Spectrum of illness among returned Australian travellers from Bali, Indonesia: a 5-year retrospective observational study.

BACKGROUND: Bali, Indonesia, presents significant infectious and non-infectious health risks for Australian travellers. Understanding this spectrum of illnesses has the potential to assist clinicians in evaluating unwell returning travellers and guide provision of pre-travel advice. AIM: To describe the spectrum of illnesses seen in returned travellers from Bali. METHODS: Using a novel text mining approach, we performed a retrospective, observational study of all adult emergency department presentations to a metropolitan health service in Melbourne, from 2011 to 2015. Outcome measures included demographic, clinical and laboratory features of travel-related illnesses. RESULTS: A total of 464 patients met inclusion criteria. Gastroenteritis (119/464, 26%), systemic febrile illness (88/464, 19%) and respiratory tract infection (51/464, 11%) were the most common diagnoses. Dengue was the most common laboratory-confirmed diagnosis (25/464, 5%). No cases of malaria were identified. Common non-infectious presentations included traumatic injury (47/464, 10%) and animal bites requiring rabies post-exposure prophylaxis (29/464, 6%). A total of 110 patients (24%) was admitted to the hospital; those presenting with systemic febrile illness were more likely to be admitted compared to those presenting with other illnesses (odds ratio 3.42, 95% confidence interval 2.02-5.75, P < 0.001). CONCLUSION: This is the first study to use a text mining approach to identify and describe emergency department presentations related to diseases acquired in Bali by Australian travellers. Although infections are important causes of illness, trauma and animal bites account for a significant number of hospital presentations. Our findings contribute to the knowledge on the health risks for travellers to Bali, and will assist clinicians in relevant pre- and post-travel evaluations.

Cryptococcal infection in lung transplant recipients: A 5-year retrospective review at an Australian transplant center.

Cryptococcosis is a common invasive fungal infection (IFI) in solid organ transplant (SOT) recipients. Little is known about cryptococcosis in lung transplant (LTx) recipients despite having one of the highest risks of infection. The aim of this study was to describe demographic and clinical features of cryptococcal infection in LTx recipients. We performed a retrospective, observational study of cryptococcal infection in LTx recipients at The Alfred Hospital in Melbourne, Australia, from 2012 to 2017. A total of 11 cases were identified. Seven patients (64%) were male and the median age was 54.7 years (range 34-69 years). Diagnosis occurred at a median of 233 days (range 1-3650 days) post-transplant. Nine patients (82%) had isolated pulmonary infection of whom 7 (78%) were asymptomatic. All were treated with oral antifungal therapy and 1 required surgical resection of infected lung. Two patients (18%) had disseminated infection; 1 with pulmonary and central nervous system (CNS) infection and 1 with isolated CNS infection. Both patients presented with headache and brain imaging demonstrated cerebral edema, myelinosis, and leptomeningeal enhancement. One of these patients died. This study highlights the fact that cryptococcal infection should remain a consideration in asymptomatic LTx recipients, especially in the presence of non-specific nodules on chest imaging, and that the presence of headache in these patients requires urgent investigation for CNS infection.

Donor-Derived Mycoplasma hominis and an Apparent Cluster of M. hominis Cases in Solid Organ Transplant Recipients.

BACKGROUND: Invasive and disseminated Mycoplasma hominis infections are well recognized but uncommon complications in solid organ transplant recipients. In a single center, a cluster of M. hominis infections were identified in lung transplant recipients from the same thoracic intensive care unit (ICU). We sought to determine the source(s) of these infections. METHODS: Medical records of the donor and infected transplant recipients were reviewed for clinical characteristics. Clinical specimens underwent routine processing with subculture on Mycoplasma-specific Hayflick agar. Mycoplasma hominis identification was confirmed using sequencing of the 16S ribosomal RNA gene. Mycoplasma hominis isolates were subjected to whole-genome sequencing on the Illumina NextSeq platform. RESULTS: Three lung transplant recipients presented with invasive M. hominis infections at multiple sites characterized by purulent infections without organisms detected by Gram staining. Each patient had a separate donor; however, pretransplant bronchoalveolar lavage fluid was only available from the donor for patient 1, which subsequently grew M. hominis. Phylo- and pangenomic analyses indicated that the isolates from the donor and the corresponding recipient (patient 1) were closely related and formed a distinct single clade. In contrast, isolates from patients 2 and 3 were unrelated and divergent from one another. CONCLUSIONS: Mycoplasma hominis should be considered a cause of donor-derived infection. Genomic data suggest donor-to-recipient transmission of M. hominis. Additional patients co-located in the ICU were found to have genetically unrelated M. hominis isolates, excluding patient-to-patient transmission.

