Evidence Of Sexual Dimorphism Research Articles

TVNS alters inflammatory response in adult VPA-induced mouse model of autism: evidence for sexual dimorphism.

Autism is a neurodevelopmental disorder with limited treatment alternatives and which incidence is increasing. Some research suggests that vagus nerve simulation might lead to the reduction of certain symptom. Therefore, we aimed to examine the effect of bilateral transcutaneous auricular vagus nerve stimulation (tVNS) on the inflammatory response in an adult valproic acid (VPA) induced mouse (C57BL6) model of autism for the first time. The autism model was induced by oral VPA administration (600 mg·kg-1) to C57BL/6 pregnant mice on E12.5 days. The study included three groups: the VPA Transcutaneous Auricular Stimulation Group (VPA + tVNS), the VPA Control Group (VPA + sham), and the Healthy Control Group (Control + sham). Each group included 16 mice (8 M/8 F). Our results show that serum IL-1β and IL-6 levels were significantly higher in male VPA-exposed mice than controls. However, IL-1β was significantly lower, and IL-6, TNF- α, and IL-22 were not different in female VPA-exposed mice compared to the control group. Brain NLRP3 levels were significantly higher in both sexes in the VPA autism model (P < 0.05). tVNS application increased brain NLRP3 levels in both sexes and reduced serum IL-1β levels in male mice. We conclude that cytokine dysregulation is associated with the VPA-induced adult autism model, and the inflammatory response is more pronounced in male mice. tVNS application altered the inflammatory response and increased brain NLPR3 levels in both sexes. Further studies are needed to understand the beneficial or detrimental role of the inflammatory response in autism and its sexual dimorphism.

Alterations in placental mitophagy with maternal obesity and gestational diabetes: evidence for sexual dimorphism

Alterations in placental mitophagy with maternal obesity and gestational diabetes: evidence for sexual dimorphism

Genetic Architecture and Analysis Practices of Circulating Metabolites in the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program.

Circulating metabolite levels partly reflect the state of human health and diseases, and can be impacted by genetic determinants. Hundreds of loci associated with circulating metabolites have been identified; however, most findings focus on predominantly European ancestry or single study analyses. Leveraging the rich metabolomics resources generated by the NHLBI Trans-Omics for Precision Medicine (TOPMed) Program, we harmonized and accessibly cataloged 1,729 circulating metabolites among 25,058 ancestrally-diverse samples. We provided recommendations for outlier and imputation handling to process metabolite data, as well as a general analytical framework. We further performed a pooled analysis following our practical recommendations and discovered 1,778 independent loci associated with 667 metabolites. Among 108 novel locus - metabolite pairs, we detected not only novel loci within previously implicated metabolite associated genes, but also novel genes (such as GAB3 and VSIG4 located in the X chromosome) that have putative roles in metabolic regulation. In the sex-stratified analysis, we revealed 85 independent locus-metabolite pairs with evidence of sexual dimorphism, including well-known metabolic genes such as FADS2 , D2HGDH , SUGP1 , UTG2B17 , strongly supporting the importance of exploring sex difference in the human metabolome. Taken together, our study depicted the genetic contribution to circulating metabolite levels, providing additional insight into the understanding of human health.

Changes in maternal blood and placental lipidomic profile in obesity and gestational diabetes: Evidence for sexual dimorphism.

Obesity and gestational diabetes (GDM) are associated with adverse pregnancy outcomes and program the offspring for cardiometabolic disease in a sexually dimorphic manner. The placenta transfers lipids to the fetus and uses these substrates to support its own metabolism impacting the amount of substrate available to the growing fetus. We collected maternal plasma and placental villous tissue following elective cesarean section at term from women who were lean (pre-pregnancy BMI 18.5-24.9), obese (BMI>30) and type A2 GDM (matched to obese BMI) with male or female fetus (n=4 each group). Lipids were extracted and fatty acid composition of different lipid classes were analyzed by LC-MS/MS analysis. Significant changes in GDM vs obese, GDM vs lean, and obese vs lean were determined using t-test with a Tukey correction set at p<0.05. In placental samples 436 lipids were identified, among which 85 showed significant changes. Of note only in male placentas significant decreases in C22:6 - docosahexaenoic acid (DHA) in phosphatidylcholine (PC) and triglyceride lipid species were seen when comparing tissue from GDM women to lean. In maternal plasma we observed no effect of obesity. GDM or fetal sex. This is the first study assessing fatty acid composition of lipids in matched maternal plasma and placental tissue from lean, obese, and GDM women stratified by fetal sex. It highlights how GDM affects distribution of fatty acids in lipid classes changes in a sexually dimorphic manner in the placenta.

