Impact of Lactobacillus crispatus-containing oral and vaginal probiotics on vaginal health: a randomised double-blind placebo controlled clinical trial.

Health of reproductive tract is tightly associated with balance of microbial communities in this area. Bacterial vaginosis (BV) and vulvovaginal candidiasis (VVC) represent common disturbances of vaginal communities. Vaginal discharge due to BV or VVC is a very frequent reason for visiting gynaecologist. We aimed to evaluate the impact of the novel evidence-based probiotics on BV and VVC patients. The study group included 89 BV and 93 VVC patients (aged 18-50 years) who were recruited into randomised double-blind placebo-controlled two-arm parallel trial. The patients of each diagnosis group received oral or vaginal probiotic capsules, or placebo capsules during 3 months. A probiotic capsule contained two (DSM32717 and DSM32720, in case of BV) or three (DSM32720, DSM32718 and DSM32716, in case of VVC) Lactobacillus crispatus strains. Vaginal, intestinal and general health was monitored weekly by questionnaire. Blood analyses were done in the beginning and at the end of trial. Vaginal samples were collected monthly, microscopic and molecular analyses were performed. The study revealed that both oral and vaginal capsules reduced the signs and symptoms in BV patients. Remarkable improvement was noted in Nugent score, amount and smell of discharge, but also in itching/irritation. Consumption of vaginal probiotics significantly increased the lactobacilli counts in their vagina while mean proportion of some BV-related bacteria decreased. In VVC patients, both oral and vaginal capsules lowered the combined score of two most important symptoms, amount of discharge and itching/irritation. In conclusion, the novel formulations of evidence-based well-focused probiotic L. crispatus strains are effective against BV and VVC being suitable for both vaginal and oral administration. Clinical trial registration: ISRCTN34840624, BioMed Central.

Sudden death of a post-partum female with anomalous origin of the coronary artery with bicuspid aortic valve.

Modern techniques have enabled routine diagnosis of congenital cardiac defects, notwithstanding, there will be some that remain undiagnosed and asymptomatic until adulthood. But this is rarely the case with the patent foramen ovale, with the anomalous aortic origin of the left main coronary artery and bicuspid aortic valve. This case describes the sudden death of a female in her post-partum period due to cardiac tamponade following a ruptured aneurysm of the coronary artery at its origin at the sinus of Valsalva. Autopsy shows patent foramen ovale findings and anomalous aortic origin of coronary artery (AAOCA) and bicuspid aortic valve. The association of AAOCA with bicuspid aortic valve and patent foramen ovale is rare.

Dramatically deteriorated quality of life in men with prostatitis-like symptoms.

BACKGROUND: Despite widespread occurrence and poor comprehension, prostatitis has been largely under-researched. OBJECTIVE: To compare complaints, general and sexual health, co-morbidities, risk factors, and lifestyle in men with and without prostatitis-like symptoms (PLS). MATERIAL AND METHODS: The cross-sectional study included 20- to 59-year-old male residents of Estonia. Questionnaire data of 82 men with PLS and of 711 men without PLS were compared. RESULTS AND DISCUSSION: A third of men with PLS considered their health poor, with more frequently diagnosed renal diseases, benign prostate hyperplasia, STDs, chronic nervous system diseases, and depression in them than in controls. They reported more cystitis and gynecological inflammations in their partners, and more prostatitis in their close relatives. This familial predisposition indicates possible genetic and immunologic background of PLS that may be associated also with susceptibility to respiratory tract infections revealed for the first time in our study. By the personality type, the men in the PLS group were less calm but more worrying. Hard drinks, antidepressants, sedative, and sleeping pills were more frequently consumed, and nightshift working and continuous stress were more commonly seen among men with than without PLS. PLS disturbed the sexual life as well as everyday activities. CONCLUSIONS: The men with PLS are characterized by remarkable complex of co-morbidities, habits, and attitudes. PLS possess substantial negative impact on quality of life. Successful work-up of these patients needs multidimensional treatment modalities that take into consideration major factors of syndrome. Genetic factors and central nervous system imbalance but also partner's genital tract microbiota as the potential contributing and/or perpetuating factors to PLS need more scientific attention.

