Currently there is no treatment for juvenile Batten disease, a fatal childhood neurodegenerative disorder caused by mutations in the CLN3 gene. The Cln3-knockout (Cln3Δex1-6) mouse model recapitulates several features of the human disorder. Cln3Δex1-6 mice, similarly to juvenile Batten disease patients, have a motor coordination deficit detectable as early as postnatal day 14. Previous studies demonstrated that acute attenuation of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)-type glutamate receptor activity by the non-competitive AMPA antagonist, EGIS-8332, in both 1- and 6–7-month-old Cln3Δex1-6 mice results in improvement in motor coordination. Here we show that acute inhibition of N-methyl-D-aspartate (NMDA)-type glutamate receptors by memantine (1 and 5 mg/kg i.p.) had no effect on the impaired motor coordination of one-month-old Cln3Δex1-6 mice. At a later stage of the disease, in 6–7-month-old Cln3Δex1-6 mice, memantine induced a delayed but extended (8 days) improvement of motor skills similarly to that observed previously with EGIS-8332 treatment. An age-dependent therapeutic effect of memantine implies that the pathomechanism in juvenile Batten disease changes during disease progression. In contrast to acute treatment, repeated administration of memantine or EGIS-8332 (1 mg/kg, once a week for 4 weeks) to 6-month-old Cln3Δex1-6 mice had no beneficial effect on motor coordination. Moreover, repeated treatments did not impact microglial activation or the survival of vulnerable neuron populations. Memantine did not affect astrocytosis in the cortex. EGIS-8332, however, decreased astrocytic activation in the somatosensory barrelfield cortex.
Acute inhibition of NMDA receptors can induce a prolonged therapeutic effect, identifying NMDA receptors as a new therapeutic target for juvenile Batten disease.
juvenile Batten disease; neuronal ceroid lipofuscinoses; Cln3; NMDA receptor; AMPA receptor; rotarod
•Macrophages can internalise LDL through scavenger receptor-independent mechanisms.•Macropinocytosis has been shown to contribute significantly to foam cell formation.•Cytokines such as TGF-β, IL-33, IFN-γ and IL-17A can modulate macropinocytosis.•TGF-β mediated inhibition of macropinocytosis is a Smad-2/-3-independent process.•Macropinocytosis is a promising target for therapeutic intervention of atherosclerosis.
A key event during the formation of lipid-rich foam cells during the progression of atherosclerosis is the uptake of modified low-density lipoproteins (LDL) by macrophages in response to atherogenic mediators in the arterial intima. In addition to scavenger receptor-dependent uptake of LDL, macropinocytosis is known to facilitate the uptake of LDL through the constitutive and passive internalization of large quantities of extracellular solute. In this study we confirm the ability of macropinocytosis to facilitate the uptake of modified LDL by human macrophages and show its modulation by TGF-β, IFN-γ, IL-17A and IL-33. Furthermore we show that the TGF-β-mediated inhibition of macropinocytosis is a Smad-2/-3-independent process.
AcLDL, acetylated LDL; Apo, apolipoprotein; BMDM, bone marrow-derived macrophage; CD-36, cluster of differentiation 36; DiI, 1,1-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyane perchlorate; THP-1, human acute monocytic leukemia cells; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HMDM, human monocyte-derived macrophages; IL, interleukin; IFN-γ, interferon-γ; LDL, low-density lipoproteins; LY, lucifer yellow; OxLDL, oxidized LDL; SR-A, scavenger receptor A; shRNA, short hairpin RNA; TGF-β, transforming growth factor-β; Macropinocytosis; Cytokine; Foam cell; Atherosclerosis; Low density lipoprotein
•Atherosclerosis is an inflammatory disorder regulated by cytokines.•ADAMTS proteases have been suggested to play an important role in this disease.•The action of key cytokines on the expression of ADAMTS proteases in macrophages is poorly understood.•The effect of IFN-γ, TGF-β, TL1A and IL-17A on the expression of ADAMTS-1, -4 and -5 was studied.•Novel differential actions and synergistic interactions were identified.