Sexual selection in seaweed? Testing Bateman's principles in the red alga Gracilaria gracilis.

In anisogamous species, sexual selection is expected to be stronger in males. Bateman's principles state that the variance in (i) reproductive and (ii) mating success is greater for males, and (iii) the relationship between reproductive success and mating success (the Bateman gradient) is also stronger for males than for females. Sexual selection, based on Bateman's principles, has been demonstrated in animals and some angiosperms, but never in a seaweed. Here we focus on the oogamous haploid-diploid rhodophyte Gracilaria gracilis in which previous studies have shown evidence for non-random mating, suggesting the existence of male-male competition and female choice. We estimated mating and reproductive success using paternity analyses in a natural population where up to 92% of fertilizations occurred between partners of that population. The results show that the variance in mating success is significantly greater in males than in females and that the Bateman gradient is positive only in males. Distance to female partners also explains a minor part of the variance in male mating success. Although there is no evidence for sexual dimorphism, our study supports the hypothesis that sexual selection occurs in G. gracilis, probably on male traits, even if we cannot observe, characterize or quantify them yet.

Newly discovered morphology of the Silurian sea spider Haliestes and its implications

AbstractThe three‐dimensionally preserved Haliestes dasos from the Silurian (Wenlock) Lagerstätte is the most complete fossil sea spider and the oldest unambiguous pycnogonid known from the fossil record. The discovery of two new specimens to add to the holotype reveals new features including proximal annulations of the appendages and segmentation of the trunk end, critical details for comparison with pycnogonids from the Devonian (Emsian) Hunsrück Slate and for the interpretation of the evolutionary significance of Palaeozoic genera. There is some evidence of sexual dimorphism. Haliestes dasos was nektobenthic and its morphology indicates an unusual mode of feeding compared with living pycnogonids. The new morphological features of H. dasos are closely similar to those in Palaeoisopus problematicus from the Hunsrück Slate and it clearly belongs, together with that species, in stem Pycnogonida and not the crown group.

Sex-specific microglia state in the Neuroligin-4 knock-out mouse model of autism spectrum disorder

Neuroligin-4 (NLGN4) loss-of-function mutations are associated with monogenic heritable autism spectrum disorder (ASD) and cause alterations in both synaptic and behavioral phenotypes. Microglia, the resident CNS macrophages, are implicated in ASD development and progression. Here we studied the impact of NLGN4 loss in a mouse model, focusing on microglia phenotype and function in both male and female mice. NLGN4 depletion caused lower microglia density, less ramified morphology, reduced response to injury and purinergic signaling specifically in the hippocampal CA3 region predominantly in male mice. Proteomic analysis revealed disrupted energy metabolism in male microglia and provided further evidence for sexual dimorphism in the ASD associated microglial phenotype. In addition, we observed impaired gamma oscillations in a sex-dependent manner. Lastly, estradiol application in male NLGN4-/- mice restored the altered microglial phenotype and function. Together, these results indicate that loss of NLGN4 affects not only neuronal network activity, but also changes the microglia state in a sex-dependent manner that could be targeted by estradiol treatment.

Associations between maternal dietary scores during early pregnancy with placental outcomes.