Potential vaginal probiotics: safety, tolerability and preliminary effectiveness.

Vaginal discharge is one of the common reasons for gynaecologist consultation, as bacterial vaginosis and candidiasis are the main causes of discharge. These patients frequently experience numerous problems due to recurrent infections, side effects and drug resistance therefore alternative drugs are needed. Our primary aim was to evaluate safety and tolerability of the potentially probiotic Lactobacillus crispatus strains in volunteer women considering themselves healthy. We also monitored the effects of these strains on vaginal health parameters and lactobacilli counts in vagina and intestine. Forty women were recruited into trial. Absence of chronic diseases was confirmed by questionnaire and blood analysis at screening visit. In randomised double-blind placebo-controlled crossover study the eligible participants were randomly allocated to one of four groups and had to consume one of the two study products (Pro I or Pro II) - a capsule containing 3 strains, 109 cfu per strain, or placebo for 1 week. Treatment period was followed by 2-week washout period and continued with second treatment and washout period. Individuals receiving firstly probiotic, received later placebo and vice versa. Blood, vaginal and faecal samples were collected, and self-reported questionnaires were applied. Thirty subjects completed the trial. The probiotic capsules were well-tolerated. The Pro II intake resulted in a significant decrease in Nugent score (from median 3.0 to 2.0, mean 3.9 to 2.6, P=0.002) and reduction in Gardnerella vaginalis counts (log10 3.57 to 2.38; P=0.027). Reduction of total vaginal bacterial counts was revealed in Pro I group (log10 7.99 to 7.72; P=0.048). In conclusion, the selected vaginal L. crispatus strains are well tolerable and Pro II mixture is prospectively effective in reducing Nugent score and vaginal counts of G. vaginalis. Therefore, these strains seem to be promising candidates for development of novel evidence-based well-focused probiotics to target female urogenital tract disorders.

Impact of sexual debut on culturable human seminal microbiota.

Micro-organisms are tightly integrated into host-microbiota ecosystem. Microbiota of human semen has been studied so far mostly in case of infertility or prostatitis. We aimed to reveal possible impact of sexual debut on seminal microbiota in healthy young men. The study group included 68 young healthy men, of them 12 men without sexual experience, 11 men with single lifetime sexual partner and 45 men with multiple lifetime sexual partners. Basic semen parameters were similar for all subgroups, and no correlation between sexual experience and WBC counts in semen was found. A man could harbour one to nine different bacteria in his semen; the total concentration of bacteria ranged from 2.3 to 7.3 log10 CFU/mL of semen. Lower total bacterial concentration and lower bacterial diversity was observed in men without sexual experience than in sexually experienced men (p < 0.05), with significant positive correlation between these two parameters (r = 0.54; p < 0.0001). In conclusion, the sexual debut is associated with the enrichment of seminal microbiota but not with the influx of WBC or changes in basic seminal parameters.

Suicide by self-disembowelment in prison.

Self-inflicted abdominal injuries are uncommon. Fatal self-inflicted abdominal injury is a rare occurrence, especially as a custodial death. A prisoner under-trial for attempted suicide was in judicial custody and was found unconscious in a pool of blood in the bathroom with cut open abdomen and exposed coils of intestine. The bathroom was found locked from inside and had to be forcibly opened. In spite of immediate hospitalisation and appropriate emergency surgical treatment, he succumbed on the same day. As the person had previously made several futile attempts to commit suicide and was in fact under trial for the crime of attempted suicide (section 309 of Indian Penal Code), the prison authorities had already taken good precautions to prevent access to any type of weapon, instrument, or material that could rekindle the thought of another attempt. In spite of all this, he successfully committed suicide with an apparently innocuous object, a disposable razor. This case is reported for the rarity of occurrence of this type of suicide, and it highlights the importance of utmost precautions to be taken in dealing with prisoners having suicidal tendencies.