Increase in Meningococcal Serogroup W Disease, Victoria, Australia, 2013-2015.

In Victoria, Australia, invasive meningococcal disease caused by Neisseria meningitidis serogroup W increased from 4% of all cases in 2013 to 30% in 2015. This increase resulted largely from strains similar to those in the serogroup W sequence type 11 clonal complex, previously described in the United Kingdom and South America.

Lactobacillus acidophilus: characterization of the species and application in food production.

L. acidophilus is a homofermentative, microaerophilic, short chain gram positive microorganism with rod morphology having its bacteriocins belonging to class II a. Several bacteriocins of L. acidophilus have been isolated and characterized. These are structurally similar, but their molecular weight varies as well as their spectrum of antimicrobial activity. They exhibit important technical properties, i.e., thermostability and retaining of activity at a wide pH range along with strong inhibitory actions against food spoilage and pathogenic bacteria make them an important class of biopreservatives. L. acidophilus can be added as an adjunct in many food fermentation processes contributing to unique taste, flavor, and texture. It also preserves the products by producing lactic acid and bacteriocins. A lot of new information regarding the bacteriocins of L. acidophilus has emerged during the last few years. In this review, an attempt has been made to summarize and discuss all the available information regarding the sources of bacteriocins production, their characteristics, and their antimicrobial action along with their application.

Some probiotic and antibacterial properties of Lactobacillus acidophilus cultured from dahi a native milk product.

In this study, different strains of Lactobacillus acidophilus from dahi were analyzed for certain probiotic and antibacterial properties. Initially, these strains were confirmed by the amplification of 16S rRNA regions and then screened for antibacterial activities against food borne pathogens. The phenotypic relationship between apparent antibacterial activity and cell wall proteins were established by cluster analysis. It was observed that those strains, which have prominent bands having size 22-25 kDa possess antibacterial activity. On the basis of wide spectrum of killing pattern, a strain LA06FT was further characterized that showed no change in its behavior when subjected to the antibiotic protected environment and grow well in acid-bile conditions. The bacteriocin produced by this strain has specific antibacterial activity of 5369.13 AU mg(-1). It remained stable at 60-90 °C and pH range of 4.5-6.5 while proteolytic enzymes inactivate the bacteriocin that confirm its proteinic nature having molecular weight of ≤8.5 kDa.

Dengue Fever in Immunocompromised Patients: A Systematic Review and Meta-Analysis.

OBJECTIVE: Given the complex role of immunity in dengue severity, we aimed to review the clinical course of dengue infection in immunocompromised patients. METHODS: We conducted a systematic review of studies reporting outcomes among immunocompromised patients with laboratory-confirmed dengue infection. Meta-analysis using the Mantel-Haenszel method (fixed effects) was performed for studies with control groups. We registered the study with PROSPERO (No. CRD42021258930). RESULTS: We included 115 studies. Among these, 30 studies compared immunocompromised (cases) and non-immunocompromised (control) patients, focusing mainly on children (n=22 studies) with malnutrition (n=18). Immunocompromised patients had a higher likelihood of dengue complications (OR 1.87; 95% CI: 1.04 - 3.35]) but a lower likelihood of severe dengue (OR 0.83; 95% CI: 0.69 - 1.00]. No significant difference in mortality was observed. In the 85 studies focused solely on immunocompromised patients, severe dengue and mortality rates were 9% and 4%, respectively, mostly among adult solid organ transplant recipients and those with inflammatory diseases. Immunosuppressive treatment alterations and temporary graft dysfunction were reported. CONCLUSIONS: Immunocompromised patients have an increased risk of dengue-related complications. However, definitive conclusions about the comparative severity of dengue across different immunocompromised patient groups are limited by a lack of robust data, highlighting the need for well-designed future studies.

The epidemiology of imported and locally acquired dengue in Australia, 2012-2022.