Individual macronutrient and micronutrient effects on placental growth have been widely investigated. However, the influence of overall maternal diet is relatively unknown. Therefore, the aim of this study is to examine associations between a range of maternal dietary scores during early pregnancy with placental outcomes, and to investigate whether there is evidence of sexual dimorphism. This analysis of the Lifeways Cross-Generational Cohort includes 276 mother-child pairs. A validated 148-item semi-quantitative food frequency questionnaire assessed maternal diet in early pregnancy. Dietary scores reflecting dietary quality [Healthy Eating Index (HEI-2015), Dietary Approaches to Stop Hypertension (DASH)], dietary inflammatory potential [Dietary Inflammatory Index (DII) and the energy adjusted DII (E-DII)], dietary antioxidant status [Dietary Antioxidant Quality (DAQ)], and glycemic and insulinemic loads/indices (GL/GI, IL/II) were calculated. Linear regression analyses assessed maternal dietary score relationships with untrimmed placental weight (PW) and birth weight:placental weight (BW:PW) ratio. In fully adjusted models, maternal E-DII and GI were positively associated, and HEI-2015 and DAQ were negatively associated with PW (B: 12.31, 95% CI: 0.41, 24.20, p = 0.04, B: 4.13, 95% CI: 0.10, 8.17, p = 0.04, B: -2.70, 95% CI: -5.03, -0.35, p = 0.02 and B: -15.03, 95% CI: -28.08, -1.98, p = 0.02, for E-DII, GI, HEI-2015 and DAQ respectively). Maternal DAQ associations with BW:PW ratio were attenuated. When stratified by sex, maternal GI and pregnancy-specific DAQ were associated with PW in female offspring (B: 5.61, 95% CI: 0.27, 10.96, p = 0.04 and B: -15.31, 95% CI: -30.35, -0.27, p = 0.046). Maternal E-DII and HEI-2015 were associated with PW in males (B: 24.31, 95% CI: 5.66, 42.96, p = 0.01 and B: -3.85, 95% CI: -7.47, -0.35, p = 0.03 respectively). The results of this novel investigation suggest that maternal diet may influence placental development. Female fetuses may be more sensitive to increased glucose levels whereas male fetuses may be more susceptible to in-utero stresses that are regulated by inflammatory pathways and overall diet quality. Hence, early pregnancy offers an opportune time for a mother to prioritize dietary changes that focus on reducing inflammatory and glycemic responses.

Sexual Dimorphism in Brain Sirtuin-1 and m6A Methylated Sirtuin-1 mRNA, and in Protection with Post-Injury Anti-miR-200c treatment, after Experimental Stroke in Aged Mice.

We previously demonstrated that inhibition of miR-200c was protective against stroke in young adult male mice by augmenting sirtuin-1 (Sirt1). In the present study we assessed the role of miR-200c on injury, Sirt1, and bioenergetic and neuroinflammatory markers in aged male and female mice after experimental stroke. Mice were subjected to 1hr of transient middle cerebral artery occlusion (MCAO) and assessed for post-injury expression of miR-200c, Sirt1 protein and mRNA, N6-methyladenosine (m6A) methylated Sirt1 mRNA, ATP, cytochrome C oxidase activity, tumor necrosis factor alpha (TNFα), interleukin-6 (IL-6), infarct volume and motor function. MCAO induced a decrease in Sirt1 expression at 1d post-injury only in males. No differences in SIRT1 mRNA were observed between the sexes. Females had greater baseline miR-200c expression and a greater increase in miR-200c in response to stroke, while pre-MCAO levels of m6A SIRT1 was greater in females. Males had lower post-MCAO ATP levels and cytochrome C oxidase activity, and higher TNFα and IL-6. Post-injury intravenous treatment with anti-miR-200c reduced miR-200c expression in both sexes. In males, anti-miR-200c increased Sirt1 protein expression, reduced infarct volume, and improved neurological score. Conversely in females anti-miR-200c had no effect on Sirt1 levels and provided no protection against injury from MCAO. These results provide the first evidence of sexual dimorphism in the role of a microRNA in aged mice after experimental stroke and suggest sex-differences in epigenetic modulation of the transcriptome and downstream effects on miR biological activity may play a role in sexually dimorphic outcomes after stroke in aged brains.

Fingerprint Patterns in Students with Learning Disability: Evidence of Sexual Dimorphism and Potential for Diagnosis in a Nigerian Population

This study assessed the sex-based relationship and prediction pattern between fingerprint patterns, ridge counts, and learning disability (LD). This cross-sectional study recruited 300 students (150 LD and 150 non-LD) aged between 3 and 29 years. The fingerprint patterns (arch, whorl, ulnar loop, and radial loop) and the ridge count: total finger ridge count (TFRC), absolute ridge count (ARC), ulnar ridge count (URC), and radial ridge count (RRC) were accessed. Students with LD showed a significantly higher whorl and a significantly lower ulnar loop than students without LD. There is a significant association of whorl pattern in the first right finger of subjects with LD compared to non-LD counterparts. TFRC, ARC, and URC were significantly higher in females with LD than non-LD females (P = 0.01, 0.03, and 0.001). Males with LD showed significantly lower TFRC, RRC, and URC counts than the non-LD males (P = 0.02, 0.01, and 0.001). TFRC can predict LD in males (odds ratio [OR] = 1.010, P = 0.032) and females (OR = 0.993, P = 0.012). Fingerprint pattern and ridge counts are sexually dimorphic in subjects with or without LD. TFRC and whorl fingerprint patterns may be vital predictive and screening tools for LD in males and females.