Serial murder: An unusual stereotype.

Serial murders attract attention from the media, mental health experts, academia, and the general public. We present a case of serial murders that took place in a limited area and which caused public anxiety and anguish in central India. All the victims were homeless beggars, who were bludgeoned to death (crush injury). Individual murders were initially investigated by different police stations; fortunately, since they sent all the bodies to a common autopsy centre, a forensic pathologist was able to link all the cases, the first person to do so. This emphasises the need for sharing information among police stations and autopsy centres.

Inflammatory reaction found in prostate-specific material - method standardization and proposed optimal cut-off points.

Prostatitis classification as well as treatment decisions are primarily based on differentiation of the inflammatory status in prostate-specific material. At the same time, methods used for detection of inflammation are semi-quantitative and not finally standardized. The main aim of this study was to suggest more precise methods for detection of prostate inflammatory status. Additional aims were to define optimal cut-off points of various tests in order to discriminate between inflammatory and non-inflammatory condition and to analyze the prevalence of inflammatory prostatitis in the groups of symptomatic prostatitis, lower urinary tract symptoms and control subjects. This prospective study included 541 patients (with prostatitis symptoms, with lower urinary tract symptoms and controls) at Tartu University Hospital, Estonia. Leukocyte counts in first-void urine, expressed prostatic secretion and post-massage urine as well as interleukin-6 in prostate secretion specimens were determined. Based on ROC curve analysis, we detected potential normal values for leukocytes in expressed prostatic secretion (<0.5 m/mL), leukocytes in post-massage urine (<4 WBCs per HPF) and IL-6 in prostatic secretion (<50 nmol/mL). Using these newly defined normal values, we detected inflammatory reaction in prostate-specific materials in 24.3%, 53.4% and 69% in control, lower urinary tract symptoms and prostatitis group, respectively. LIMITATION: only subjects with prostatic fluid volume >0.04 mL were included. We propose standardized method for analysis of inflammatory reaction in prostate-specific materials. Based on these newly defined normal values, we verified high prevalence of inflammatory forms of disease in both study groups, but also high prevalence of asymptomatic inflammatory prostatitis among control group of Estonian men. Combining results of analysis of the different inflammatory markers from available prostate-specific materials allows us to differentiate better between inflammatory and non-inflammatory forms of prostate diseases and hence to choose more precise treatment strategies.

Assessment of phenotypic and genotypic antibiotic susceptibility of vaginal Lactobacillus sp.

AIM: To assess antibiotic susceptibility of vaginal lactobacilli strains and provide the data required for assessing the potential of antibiotic resistance risk of new strains selected as probiotic. METHODS AND RESULTS: Potential probiotic vaginal lactobacilli used in the study included 31 vaginal strains of Lactobacillus crispatus (n = 27), Lactobacillus gasseri (n = 3) and Lactobacillus jensenii (n = 1) obtained from the collection of Competence Centre on Health Technologies. Two commercial probiotic strains were used as controls (Lactobacillus rhamnosus GR-1 and Lactobacillus fermentum RC-14). The phenotypic and genotypic antibiotic resistances of the strains were determined by E-test and PCR methods. The location (chromosomal DNA or plasmid) of antibiotic resistance genes was also detected. All lactobacilli strains expressed high level of resistance to kanamycin, metronidazole, norfloxacin and trimethoprim/sulphamethoxazole. Some of the strains also expressed resistance to other antibiotics (chloramphenicol, vancomycin) indicating acquired resistance. I class integrons were found in 20% (6/31) of the strains. The RPP (ribosomal protection protein) gene was found to be positive in 30% (9/31) of the strains. Only one L. jensenii strain was determined with tet(M) gene. The tet(K) gene was positive in 26·7% (8/31) and erm(B) gene in 43·3% (13/31) of strains. Three RPP and both four tet(K) and erm(B) genes were located in plasmids. CONCLUSIONS: High antibiotic resistance to clinically important antibiotics was demonstrated, including metronidazole, sulphonamides, aminoglycoside and quinolones. In addition, acquired tetracycline and erythromycin resistance genes were detected in either plasmid or chromosomal DNA of certain isolates, in some of the cases for the first time in the literature. It appears that antibiotic resistance genes erm(B) and tet(K) are widely spread in vaginal lactobacilli. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new data about antimicrobial resistance and genotypic diversity of vaginal Lactobacillus isolates. In addition, it provides data assessing the potential of antibiotic resistance risk of new strains selected as probiotic.