Atherosclerosis is an inflammatory disease of the vasculature regulated by cytokines. Macrophages play a crucial role at all stages of this disease, including regulation of foam cell formation, the inflammatory response and stability of atherosclerotic plaques. For example, matrix metalloproteinases produced by macrophages play an important role in modulating plaque stability. More recently, the ADAMTS proteases, which are known to play a key role in the control of cartilage degradation during arthritis, have been found to be expressed in atherosclerotic lesions and suggested to have potentially important functions in the control of plaque stability. Unfortunately, the action of cytokines on the expression of ADAMTS family in macrophages is poorly understood. We have investigated the effect of classical cytokines (IFN-γ and TGF-β) and those that have been recently identified (TL1A and IL-17) on the expression of ADAMTS-1, -4 and -5 in human macrophages. The expression of all three ADAMTS members was induced during differentiation of monocytes into macrophages. TGF-β had a differential action with induction of ADAMTS-1 and -5 expression and attenuation in the levels of ADAMTS-4. In contrast, IFN-γ suppressed the expression of ADAMTS-1 without having an effect on ADAMTS-4 and -5. Although TL-1A or IL-17A alone had little effect on the expression of all the members, they induced their expression synergistically when present together. These studies provide new insight into the regulation of key ADAMTS family members in human macrophages by major cytokines in relation to atherosclerosis.
ADAMTS, a disintegrin and metalloproteinase with thrombospondin motifs; ApoB, apolipoprotein B; ApoE, apolipoprotein E; DR3, death receptor 3; ECM, extracellular matrix; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HMDM, human monocyte-derived macrophages; IFN-γ, interferon-γ; IL, interleukin; LDL, low-density lipoprotein; LDLR, LDL receptor; LPL, lipoprotein lipase; MMP, matrix metalloproteinase; PMA, phorbol 12-myristate 13-acetate; RT-qPCR, real-time quantitative polymerase chain reaction; TGF-β, transforming growth factor-β; TL1A, tumour necrosis factor-like protein 1A; TNF-α, tumour necrosis factor-α; VSMC, vascular smooth muscle cells; ADAMTS proteases; Atherosclerosis; Cytokines; Macrophages; Gene expression
A case of multidrug resistance central nervous system tuberculosis is described. During the initial 6 months of therapy, intracranial embolic spread of tuberculomas from exudates around the proximal left middle cerebral artery was seen. This phenomenon was reported earlier. Further management, therapeutic considerations, particularly with secondary and tertiary line of antitubercular medication and neuroimaging are discussed.
Carotid intima-media thickness (CIMT) and carotid plaques represent preclinical markers of atherosclerosis. We sought to describe predictors of CIMT and carotid plaques, including early life growth, in a young urban Indian cohort free of clinical cardiovascular disease (CVD).
In 2006-2009, we performed B-mode carotid ultrasound on 600 participants (mean [SD] age 36 [1.1] years; 45% women) from the New Delhi Birth Cohort to evaluate CIMT and carotid plaques (> 1mm). Height and weight were recorded at birth, 2 and 11 years of age. Data on CVD risk factors, anthropometry, medical history, socio-economic position, and lifestyle habits were collected in 1998-2002.
Mean (SD) CIMT for men and women was 0.91 (0.12) and 0.86 (0.13) mm, respectively. Carotid plaque was present in 33% of men and 26% of women. Waist circumference in 1998-2002 was positively associated with CIMT (β coefficient 0.26 mm [0.17, 0.36] per SD) and carotid plaque (OR 1.27 [1.06,1.52] per SD) in 2006-2009. Higher triglycerides, PAI-1, insulin resistance, and diastolic blood pressure, metabolic syndrome, and lower HDL-cholesterol and physical activity predicted higher CIMT and/or plaque (p<0.05). Longer length at 2 years was associated with higher CIMT (p<0.05). These associations were attenuated after adjusting for adult waist circumference.
These are the first prospective data from India showing that early life growth, adult socio-demographics, and CVD risk factors predict future CIMT and /or carotid plaque. These relationships appear primarily mediated through central adiposity, highlighting the importance of maintaining a healthy weight in early adulthood to prevent CVD.
carotid intima media thickness; carotid plaque; India; cohort; birth weight; infant and childhood growth
A number of environmental factors (e.g. toxicants) have been shown to promote the epigenetic transgenerational inheritance of disease and phenotypic variation. Transgenerational inheritance requires the germline transmission of altered epigenetic information between generations in the absence of direct environmental exposures. The primary periods for epigenetic programming of the germ line are those associated with primordial germ cell development and subsequent fetal germline development. The current study examined the actions of an agricultural fungicide vinclozolin on gestating female (F0 generation) progeny in regards to the primordial germ cell (PGC) epigenetic reprogramming of the F3 generation (i.e. great-grandchildren). The F3 generation germline transcriptome and epigenome (DNA methylation) were altered transgenerationally. Interestingly, disruptions in DNA methylation patterns and altered transcriptomes were distinct between germ cells at the onset of gonadal sex determination at embryonic day 13 (E13) and after cord formation in the testis at embryonic day 16 (E16). A larger number of DNA methylation abnormalities (epimutations) and transcriptional alterations were observed in the E13 germ cells than in the E16 germ cells. These observations indicate that altered transgenerational epigenetic reprogramming and function of the male germline is a component of vinclozolin induced epigenetic transgenerational inheritance of disease. Insights into the molecular control of germline transmitted epigenetic inheritance are provided.