Sexually dimorphic architecture and function of a mechanosensory circuit in C. elegans

How sensory perception is processed by the two sexes of an organism is still only partially understood. Despite some evidence for sexual dimorphism in auditory and olfactory perception, whether touch is sensed in a dimorphic manner has not been addressed. Here we find that the neuronal circuit for tail mechanosensation in C. elegans is wired differently in the two sexes and employs a different combination of sex-shared sensory neurons and interneurons in each sex. Reverse genetic screens uncovered cell- and sex-specific functions of the alpha-tubulin mec-12 and the sodium channel tmc-1 in sensory neurons, and of the glutamate receptors nmr-1 and glr-1 in interneurons, revealing the underlying molecular mechanisms that mediate tail mechanosensation. Moreover, we show that only in males, the sex-shared interneuron AVG is strongly activated by tail mechanical stimulation, and accordingly is crucial for their behavioral response. Importantly, sex reversal experiments demonstrate that the sexual identity of AVG determines both the behavioral output of the mechanosensory response and the molecular pathways controlling it. Our results present extensive sexual dimorphism in a mechanosensory circuit at both the cellular and molecular levels.

Cover Image, Volume 530, Issue 16

The cover image is based on the Research Article Organization of neural systems expressing melanocortin-3 receptors in the mouse brain: Evidence for sexual dimorphism by Michelle N. Bedenbaugh et al., https://doi.org/10.1002/cne.25379. Image Credit: Michelle Bedenbaugh.

Radiographic pelvimetry in free-ranging Eurasian lynx (Lynx lynx carpathicus) from Switzerland

The observation of pelvic anomalies in two Eurasian lynx (subspecies Lynx lynx carpathicus) from a population reintroduced to Switzerland raised the question of the frequency of such anomalies, but no anatomical reference values were available for comparison. This study aimed at providing baseline data on the pelvic morphology of Carpathian lynx from Switzerland, and at detecting potential pelvic anomalies. Measurements of 10 pelvic parameters were performed on the radiographs of 56 lynx taken from 1997–2015. Two ratios (vertical diameter/acetabula; sagittal diameter/transversal diameter) and two areas (pelvic outlet and inlet) were calculated to describe pelvic shape. The results showed that the Eurasian lynx has a mesatipellic pelvis, with a pelvic length corresponding to approximatively 20% of the body length. We found growth-related pelvis size differences among age classes and evidence of sexual dimorphism in adults: two parameters reflecting pelvic width were larger in females, likely to meet the physiological requirements of parturition. By contrast, pelvis length, conjugata vera, diagonal conjugata, sagittal diameter, and tendentially also vertical diameter, were larger in males, in agreement with their larger body size. Outliers were found in five individuals but apparently without clinical significance. Extreme values were likely due to inter-individual differences and the limited sample size rather than to possible congenital or developmental pathological morphology of the pelvic cavity. We present baseline data of the pelvic morphology, including growth and sexual dimorphism, which may be useful for health monitoring and for determination of age and sex in skeletal remains of Carpathian lynx.

Organization of neural systems expressing melanocortin-3 receptors in the mouse brain: Evidence for sexual dimorphism.