Couple-related factors of ART outcome.

PURPOSE OF INVESTIGATION: Although assisted reproductive technologies (ART) are the current mainstay of infertility treatment, several mechanisms leading to ART failure are still unclear. The aim of the present study was to determine possible health-related risk factors in both counterparts of the couple affecting the ART outcome. MATERIAL AND METHODS: The authors recruited 100 consecutive couples who were undergoing ART procedure. All subjects filled out a health questionnaire, and the presence of bacterial vaginosis in females and/or prostatitis in males was evaluated. RESULTS: Success of the ART procedure was 30.2%, failure being most significantly related to woman's age (OR = 0.24, 95% CI 0.09-0.66, p = 0.003) and excess body weight (OR = 0.08, 95% CI 0.01-0.63, p = 0.002). Male factors, previous conceptions, and presence of children increased the likelihood of ART success. Bacterial vaginosis and prostatitis did not significantly compromise the effectiveness of the ART procedure in this study. CONCLUSION: In 100 consecutive infertile couples scheduled for ART procedure, pregnancy was achieved in 30.2%, with female age and overweight and previous children from male side being the most significant factors affecting the ART outcome.

Prospects of poisoning - a multi facet study.

Aim of the study is to find out demographic profile, clinical characteristics and analysis of poison in clinical set up. The study carried out in Sri Aurobindo Medical College and PG Institute Indore, Madhya Pradesh. Total 75 cases of poisoning were studied for demographic profile, vitals (BP, pulse, heart rate, pupils, etc.), clinical features (such as vomiting, salivation, consciousness, etc.), type of poison and its analysis. Results: Poisoning was more common in cases between 15 and 25 years of age, in males than in females and in Hindu religion. Poisoning cases were predominantly from rural areas and in married people. Majority of cases were discharged after proper treatment and counseling. Altered vitals and clinical features were found in most of the cases. Organophosphate and aluminum phosphide compound were evaluated in most of the cases. Conclusions: Preventive measures should be applied through educating people, proper counseling, promoting poison information centers, and introducing separate toxicological units in hospitals.

Comparison of detection methods for vaginal lactobacilli.

Vaginal lactobacilli offer protection against microbiota imbalance and genitourinary tract infections. We compared vaginal lactobacilli in 50 Estonian women of child-bearing age applying culture-based methods, quantitative PCR and next-generation sequencing (NGS). The culture-based methods found three different lactobacilli: Lactobacillus crispatus, Lactobacillus jensenii and Lactobacillus gasseri. Using NGS revealed the presence of L. crispatus in 76%, Lactobacillus iners in 52%, L. jensenii in 47% and L. gasseri in 33% of the samples. According to qPCR, L. iners was present in 67% and L. crispatus in 64% of the samples. The proportions of L. crispatus revealed by qPCR and NGS were in good correlation (R=0.79, P<0.001), while that of L. iners correlated poorly (R=0.13, P>0.05). Good concordance for L. crispatus was also found between the results of the culture-based method and qPCR. Finally, good overlap between the results of the culture-based method and NGS was revealed: in case of a positive NGS result for L. crispatus, the same species was isolated in 95% of samples. The corresponding percentages were 82% for L. jensenii and 86% for L. gasseri. Our data indicate fairly general concordance of the three methods for detecting vaginal lactobacilli, except for L. iners. This points out the importance of standardisation of techniques, and the respective studies should involve cultures applying a medium suitable for the fastidious L. iners.

