol benzoate and 1mg progesterone. Ovariectomy (OVX) occurred either before or after pubertal development, and 4weeks later rats began a series of behavioral tests. Rats with ovaries removed before the pubertal timeframe (Prepubertal OVX) were more active, more likely to withdrawal from the male compartment, and did not discriminate between mounts and intromissions during paced mating relative to rats with ovaries during puberty (Adult OVX). Both Adult OVX and Prepubertal OVX rats showed a higher preference for the male when hormone primed vs. oil treated and a conditioned place preference for paced mating behavior. The results of the present study demonstrate that some, but not all, aspects of female sexual behavior require ovarian hormones during puberty.

Sex- and age-specific effects of anabolic androgenic steroids on reproductive behaviors and on GABAergic transmission in neuroendocrine control regions.

Illicit use of anabolic androgenic steroids (AAS) has become a prevalent health concern not only among male professional athletes, but, disturbingly, among a growing number of women and adolescent girls. Despite the increasing use of AAS among women and adolescents, few studies have focused on the effects of these steroids in females, and female adolescent subjects are particularly underrepresented. Among the hallmarks of AAS abuse are changes in reproductive behaviors. Here, we discuss work from our laboratories on the actions of AAS on the onset of puberty and sexual behaviors in female rodents, AAS interactions and sex- and age-specific effects of these steroids on neural transmission mediated by gamma-aminobutyric acid receptors within forebrain neuroendocrine control regions that may underlie AAS-induced changes in these behaviors.

Stimulus animal characteristics do not modulate the expression of partner preference by female rats.

The stimulus animals used in tests of partner preference in female rats vary. To test the hypothesis that the alternative stimulus animal modulates the preference for an intact male, gonadectomized (GDX) female rats received estradiol benzoate plus progesterone or the oil vehicle and were tested for partner preference with either an intact male paired with a GDX hormone-primed female or an intact male paired with a GDX male. Rats were tested under conditions that limited physical contact (No Contact) or allowed sexual interaction (Contact). Stimulus animal condition was not a primary determinant of partner preference. In contrast, contact condition and hormone treatment modulated preference, as well as activity levels and the display of proceptive behaviors. Our findings demonstrate that the characteristics of the alternative stimulus animals tested here do not play a significant role in modulating partner preference in female rats.

Sexual experience modulates partner preference and mPOA nitric oxide synthase in female rats.

Sexually experienced female rats return to the male more quickly after intromissions, exhibit shorter interintromission intervals, and spend more time with the male rat during a test of paced mating behavior in comparison to naïve rats. The present study tested whether these changes reflect heightened sexual motivation independent of receipt of vaginocervical stimulation and/or neurochemical changes in the medial preoptic area (mPOA). Ovariectomized, female rats were given estradiol benzoate and progesterone, and then received either 6 paced mating encounters (experienced) or 6 control exposures to an empty paced mating arena (naïve). Experienced and naïve rats received a no-contact partner preference test under oil vehicle and then under hormone on a different day. Hormonal status and sexual experience led to significantly higher preference for the male. Brains were collected 1 hr after both experienced and naïve rats received paced mating to compare mPOA levels of Fos, a marker of neural activity, in response to copulation and nitric oxide synthase (NOS), the enzyme responsible for production of nitric oxide (NO). Expression of NOS was higher in experienced relative to naïve rats, whereas Fos was comparable between the groups. The data are consistent with the idea that both sexual motivation and changes to the mPOA contribute to the shift in paced mating behavior induced by sexual experience. (PsycINFO Database Record

Paced mating behavior is affected by clitoral-vaginocervical lidocaine application in combination with sexual experience.