Thrombocytopenia in pregnancy carries a major risk of feto-maternal morbidity and mortality. We present a case of hypocellular bone marrow with severe thrombocytopenia with pregnancy induced hypertension (PIH) for emergency lower segment cesarean section (LSCS). This disease is characterized by pancytopenia and hypocellular bone marrow with impaired morphology and maturation. Causes of death due to this disease include hemorrhage and infection secondary to thrombocytopenia and neutropenia especially following surgery. We report successful management of emergency LSCS with severe thrombocytopenia with severe PIH.
Hypocellular bone marrow; pregnancy induced hypertension; thrombocytopenia
Chronic hepatitis C remains a significant medical and economic burden in Canada, affecting nearly 1% of the population. Since the last consensus conference on the management of chronic hepatitis C, major advances have warranted a review of recommended management approaches for these patients. Specifically, direct-acting antiviral agents with dramatically improved rates of virological clearance compared with standard therapy have been developed, and several single nucleotide polymorphisms associated with an increased probability of spontaneous and treatment-induced viral clearance have been identified. In light of this new evidence, a consensus development conference was held in November 2011; the present document highlights the results of the presentations and discussions surrounding these issues. It reviews the epidemiology of hepatitis C in Canada, preferred diagnostic testing approaches and recommendations for the treatment of chronically infected patients with the newly approved protease inhibitors (boceprevir and telaprevir), including those who have previously failed pegylated interferon and ribavirin therapy. In addition, recommendations are made regarding approaches to reducing the burden of hepatitis C in Canada.
Antiviral; Boceprevir; Guideline; Hepatitis C; Interferon; Peginterferon; Protease inhibitor; Ribavirin; Telaprevir; Therapy; Treatment
To study the relationship of height and body mass index (BMI) during childhood with adult bone mineral content (BMC), areal density (aBMD) and apparent density (BMAD, estimated volumetric density).
Participants were 565 men and women aged 33-39 years from the New Delhi Birth Cohort, India, whose weight and height were recorded at birth and annually during infancy (0-2 years), childhood (2-11 years) and adolescence (11 years-adult). Lumbar spine, femoral neck and forearm BMC and aBMD were measured using dual X-ray absorptiometry; lumbar spine and femoral neck BMAD were calculated.
Birth length, and height and height gain during infancy, childhood and adolescence were positively correlated with adult BMC (p≤0.01 all sites except birth length with femoral neck). Correlations increased with height from birth-6 years, then remained constant for later height measurements. There were no associations with BMAD. BMI at birth, and during childhood and adolescence was also positively correlated with BMC (p<0.01 all sites). BMI at 11 years, and BMI gain in childhood and adolescence, were correlated with aBMD and BMAD (p<0.001 for all); these correlations strengthened with increasing age of BMI measurement. The associations with height and BMI in early life became non-significant after adjustment for adult height and BMI.
Greater skeletal growth and BMI gain in utero and during infancy are associated with higher peak BMC, and greater BMI gain in childhood and adolescence is associated with higher peak aBMD and BMAD. These associations are mediated by the attainment of adult height and BMI respectively.
Birth cohort; childhood growth; height; body mass index; bone mineral content; bone mineral density
Leprosy involves peripheral nerves sooner or later in the course of the disease leading to gross deformities and disabilities. Sadly, by the time it becomes clinically apparent, the nerve damage is already quite advanced. However, if the preclinical damage is detected early in the course of disease, it can be prevented to a large extent.
Materials and Methods:
We conducted an electrophysiological pilot study on 10 patients with clinically manifest leprosy, in the Dermatology Department of Mahatma Gandhi Institute of Medical Sciences, Sewagram. This study was done to assess the nerve conduction velocity, amplitude and latency of ulnar and median nerves.