Systemic oxidative stress could predict assisted reproductive technique outcome.

PURPOSE: Previous studies have indicated that OxS (oxidative stress) may appear as a possible reason for poor ART outcome. Our aim was to study OxS levels in both partners of couples seeking Assisted reproduction Technology (ART). METHODS: Altogether 79 couples were recruited. Oxidative DNA damage (8-OHdG) and lipid peroxidation (8-EPI) were measured, and clinical background and ART outcomes were recorded. RESULTS: Both OxS markers accurately reflected clinical conditions with prominent negative effects attributable to genital tract infections, endometriosis, uterine myoma and smoking. Furthermore, the level of OxS was also affected by partner's state of health. The highest 8-EPI levels were detected in both partners when biochemically detectable pregnancies did not develop into clinically detectable pregnancies (in women, 97,8 ± 16,7 vs 72.9 ± 22,9, p = 0.007; in men, 89.6 ± 20,4 vs 72,1 ± 22,6, p = 0.049). CONCLUSIONS: To conclude, high grade systemix OxS in both partners may negatively affect the maintenance and outcome of pregnancy. Applying the detection of OxS in ART patients may select patients with higher success rate and/or those who require antioxidant therapy. This would lead to improvement of ART outcome as well as natural fertility.

Neuroprotective and nootropic activity of Clitorea ternatea Linn.(Fabaceae) leaves on diabetes induced cognitive decline in experimental animals.

PURPOSE: Ethanol extract of Clitorea ternatea (EECT) was evaluated in diabetes-induced cognitive decline rat model for its nootropic and neuroprotective activity. MATERIALS AND METHODS: Effect on spatial working memory, spatial reference memory and spatial working-reference memory was evaluated by Y maze, Morris water maze and Radial arm maze respectively. Neuroprotective effects of EECT was studied by assaying acetylcholinesterase, lipid peroxide, superoxide dismutase (SOD), total nitric oxide (NO), catalase (CAT) and glutathione (GSH) levels in the brain of diabetic rats. RESULTS: The EECT (200 and 400 mg/kg) was found to cause significant increase in spatial working memory (P < 0.05), spatial reference memory (P < 0.001) and spatial working-reference (P < 0.001) in retention trials on Y maze, Morris water maze and Radial arm maze respectively. Whereas significant decrease in acetylcholinesterase activity (P < 0.05), lipid peroxide (P < 0.001), total NO (P < 0.001) and significant increase in SOD, CAT and GSH levels was observed in animals treated with EECT (200 and 400 mg/kg) compared to diabetic control group. CONCLUSIONS: The present data indicates that Clitorea ternatea tenders protection against diabetes induced cognitive decline and merits the need for further studies to elucidate its mode of action.

Reproductive function in middle-aged males: healthy men versus male partners of infertile couples.

The aim of this study was to compare the reproductive parameters and the health-related, lifestyle and educational factors in middle-aged healthy men and male partners of infertile couples. Our patient group included 164 male partners of infertile couples with a preceding period of infertility of at least 12 months and 61 men attending a prostate health screening and considering themselves healthy. Significant differences between the groups were found in testicular volume, total sperm output, density and morphology as well as oestradiol levels in serum. The analysis showed significant positive correlations between testicular volume and semen quality, while negative correlations were observed between gonadotrophin levels and sperm parameters in both groups. Physical activity and sexual capability were higher in healthy men, while coital frequency and a history of sexually transmitted diseases were higher in male partners of infertile couples. The impact of physical activity and sexual capability on semen quality for all subjects was revealed. We can conclude that impaired reproductive function, that is, semen quality, in men >45 years is related not only with general male ageing but obvious differences between subjects of infertile couples and healthy middle-aged men can be seen. Their relations with lifestyle, environmental or physiological factors need further study.