BACKGROUND: Dengue is the most important arboviral disease globally and poses ongoing challenges for control including in non-endemic countries with competent mosquito vectors at risk of local transmission through imported cases. We examined recent epidemiological trends in imported and locally acquired dengue in Australia, where the Wolbachia mosquito population replacement method was implemented throughout dengue-prone areas of northern Queensland between 2011 and 2019. METHODS: We analysed dengue cases reported to the Australian National Notifiable Disease Surveillance System between January 2012 and December 2022, and Australian traveller movement data. RESULTS: Between 2012 and 2022, 13 343 dengue cases were reported in Australia (median 1466 annual cases); 12 568 cases (94.2%) were imported, 584 (4.4%) were locally acquired and 191 (1.4%) had no origin recorded. Locally acquired cases decreased from a peak in 2013 (n = 236) to zero in 2021-22. Annual incidence of imported dengue ranged from 8.29/100 000 (n = 917 cases) to 22.10/100 000 (n = 2203) annual traveller movements between 2012 and 2019, decreased in 2020 (6.74/100 000 traveller movements; n = 191) and 2021 (3.32/100 000 traveller movements; n = 10) during COVID-19-related border closures, then rose to 34.79/100 000 traveller movements (n = 504) in 2022. Imported cases were primarily acquired in Southeast Asia (n = 9323; 74%), Southern and Central Asia (n = 1555; 12%) and Oceania (n = 1341; 11%). Indonesia (n = 5778; 46%) and Thailand (n = 1483; 12%) were top acquisition countries. DENV-2 (n = 2147; 42%) and DENV-1 (n = 1526; 30%) were predominant serotypes. CONCLUSION: Our analysis highlights Australia's successful control of locally acquired dengue with Wolbachia. Imported dengue trends reflect both Australian travel destinations and patterns and local epidemiology in endemic countries.

Imported malaria into Australia: surveillance insights and opportunities.

BACKGROUND: Malaria continues to pose a significant burden in endemic countries, many of which lack access to molecular surveillance. Insights from malaria cases in travellers returning to non-endemic areas can provide valuable data to inform endemic country programmes. To evaluate the potential for novel global insights into malaria, we examined epidemiological and molecular data from imported malaria cases to Australia. METHODS: We analysed malaria cases reported in Australia from 2012 to 2022 using National Notifiable Disease Surveillance System data. Molecular data on imported malaria cases were obtained from literature searches. RESULTS: Between 2012 and 2022, 3204 malaria cases were reported in Australia. Most cases (69%) were male and 44% occurred in young adults aged 20-39 years. Incidence rates initially declined between 2012 and 2015, then increased until 2019. During 2012-2019, the incidence in travellers ranged from 1.34 to 7.71 per 100 000 trips. Cases were primarily acquired in Sub-Saharan Africa (n = 1433; 45%), Oceania (n = 569; 18%) and Southern and Central Asia (n = 367; 12%). The most common countries of acquisition were Papua New Guinea (n = 474) and India (n = 277). Plasmodium falciparum accounted for 58% (1871/3204) of cases and was predominantly acquired in Sub-Saharan Africa, and Plasmodium vivax accounted for 32% (1016/3204), predominantly from Oceania and Asia. Molecular studies of imported malaria cases to Australia identified genetic mutations and deletions associated with drug resistance and false-negative rapid diagnostic test results, and led to the establishment of reference genomes for P. vivax and Plasmodium malariae. CONCLUSIONS: Our analysis highlights the continuing burden of imported malaria into Australia. Molecular studies have offered valuable insights into drug resistance and diagnostic limitations, and established reference genomes. Integrating molecular data into national surveillance systems could provide important infectious disease intelligence to optimize treatment guidelines for returning travellers and support endemic country surveillance programmes.

Perceptions of travel-related health risks and pre-travel health-seeking behaviour among students: A qualitative analysis.