The central melanocortin system is fundamentally important for controlling food intake and energy homeostasis. Melanocortin-3 receptor (MC3R) is one of two major receptors of the melanocortin system found in the brain. In contrast to the well-characterized melanocortin-4 receptor (MC4R), little is known regarding the organization of MC3R-expressing neural circuits. To increase our understanding of the intrinsic organization of MC3R neural circuits, identify specific differences between males and females, and gain a neural systems level perspective of this circuitry, we conducted a brain-wide mapping of neurons labeled for MC3R and characterized the distribution of their projections. Analysis revealed MC3R neuronal and terminal labeling in multiple brain regions that control a diverse range of physiological functions and behavioral processes. Notably, dense labeling was observed in the hypothalamus, as well as areas that share considerable connections with the hypothalamus, including the cortex, amygdala, thalamus, and brainstem. Additionally, MC3R neuronal labeling was sexually dimorphic in several areas, including the anteroventral periventricular area, arcuate nucleus, principal nucleus of the bed nucleus of the stria terminalis, and ventral premammillary region. Altogether, anatomical evidence reported here suggests that MC3R has the potential to influence several different classes of motivated behavior that are essential for survival, including ingestive, reproductive, defensive, and arousal behaviors, and is likely to modulate these behaviors differently in males and females.

Tracking sexual dimorphism of facial width-to-height ratio across the lifespan: implications for perceived aggressiveness.

The facial width-to-height ratio (FWHR) influences social judgements like perceived aggression. This may be because FWHR is a sexually dimorphic feature, with males having higher FWHR than females. However, evidence for sexual dimorphism is mixed, little is known about how it varies with age, and the relationship between sexual dimorphism and perceived aggressiveness is unclear. We addressed these gaps by measuring FWHR of 17 607 passport images of male and female faces across the lifespan. We found larger FWHR in males only in young adulthood, aligning with the stage most commonly associated with mate selection and intrasexual competition. However, the direction of dimorphism was reversed after 48 years of age, with females recording larger FWHRs than males. We then examined how natural variation in FWHR affected perceived aggressiveness. The relationship between FWHR and perceived aggressiveness was strongest for males at 27–33 and females at 34–61. Raters were most sensitive to differences in FWHR for young adult male faces, pointing to enhanced sensitivity to FWHR as a cue to aggressiveness. This may reflect a common mechanism for evaluating male aggressiveness from variability in structural (FWHR) and malleable (emotional expression) aspects of the face.

Taxonomic and natural history notes on Oecomys cleberi (Rodentia: Cricetidae) and first records in the Atlantic Forest, Brazil

ABSTRACT The arboreal rodent Oecomys cleberi was previously considered endemic to the Cerrado domain in central-western Brazil. We report on a range extension of O. cleberi to the Atlantic Forest domain of São Paulo, and add new records to Mato Grosso do Sul and Minas Gerais states, totaling 10 new records. Based on a large sample of O. cleberi, we did not find evidence for sexual dimorphism. Our results revealed that specimens of O. cleberi from Cerrado and Atlantic Forest are larger than O. bicolor from the Amazon. Moreover, the larger buccinator–masticatory/foramen ovale confluence in O. cleberi allowed further distinction to O. bicolor. We also provide further details of morphological variation within O. cleberi. Reproductive males and females of O. cleberi were recorded in the rainy season, between December and January, and one male was infested by the mite Gigantolaelaps canestrinii. Our new records show that this arboreal rodent is also present in seasonal forests within the domain of the Atlantic Forest. We suggest that this species, typical of the Cerrado, occurs only marginally in the Atlantic Forest of São Paulo.

Two new species of the genus Gumardee (Marsupialia, Macropodiformes) reveal the repeated evolution of bilophodonty in kangaroos

Two new species of the fossil macropodiform genus Gumardee are described that provide insights into the evolution of early kangaroos. Gumardee has a continuous record from the late Oligocene to the early Miocene and is likely to have become extinct before the middle Miocene. The two new species are the most derived members of the genus. They exhibit some dental size variation, especially in P3/p3, which we interpret as evidence for sexual dimorphism. From the stratigraphically oldest species of Gumardee to the youngest, the dental morphology also shows a progression from bunolophodont to increasingly bilophodont. This trend appears to have occurred in at least four different lineages of macropodoids; however, a fully bilophodont condition was achieved only twice, once in the ancestral lineage of balbarids, and again in ancestral radiation of modern macropodids as implied by the species of Gumardee. Kenny. J. Travouillon [Kenny.Travouillon@museum.wa.gov.au], Western Australian Museum, Locked Bag 49, Welshpool DC, WA, 6986 Australia; Kaylene Butler [kaylene.butler@uqconnect.edu.au ], School of Earth and Environmental Sciences, The University of Queensland, St Lucia, Queensland 4072, Australia; Michael Archer [m.archer@unsw.edu.au ], Earth and Sustainability Science Research Centre, School of Biological, Earth and Environmental Sciences, University of New South Wales, New South Wales 2052, Australia; Suzanne J. Hand [s.hand@unsw.edu.au ], Earth and Sustainability Science Research Centre, School of Biological, Earth and Environmental Sciences, University of New South Wales, New South Wales 2052, Australia.