The present study tested the effects of lidocaine anesthetic ointment applied to the vaginocervical (Experiment 1) or clitoral-vaginocervical (Experiment 2) areas on the display of paced mating behavior over the course of five weekly tests in ovariectomized, hormone-primed, Long-Evans rats. Experiment 3 tested whether rats that acquired sexual experience without ointment application would exhibit altered paced mating behavior on a fifth test under clitoral-vaginocervical lidocaine or vehicle application. Although rats in Experiment 1 and Experiment 2 exhibited shorter contact-return latencies after intromission and reduced likelihood of leaving the male compartment following mounts and intromissions after gaining sexual experience, only rats that received clitoral-vaginocervical lidocaine exhibited altered paced mating behavior relative to vehicle. Specifically, clitoral-vaginocervical lidocaine resulted in shorter contact-return latency to ejaculation and greater percentage of time with the male. Paced mating behavior of sexually experienced rats in Experiment 3 was not disrupted when tested after clitoral-vaginocervical lidocaine treatment. Together, these studies suggest that the sensory input during repeated mating encounters affects the pattern of paced mating behavior that develops with sexual experience.

Female rats express a conditioned object preference for receipt of sexual stimulation.

Female rats alternately approach and avoid the male rat during copulation, potentially reflecting appetitive and aversive aspects of mating, respectively. We developed a novel classical conditioning procedure, conditioned object preference (COP), to test whether female rats show increased approach toward a conditioned stimulus associated directly with receipt of sexual stimulation. During conditioning, one scented object was paired with an appetitive stimulus and a different object plus scent was paired with a control stimulus on a separate day. After conditioning, preference for each object was evaluated with a choice task. Experiment 1 was conducted to verify the procedure. Rats exhibited a significant COP for 1mg/kg amphetamine, indicating that the conditioned object preference procedure is an effective tool for evaluating the rewarding nature of a treatment. In Experiment 2, paced mating to one ejaculation and experimenter-delivered artificial vaginocervical stimulation (aVCS) each induced a COP. The robust COPs for paced mating and aVCS support the notion that female rats experience a reward state during receipt of sexual stimulation. Moreover, the data suggest that any aversive aspects of receipt of sexual stimulation do not overshadow the appetitive effects.

Adolescent brain maturation is necessary for adult-typical mesocorticolimbic responses to a rewarding social cue.

The interpretation of social cues must change during adolescence in order to promote appropriate social interactions in adulthood. For example, adult, but not juvenile, male Syrian hamsters find female pheromones contained in vaginal sections (VS) rewarding, and only adult hamsters engage in sexual behavior with a receptive female. We previously demonstrated that the rewarding value of VS is both testosterone- and dopamine-dependent. Additionally, VS induces Fos expression throughout the mesocorticolimbic circuit in adult but not juvenile hamsters. In this study, we determined whether or not treatment of juvenile male hamsters with testosterone is sufficient to promote adult-like neural responses to VS. Juvenile and adult male hamsters were gonadectomized and given empty or testosterone-filled subcutaneous capsules for 1 week. Hamsters were then exposed to either clean or VS-containing mineral oil on their nares, and brains were collected 1 h later for immunohistochemistry to visualize Fos and tyrosine hydroxylase immunoreactive cells. Testosterone treatment failed to promote adult-typical patterns of Fos expression in juvenile hamsters; indeed, in some brain regions, juveniles exposed to VS expressed less Fos compared to age-matched controls while, as expected, adults exposed to VS expressed greater Fos compared to age-matched controls. Age-related changes in tyrosine hydroxylase expression were also observed. These data indicate that testosterone cannot activate the adult-typical pattern of Fos expression in response to female social cues in prepubertal males, and that additional neural maturation during adolescence is required for adult-typical mesocorticolimbic responses to female pheromones.

An intact medial preoptic area is necessary for zaprinast to modulate paced mating behavior in female rats.

The present study examined the interaction between the regulation of paced mating behavior by the medial preoptic area (mPOA) and by the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) pathway, as modulated by zaprinast, a phosphodiesterase inhibitor. Rats receiving mPOA or sham lesions were tested for paced mating behavior. Subsequently, rats were treated with zaprinast (3 mg/kg) before a second paced mating test. The expected lengthening of contact-return latencies following intromissions and ejaculations was observed in rats with mPOA lesions relative to rats with sham lesions. In addition, rats with sham lesions responded to zaprinast with a lengthening of contact-return latency following ejaculation. Contact-return latencies did not change in response to zaprinast in rats with mPOA lesions. These results demonstrate that the alterations in paced mating behavior observed in rats with mPOA lesions persist despite manipulation of the nitric oxide-cyclic guanosine monophospate pathway.