Results and Conclusion:
We found reduced conduction velocities besides changes in latency and amplitude in the affected nerves. Changes in sensory nerve conduction were more pronounced. Also, sensory latencies and amplitude changes were more severe than motor latencies and amplitude in those presenting with muscle palsies. However, further studies are going on to identify parameters to detect early nerve damage in leprosy.
Electrophysiology; leprosy; nerve conduction study
Saccharum officinarum is one of the most cultivated hybrid varieties among the sugarcane varieties. In sugarcane plant sucrose is the
major carbohydrate which can be stored and transported. Different physiological and biochemical studies on this crop report that
invertase activity and sucrose concentration some how are key limiting step in the process of sucrose accumulation. Significant
efforts have been made in relation to the sucrose cycle by altering the sucrose phosphate synthetase, sucrose synthetase and
invertase. In sugarcane two types of invertase enzymes have been reported on the basis of pH and cellular localization. Invertase
breaks the sucrose into hexoses as a source of energy and carbon. It has also been reported that this enzyme is involved in the
process of cell differentiation and plant development. Progress has been made for the understanding of invertase activity and its
role in sugarcane plant. With the help of biotechnology it is possible to target the desired gene with genetic engineering approach
to increase sucrose content by careful manipulation of invertase (enzyme) gene to increase the sucrose yield in sugarcane. Purpose
of this mini review is to high-light the role of invertase in sugarcane and how to overcome sucrose recovery in sugarcane.
Sugarcane; Invertase; Sucrose metabolism; Differentiation; Development
This study considers three questions: 1. What are the Canadian public’s prioritization preferences for new government spending on a range of public health-related goods outside the scope of the country’s national system of health insurance? 2. How homogenous or heterogeneous is the Canadian public in terms of these preferences? 3. What factors are predictive of the Canadian public’s preferences for new government spending? Data were collected in 2008 from a national random sample of Canadian adults through a telephone interview survey (n =1,005). Respondents were asked to rank five spending priorities in terms of their preference for new government spending. Bivariate and multivariable logistic regression analyses were conducted. As a first priority, Canadian adults prefer spending on child care (26.2%), followed by pharmacare (23.1%), dental care (20.8%), home care (17.2%), and vision care (12.7%). Sociodemographic characteristics predict spending preferences, based on the social position and needs of respondents. Policy leaders need to give fair consideration to public preferences in priority setting approaches in order to ensure that public health-related goods are distributed in a manner that best suits population needs.
Public preferences; Health care; Priority setting; Health services needs and demand
The anti-atherogenic cytokine, TGF-β, plays a key role during macrophage foam cell formation by modulating the expression of key genes involved in the control of cholesterol homeostasis. Unfortunately, the molecular mechanisms underlying these actions of TGF-β remain poorly understood. In this study we examine the effect of TGF-β on macrophage cholesterol homeostasis and delineate the role of Smads-2 and ‐3 during this process. Western blot analysis showed that TGF-β induces a rapid phosphorylation-dependent activation of Smad-2 and ‐3 in THP-1 and primary human monocyte-derived macrophages. Small interfering RNA-mediated knockdown of Smad-2/3 expression showed that the TGF-β-mediated regulation of key genes implicated in the uptake of modified low density lipoproteins and the efflux of cholesterol from foam cells was Smad-dependent. Additionally, through the use of virally delivered Smad-2 and/or Smad-3 short hairpin RNA, we demonstrate that TGF-β inhibits the uptake of modified LDL by macrophages through a Smad-dependent mechanism and that the TGF-β-mediated regulation of CD36, lipoprotein lipase and scavenger receptor-A gene expression was dependent on Smad-2. These studies reveal a crucial role for Smad signaling, particularly Smad-2, in the inhibition of foam cell formation by TGF-β through the regulation of expression of key genes involved in the control of macrophage cholesterol homeostasis.
► Anti-atherogenic cytokine TGF-β inhibits macrophage foam cell formation. ► The role of Smads in the control of macrophage cholesterol homeostasis was studied. ► Smads were found to play a key role in the TGF-β-mediated uptake of modified LDL. ► A dominant role of Smad2 was identified in the regulation of gene expression. ► The TGF-β-Smad axis may represent a powerful anti-foam cell therapeutic target.