BACKGROUND: The impact of the COVID-19 pandemic on travel risk perceptions and pre-travel healthcare attendance is unknown. We aimed to explore barriers and enablers to engagement in and uptake of pre-travel advice (PTA) among university students, a recognised at-risk group for travel-related morbidity and poor PTA uptake. Additionally, we aimed to explore whether the COVID-19 pandemic has influenced attitudes towards future PTA. METHODS: Semi-structured interviews were conducted with university students (October-December 2021). Thematic analysis of the data was conducted using the Health Belief Model (HBM) framework. RESULTS: We interviewed 18 students (11 male, median age 24). Students' risk perceptions relating to travel-related illnesses were influenced by past experiences, travel destination, vaccination status and knowledge. Barriers to seeking PTA included a lack of perceived need and concerns about accessibility/availability, whereas potential benefits included reduced health risks, vaccine-induced protection, and knowledge gain. Cues to seeking future PTA were perceived risks of travel-related illness, perceived high-risk destination, itinerary-related vaccine requirements, perceived benefits of advice and concerns about COVID-19 infection. COVID-19 risk perceptions did not consistently align with those of other travel-related illness. We used findings to inform recommendations. CONCLUSION: The current COVID-19 pandemic offers an opportunity to increase students' risk awareness and promote preventive travel health behaviours. Recommendations to improve students' future engagement in PTA include: 1) increasing awareness of risks associated with travel, 2) emphasising PTA benefits, and 3) ensuring PTA is convenient, accessible and affordable.

Current roles of lignin for the agroindustry: Applications, challenges, and opportunities.

Lignin has the potential to be used as an additive, coating agent, fertilizer, plant growth stimulator, and packaging material in the agroindustry due to its functional aromatic structure. The quantitative measurement of functional groups is a significant element of the research for lignin structure since they directly impact their optical, dispersion, and chemical properties. These physical and chemical properties of lignin strongly depend on its type and source and its isolation procedure. Thus, lignin provides numerous opportunities for the circular economy in the agroindustry; however, studying and resolving the challenges associated with its separation, purification, and modification is required. This review discusses the most recent findings on lignin use in agroindustry and historical facts about lignin. The properties of lignin and its roles as coating agents, pesticide carriers, plant growth stimulators, and soil-improving agents have been summarized. The emerging challenges in the field of lignin-based agroindustry are considered, and potential future steps to overcome these challenges are discussed.

Recent Innovations in Non-dairy Prebiotics and Probiotics: Physiological Potential, Applications, and Characterization.

Non-dairy sources of prebiotics and probiotics impart various physiological functions in the prevention and management of chronic metabolic disorders, therefore nutraceuticals emerged as a potential industry. Extraction of prebiotics from non-dairy sources is economical and easily implemented. Waste products during food processing, including fruit peels and fruit skins, can be utilized as a promising source of prebiotics and considered "Generally Recognized As Safe" for human consumption. Prebiotics from non-dairy sources have a significant impact on gut microbiota and reduce the population of pathogenic bacteria. Similarly, next-generation probiotics could also be isolated from non-dairy sources. These sources have considerable potential and can give novel strains of probiotics, which can be the replacement for dairy sources. Such strains isolated from non-dairy sources have good probiotic properties and can be used as therapeutic. This review will elaborate on the potential non-dairy sources of prebiotics and probiotics, their characterization, and significant physiological potential.

In vitro screening of EPS-producing Streptococcus thermophilus strains for their probiotic potential from Dahi.

Dahi is a very common and traditional fermented dairy product in Pakistan and its neighboring countries, it represents a rich source for the isolation of many new strains of lactic acid bacteria (LAB). The major objective of this study was to evaluate the probiotic potential of novel exopolysaccharide (EPS)-producing strains of S. thermophilus isolated from Dahi, sold in the local markets of Rawalpindi and Islamabad, Pakistan. In this study, 32 isolates of S. thermophilus were initially isolated from Dahi and out of these, 10 identified strains were further screened for their EPS-producing ability. Maximum EPS production was estimated for RIY strain (133.0 ± 0.06), followed by RIH4 strain (103.83 ± 0.76) and RIRT2 strain (95.77 ± 0.22), respectively. Thereafter, in vitro studies revealed that these newly identified EPS-producing strains of S. thermophilus fulfilled the basic requirements for probiotic functions; including resistance to harsh conditions of GIT, good cell surface hydrophobicity, auto-aggregation, and co-aggregation, especially against L. monocytogenes. Finally, the safety assessment displayed that these strains were also sensitive to clinical antibiotics, including vancomycin. Thus, these selected EPS strains of S. thermophilus act as potential candidates for biostabilizers in the preparation of consumer-friendly fermented probiotic milk products.