Ontogenetic skull variation in an Amazonian population of lowland tapir, Tapirus terrestris (Mammalia: Perissodactyla) in the department of Loreto, Peru

ABSTRACT The skulls of 54 specimens of the South American tapir, Tapirus terrestris collected in the department of Loreto, Peru were measured, analyzed and compared to investigate skull development of this species. Univariate, multivariate and allometric analyses were performed using 32 skull variables through traditional morphometrics. Significant skull shape variation was detected among ontogenetic classes. Young individuals (class I, n = 22) showed higher variation than subadults and adults (class II, n = 23 and class III, n = 9), without evidence of sexual dimorphism (males = 35, females = 19). Principal component analyses and discriminant function analysis showed almost complete separation of the age classes. Allometric analysis indicated a tendency of unproportioned cranial growth. All our samples come from the same population living under the same ecological condition, which eliminates the effect of confounding variables related to habitat on the pattern of ontogenetic variation of this anatomical structure.

Sex Differences in the Effect of Diabetes on Cerebral Glucose Metabolism.

The neuroimaging literature indicates that brain structure and function both deteriorate with diabetes, but information on sexual dimorphism in diabetes-related brain alterations is limited. This study aimed to ascertain whether brain metabolism is influenced by sex in an animal model of diabetes. Eleven rats (male, n = 5; female, n = 6) received a single intraperitoneal injection of 70 mg/kg streptozotocin (STZ) to develop diabetes. Another 11 rats (male, n = 5; female, n = 6) received the same amount of solvent through a single intraperitoneal injection. Longitudinal positron emission tomography scans were used to assess cerebral glucose metabolism before and 4 weeks after STZ or solvent administration. Before STZ or solvent injections, there was no evidence of sexual dimorphism in cerebral metabolism (p > 0.05). Compared with healthy control animals, rats with diabetes had significantly decreased brain metabolism in all brain regions (all p < 0.05). In addition, female diabetic rats exhibited further reduction in cerebral metabolism, relative to male diabetic rats (p < 0.05). The results of this study may provide some biological evidence, supporting the existence of a sexual dimorphism in diabetes-related complications.

Prevalence and Characterization of Labial Cervical Vertical Groove in maxillary permanent Incisors: an observational, cross-sectional study

Objective: To evaluate the prevalence and traits of Labial Cervical Vertical Groove in permanent maxillary incisors, its impact on accompanying gingiva, can cause periodontal, esthetic, and prosthetic concern. Aim: The purpose of this study was to determine the frequency and attribute of LCVG in the specific tooth (central vs lateral incisor), side (unilateral vs bilateral, left vs right), and gender. Settings and Design: An in vivo observational cross-sectional study. Materials and Methods: An aggregate of 1800 people aged 18 and 60 were chosen at random and scrutinized for the existence of LCVG in an individual's maxillary permanent incisors. According to predetermined criteria, the deformity was categorized as mild, moderate, or severe. Normal, partial, and irregular gingival covering at the groove site were classified. The inclusion of a Palato-Radial Groove was also detected. Results: LCVG was diagnosed in 123 patients (6.83%). In 59 (47.97%) of the individuals, it was unilateral, while in 64 (52.03%), it was bilateral. There was no evidence of sexual dimorphism or side prevalence. LCVG was identified in 187 (38%) of the 492 teeth in 123 participants. Mild LCVG was detected in 72 incisors, moderate in 83 incisors, and severe in 32 incisors, with a central to lateral incisor ratio of 186:1. In 123 incisors, the gingival contour was normal, partial in 59 incisors, and in 5 incisors it’s irregular. In incisors with moderate or severe LCVG, the gingival recession was noted. Conclusion: An LCVG is a malformation that affects the permanent central incisors of the maxilla. Increased severity of LCVG has deleterious repercussions on the periodontium, necessitating conscientious dental hygiene; accordingly, practitioners should be aware of this, and patients must be advised on the importance of maintaining good oral hygienic practices.