Increased levels of hydrogen peroxide and nitric oxide in male partners of infertile couples.

We investigated the prevalence of oxidative stress in male partners of infertile couples displaying different inflammation patterns in their genital tract and/or oligospermia. The levels of acknowledged oxidative stress markers (8-isoprostanes [8-EPI], diene conjugates, reactive oxygen species-total antioxidant capacity [ROS-TAC] score) were elevated in both leukocytospermic men and subjects whose inflammation was limited only to expressed prostatic secretion and/or post-massage urine. Oligospermic men with severe inflammation were different from oligospermic men who had no inflammation at all - the former had elevated 8-EPI, diene conjugates and ROS-TAC score when compared to the latter indicating that inflammation has substantially more significant impact on oxidative stress markers than oligospermia status. At the same time nitric oxide (NO) and hydrogen peroxide (H2 O2 ) levels were significantly increased not only in the men with severe inflammation but also in men with borderline inflammation in their genital tact and in men having non-inflammatory oligospermia. NO, H2 O2 and 8-EPI were negatively correlated with per cent of normal sperms, and NO and H2 O2 showed negative correlation also with sperm count. We can conclude that in men presenting with couple infertility the acknowledged oxidative stress markers are substantially associated with markers of inflammation in genital tract while NO and H2 O2 display high levels also in patients with mild inflammation and non-inflammatory oligospermia.

Sexual intercourse with leukocytospermic men may be a possible booster of oxidative stress in female partners of infertile couples.

Human semen has undoubtedly significant influence on the organism of the female counterpart. At the same time there are no studies in English literature investigating the influence of sexual intercourse on oxidative stress level in women's organism. Seventeen infertile couples where male partners were with (n = 5) or without (n = 12) leukocytospermia were enrolled in the study. Systemic oxidative stress levels were measured, whereby twice in female partners - before and 8-12 h after sexual intercourse. The men with leukocytospermia were characterized by oxidative stress that was substantially transferred to their partners during sexual intercourse as revealed by increase in 8-isoprostanes level (median 32.7 vs. 70.4 ng/mmol creatinine, p = 0.006). Sexual intercourse with male partner having leukocytospermia increases the oxidative stress level in the women's organism that may interfere with fertilization.

Antihyperglycemic and antioxidant activity of Clitorea ternatea Linn. on streptozotocin-induced diabetic rats.

Ethanol extract of Clitorea ternatea Linn. (EECT) was evaluated for its antihyperglycemic and antioxidative activity in normal and streptozotocin-induced diabetic rats. Antihyperglycemic activity of EECT was studied in normal fasted and glucose fed hyperglycemic and epinephrine induced hyperglycemic rats by estimating fasting serum glucose (FSG) by glucose oxidisae or peroxidase enzymatic method. Antioxidant activity of EECT was studied by assaying lipid peroxide/Thiobarbituric acid reactive substances (TBARS), superoxide dismutase (SOD), total nitric oxide, catalase (CAT) and glutathione levels in diabetic rats. The EECT (200 and 400 mg/kg) showed significant antihyperglycemic activity by decreasing FSG in all hyperglycemic models except epinephrine induced hyperglycemic rats; in which improvement in FSG was observed only with EECT in 400 mg/kg dose, whereas significant decrease in TBARS (P < 0.001), nitric oxide (P < 0.001) and significant increase in SOD (P < 0.001), CAT (P < 0.01) and reduced glutathione levels (P < 0.001) was observed in animals treated with EECT (200 and 400 mg/kg) compared to diabetic control group. The results indicated that EECT has remedial effects on hyperglycemia and oxidative stress in diabetic rats.

Clitorea ternatea, a herb from Indian folklore, improves streptozotocin-induced diabetes and diabetes-induced cognitive decline in rats.