Gastrointestinal transit tolerance, cell surface hydrophobicity, and functional attributes of Lactobacillus Acidophilus strains isolated from Indigenous Dahi.

Strains of Lactobacillus acidophilus WFA1 (KU877440), WFA2 (KU877441), and WFA3 (KU877442) were isolated from indigenous Dahi (yogurt), screened, and selected based on acid and bile tolerance along with the antimicrobial activity. These selected strains were further assessed for their probiotic and functional attributes. Results for simulated gastric and intestinal tolerance/ resistance revealed that all three strains can resist and survive under the following mentioned conditions. To access cell surface hydrophobicity, bacterial adhesion to hydrocarbons (BATH), cellular auto-aggregation, and salt aggregation were performed. In BATH, adhesion of strains against three hydrocarbons namely xylene, dichloromethane, and hexadecane was conducted. The results show that strains showed the least adhesion to xylene (54.25%) as compared to dichloromethane (55.25%) and hexadecane (56.65%). WFA1 showed maximum adherence percentage (55.48%) followed WFA2 (55.48%) and WFA3 (51.38%). Cellular auto-aggregation varied from 21.72% to 30.73% for WFA3 and WFA1, respectively. In the salt aggregation test (SAT), WFA1, WFA2, and WFA3 aggregated at 0.6, 1.0, and 2.0 molar concentrations of ammonium sulfate, respectively. PCR amplification of bile salt hydrolase gene (bsh) was performed and sequences were submitted to the public database of NCBI and Gene bank under accession numbers, KY689139, KY689140, and KY689141. Additionally, a cholesterol-lowering assay was conducted and up to 26% reduction in cholesterol was observed by the strains. Regarding functional properties, exopolysaccharide (EPS) production, and antioxidant potential, strain WFA1 showed promising results EPS (1.027mg/ml), DPPH (80.66%), ABTS (81.97%), and reducing power (1.787). It can be concluded from the present study that the mentioned strains of L. acidophilus (WFA1, WFA2, and WFA3) are strongly hydrophobic; thus having an ability to survive and colonize under the gastrointestinal tract which confirms their probiotic nature. Regarding their functional properties, L. acidophilus WFA1 (KU877440) showed excellent properties of antioxidants and EPS production.

An unexpected diagnosis of human immunodeficiency virus-2 infection in an overseas visitor: a case report.

BACKGROUND: Human immunodeficiency virus 2 infection is endemic in West Africa but is also found in parts of Europe, North and South America, and India where it is thought to have been introduced secondary to migration and commercial trade ties. It is less common than Human immunodeficiency virus 1, with differences in pathogenicity, lower rates of transmission, longer asymptomatic period and slower progression to acquired immunodeficiency syndrome. Human immunodeficiency virus 2 is also associated with diagnostic challenges given the lack of commercially available diagnostic tests, and management challenges given intrinsic resistance to many anti-retroviral therapies. CASE PRESENTATION: We describe a case of a 65 year old South Indian female, visiting her family in Australia, who presented with weight loss, pancytopaenia and generalised lymphadenopathy on a background of newly diagnosed congestive cardiac failure. Multiple investigations were performed to elucidate the cause of her presentation, with the eventual unexpected diagnosis of human immunodeficiency virus 2. She was commenced on anti-retroviral treatment and made a remarkable recovery. CONCLUSION: We describe the challenges associated with diagnosis of human immunodeficiency virus 2 due to lack of commercially available diagnostics, as well as the treatment and management challenges including the fact that human immunodeficiency virus 2 is intrinsically resistant to non-nucleoside reverse transcriptase inhibitors. Human immunodeficiency virus 2 infection should be considered in patients who present with symptoms and signs that do not point towards a clear diagnosis, such as unexplained pancytopaenia or lymphadenopathy, and who have risk factors such as being from an endemic area or having had blood transfusions, especially prior to the commencement of blood-borne virus screening of blood donors.

Travel vaccination recommendations and endemic infection risks in solid organ transplantation recipients.