AcLDL, acetylated low density lipoprotein; ABCA-1, ATP-binding cassette transporter A-1; ABCG-1, ATP-binding cassette transporter G-1; ApoE, apolipoprotein E; ApoE−/−, apolipoprotein E deficient; CD36, cluster of differentiation 36; DiI, 1,1′-dioctadecyl-3,3,3′,3′-tetramethyllindocarbocyane perchlorate; HMDM, human monocyte-derived macrophages; LDL, low density lipoprotein; LPL, lipoprotein lipase; OxLDL, oxidized low density lipoprotein; shRNA, short hairpin RNA; SR-A, scavenger receptor A; THP-1, human acute monocytic leukemia cell line; Foam cell; Atherosclerosis; Cholesterol; TGF-β; Macrophage
A major event in mammalian male sex determination is the induction of the testis determining factor Sry and its downstream gene Sox9. The current study provides one of the first genome wide analyses of the downstream gene binding targets for SRY and SOX9 to help elucidate the molecular control of Sertoli cell differentiation and testis development. A modified ChIP-Chip analysis using a comparative hybridization was used to identify 71 direct downstream binding targets for SRY and 109 binding targets for SOX9. Interestingly, only 5 gene targets overlapped between SRY and SOX9. In addition to the direct response element binding gene targets, a large number of atypical binding gene targets were identified for both SRY and SOX9. Bioinformatic analysis of the downstream binding targets identified gene networks and cellular pathways potentially involved in the induction of Sertoli cell differentiation and testis development. The specific DNA sequence binding site motifs for both SRY and SOX9 were identified. Observations provide insights into the molecular control of male gonadal sex determination.
Weight gain and growth in early life may influence adult pro-inflammatory and pro-thrombotic cardiovascular risk factors.
Follow-up of a birth cohort in New Delhi, India, whose weight and height were measured 6-monthly until age 21 years. BMI at birth, during infancy (2 years), childhood (11 years) and adulthood (26-32 years) and BMI gain between these ages were analyzed in 886 men and 640 women in relation to adult fibrinogen, high-sensitivity C-reactive protein (hsCRP) and plasminogen activator inhibitor (PAI-1) concentrations.
All the pro-inflammatory/pro-thrombotic risk factors were higher in participants with higher adiposity. In women, BMI at birth and age 2 years was inversely related to fibrinogen (p=0.002 and 0.05) and, after adjusting for adult adiposity, to hsCRP (p=0.02 and 0.009). After adjusting for adult adiposity, BMI at 2 years was inversely related to hsCRP and PAI-1 concentrations (p<0.001 and p=0.02) in men. BMI gain between 2-11 years and/or 11 years to adulthood was positively associated with fibrinogen and hsCRP in women and with hsCRP and PAI-1 in men.
Thinness at birth or during infancy, and accelerated BMI gain during childhood/adolescence are associated with a pro-inflammatory/pro-thrombotic state in adult life. An altered inflammatory state could be one link between small newborn/infant size and adult cardiovascular disease. Associations between pro-inflammatory markers and childhood/adolescent BMI gain are probably mediated through adult adiposity.
Birthweight; growth; C-reactive protein; fibrinogen; plasminogen activator inhibitor-1
Neurotrophin 3 (Ntf3) is expressed in Sertoli cells and acts as a chemo-attractant for cell migration from the mesonephros into the developing testis, a process critical to the early morphological events of testis cord formation. The male sex-determining gene Sry initiates the process of testicular development. Sox9 is a key regulator of male sex determination and is directly regulated by SRY. Information on other downstream target genes of SRY is limited. The current study demonstrates an interaction of SRY with the Ntf3 promoter both in vitro and in vivo. The Ntf3 promoter in both rat and mouse contains at least one putative SRY binding site in the −0.6 kb promoter region. In a luciferase reporter assay system, both SRY and SOX9 stimulated the Ntf3 promoter in vitro through an interaction with this SRY-binding motif. In an immunoprecipitation-based pull-down assay, recombinant SRY protein bound the Ntf3 promoter fragment containing an intact SRY binding site, whereas the same protein did not interact with the fragment containing a mutated SRY motif. Specific antibodies against SRY were used in a chromatin immunoprecipitation (ChIP) assay of embryonic testis and were found to precipitate the Ntf3 promoter region. The SRY ChIP assay confirmed the direct interaction between SRY and the Ntf3 promoter in vivo during male sex determination. Observations suggest that SRY physically interacts with the Ntf3 promoter during male sex determination to coordinate cell migration in the testis to form testis cords.
SRY directly regulates the Ntf3 promoter in vitro and in vivo during male sex determination.
Ntf3; rat; sex determination; Sry; testis development
The risk of type 2 diabetes mellitus is increased in people who have low birth weights and who subsequently become obese as adults. Whether their obesity originates in childhood and, if so, at what age are unknown. Understanding the origin of obesity may be especially important in developing countries, where type 2 diabetes is rapidly increasing yet public health messages still focus on reducing childhood “undernutrition.”