OBJECTIVE: To study the antidiabetic, neurochemical-antioxidant and cognition protective effects of Clitorea ternatea leaves on a rat model of diabetic cognitive decline. METHODS: Antidiabetic activity was evaluated by serum glucose and body weight estimation in ethanol extract of Clitorea ternatea (EECT)-treated diabetic rats. Effects of EECT on spatial working memory (SWM) and spatial reference memory (SRM) were evaluated by Y-maze and Morris water maze tests respectively. Neurochemical-antioxidant effects of EECT were studied by acetylcholinesterase assay, and measurements of thiobarbituric acid reactive substances (TBARSs), superoxide dismutase (SOD) and catalase (CAT) levels in diabetic rats. RESULTS: The 200 and 400 mg/kg of EECT showed a significant antidiabetic activity by decreasing serum glucose level (P<0.05, P<0.01), and there was a significant increase in the body weight in 400 mg/kg of EECT-treated diabetic rats (P<0.01). EECT was found to cause significant increases in SWM and SRM in retention trials on Y-maze and Morris water maze respectively (P<0.05, P<0.01). Significant decreases in acetylcholinesterase activity and TBARS level, and significant increase in CAT level were observed in rats treated with 200 and 400 mg/kg of EECT compared with rats in the diabetic control group (P<0.05 or P<0.01). Significant increase was also found in SOD in rats treated with 400 mg/kg of EECT. CONCLUSION: Clitorea ternatea exhibits antidiabetic and antioxidant activities, offers the protection against diabetes-induced cognitive decline, and warrants the need for further studies to elucidate its mode of action.

Characterization of ergocalciferol loaded solid lipid nanoparticles.

UNLABELLED: The use of solid lipid nanoparticles (SLNs) is a technique that has been widely used in the pharmaceutical industry for the last 2 decades and has become of increasing interest to food scientists due to its potential for encapsulation and controlled release. Ergocalciferol (vitamin D2) is a bioactive compound whose deficiency may lead to rickets in children and osteomalacia in adults. In this study, ergocalciferol was encapsulated in tripalmitin SLNs stabilized by polysorbate 20 (Tween 20). SLN dispersions (5% w/w) were prepared by hot homogenization technique using a nozzle-type high-pressure homogenizer. Ergocalciferol at 0%, 5%, 10%, 15%, and 20% (w/w of lipid) was dissolved in the molten lipid at 80 °C, mixed with a 5% (w/w) aqueous solution of polysorbate 20 and homogenized at 138 MPa at 80 °C. Particle size, thermal properties, and microstructure were evaluated by dynamic light scattering (DLS), differential scanning calorimetry (DSC), and transmission electron microscopy (TEM) respectively. As the proportion of ergocalciferol in the SLN increased from 0% to 20%, the Z-average values of the particles gradually decreased (P≤ 0.05) from approximately 120 nm to approximately 65 nm. DSC analysis of freeze dried SLN samples showed gradual decrease in enthalpies of fusion and crystallization for stable ß-subcell whereas for SLN dispersions, the enthalpy of fusion of unstable α-subcell crystal increased with increased ergocalciferol loading. The TEM images of the ergocalciferol loaded SLN samples showed the presence of spherical as well as rod-shaped nanoparticles. It was also observed that the turbidity of the SLN dispersions reduced noticeably with increased ergocalciferol loading. This finding could be useful in terms of fortification of clear juices with ergocalciferol. PRACTICAL APPLICATION: Solid lipid nanoparticles (SLNs) were used in this study to encapsulate vitamin D2, a vitamin important for bone health. It was found that as the concentration of vitamin D2 increased in the lipid phase of SLN dispersion, the clarity of the dispersion increased. Also, with increased vitamin D2 concentration, the stability of lipid crystal structure was affected in a way that indicates higher capacity of lipid to incorporate the vitamin molecules and hence to protect them better from oxygen and light. This vitamin loaded SLNs may offer alternatives to milk and margarine as a source of vitamin D.