We evaluated glucose tolerance and plasma insulin concentrations in 1492 men and women 26 to 32 years of age who had been measured at birth and at intervals of three to six months throughout infancy, childhood, and adolescence in a prospective, population-based study.
The prevalence of impaired glucose tolerance was 10.8 percent, and that of diabetes was 4.4 percent. Subjects with impaired glucose tolerance or diabetes typically had a low body-mass index up to the age of two years, followed by an early adiposity rebound (the age after infancy when body mass starts to rise) and an accelerated increase in body-mass index until adulthood. However, despite an increase in body-mass index between the ages of 2 and 12 years, none of these subjects were obese at the age of 12 years. The odds ratio for disease associated with an increase in the body-mass index of 1 SD from 2 to 12 years of age was 1.36 (95 percent confidence interval, 1.18 to 1.57; P<0.001).
There is an association between thinness in infancy and the presence of impaired glucose tolerance or diabetes in young adulthood. Crossing into higher categories of body-mass index after the age of two years is also associated with these disorders.
cardiovascular risk factors; cohort; incidence; Indian Asians
To report the successful outcome of a rare optic pit-associated maculopathy with an outer retinal hole following 23 G vitrectomy, internal limiting membrane (ILM) peeling and fluid-gas exchange without additional endolaser.
Interventional case report.
This case report documents a 56-year-old male patient with complaints of progressive diminution of vision in the right eye more than in the left eye due to an optic disc pit with an outer retinal hole and a cataract. Optical coherence tomography confirmed the presence of an outer retinal hole. The case report shows the successful outcome of a rare optic pit-associated maculopathy with an outer retinal hole and a cataract following phacoemulsification with 23 G vitrectomy, ILM peeling and fluid-gas exchange without additional endolaser.
Optic pit; Outer retinal hole; Retinoschisis; Vitrectomy
To study the relationship between lipid peroxidation of spermatozoa and changes in functional intergrity of human spermatozoa membrane in male subjects.
Materials and Methods
Sixty eight male partners of infertile couples were included in the study. Status of oxidative stress was assessed by determining malondialdehyde (MDA) in seminal plasma. Functinal intergrity of sperm membrance was tested subjecting the sperm to hypo-osmotic test (HOS). The seminal plasma MDA levels were compared with seminogram parameters as well as with the results of HOS test using Pearson’s coefficient of correlation (r) and significance of coefficient of correlation calculated from the table.
A significant but weak negative correlation was observed between seminal plasma MDA level and sperm concentration (r=−0.33,p<0.05), sperm motility (r=−0.37,p<0.05), sperm morphology (r=−0.37,p<0.05), and percentage of HOS positive spermatozoa (r=−0.33,p<0.05). Percentages of HOS positive spermatozoa also exhibited a significant but weak negative relationship with sperm concentration (r=−0.47,p<0.01), sperm motility (r=−0.48,p<0.01), sperm morphology (r=−0.49,p<0.01).
Lipid peroxidation of spermatozoa is likely to affect the functional intergrity of the spermatozoa membrane.
oxidative stress; spermatozoa; membrane integrity
The present study has been undertaken to apply the concept of nanoparticulate mucopenetrating drug delivery system for complete eradication of Helicobacter pylori (H. pylori), colonised deep into the gastric mucosal lining. Most of the existing drug delivery systems have failed on account of either improper mucoadhesion or mucopenetration and no dosage form with dual activity of adhesion and penetration has been designed till date for treating H. pylori induced disorders. In the present study, novel chitosan-alginate polyelectrolyte complex (CS-ALG PEC) nanoparticles of amoxicillin have been designed and optimized for various variables such as pH and mixing ratio of polymers, concentrations of polymers, drug and surfactant, using 33 Box-Behnken design. Various studies like particle size, surface charge, percent drug entrapment, in-vitro mucoadhesion and in-vivo mucopenetration of nanoparticles on rat models were conducted. The optimised FITC labelled CS-ALG PEC nanoparticles have shown comparative low in-vitro mucoadhesion with respect to plain chitosan nanoparticles, but excellent mucopenetration and localization as observed with increased fluorescence in gastric mucosa continuously over 6 hours, which clinically can help in eradication of H. pylori.
Factorial design; Mucopenetration; Helicobacter pylori; Box Behnken design; Stomach specific delivery system