BACKGROUND: Solid organ transplant (SOT) recipients are often heavily immunosuppressed and consequently at risk of serious illness from vaccine preventable viral and bacterial infections or with endemic fungal and parasitic infections. We review the literature to provide guidance regarding the timing and appropriateness of vaccination and pathogen avoidance related to the immunological status of SOT recipients. METHODS: A PUBMED search ([Vaccination OR vaccine] AND/OR ["specific vaccine"] AND/OR [immunology OR immune response OR cytokine OR T lymphocyte] AND transplant was performed. A review of the literature was performed in order to develop recommendations on vaccination for SOT recipients travelling to high-risk destinations. RESULTS: Whilst immunological failure of vaccination in SOT is primarily the result of impaired B-cell responses, the role of T-cells in vaccine failure and success remains unknown. Vaccination should be initiated at least 4 weeks prior to SOT or more than 6 months post-SOT. Avoidance of live vaccination is generally recommended, although some live vaccines may be considered in the specific situations (e.g. yellow fever). The practicing physician requires a detailed understanding of region-specific endemic pathogen risks. CONCLUSIONS: We provide a vaccination and endemic pathogen guide for physicians and travel clinics involved in the care of SOT recipients. In addition, recommendations based on timing of anticipated immunological recovery and available evidence regarding vaccine immunogenicity in SOT recipients are provided to help guide pre-travel consultations.

Effects of glutathione-S-transferase polymorphisms on the risk of breast cancer: a population-based case-control study in Pakistan.

Cancer is widely accepted as one of the major health issues. Diet composition and exposure to environmental genotoxic and carcinogenic agents such as polycyclic aromatic hydrocarbons (PAHs) are among the causative factors for various types of cancers, including breast cancer. Low penetrance genes including glutathione S transferases (GST) in association with environmental factors can contribute greatly in the development of breast cancer. We were interested to investigate the association of the polymorphisms of GSTM1, GSTT1, GSTP1 and GSTO2 with the risk of breast cancer in the Pakistani population. One hundred women visiting the Department of Radiology and Oncology, Nishter Hospital, Multan with pathologically confirmed breast cancer, and 100 healthy volunteers from central Pakistan were enrolled in the present study. The strength of the association of various factors with breast cancer was measured by calculating odd ratios (ORs) which were determined by logistic regression. All P values cited are two-sided; differences resulting in a P value of less or equal to 0.05 were declared statistically significant. The Hardy Weinberg equilibrium was tested for the genotype proportions in the control group, as a measure of quality control. Those aged 36-45, in menopause or with a history of cancer in the family had a significantly higher prevalence of breast cancer compared with controls. The frequency of GSTM1 and GSTT1 was similar in both control and patients suggesting no association with the risk of cancer development, however GSTM1 and GSTT1 were significantly linked with the risk of breast cancer in smokers and in women with a history of breast cancer in the family respectively. Similarly women homozygous for GSTP1 or GSTO2 and with a history of breast cancer, or in menopause, were at greater risk of breast cancer than wild type or heterozygotes. Our data suggest that genetic differences in some GST genes may be linked with an increased susceptibility to breast cancer. Furthermore it also gives an insight into the interaction between the GST polymorphisms and pre-menopausal diagnosis of breast cancer.

Evaluation of Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM encapsulated using a novel impinging aerosol method in fruit food products.

This study investigated the effect of microencapsulation on the survival of Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM and their acidification in orange juice at 25°C for nine days and at 4°C over thirty five days of storage. Alginate micro beads (10-40 µm) containing the probiotics were produced by a novel dual aerosol method of alginate and CaCl(2) cross linking solution. Unencapsulated L. rhamnosus GG was found to have excellent survivability in orange juice at both temperatures. However unencapsulated L. acidophilus NCFM showed significant reduction in viability. Encapsulation of these two bacteria did not significantly enhance survivability but did reduce acidification at 25°C and 4°C. In agreement with this, encapsulation of L. rhamnosus GG also reduced acidification in pear and peach fruit-based foods at 25°C, however at 4°C difference in pH was insignificant between free and encapsulated cells. In conclusion, L. rhamnosus GG showed excellent survival in orange juice and microencapsulation has potential in reducing acidification and possible negative sensory effects of probiotics in orange juice and other fruit-based products.

Molecular genetic studies and delineation of the oculocutaneous albinism phenotype in the Pakistani population.

BACKGROUND: Oculocutaneous albinism (OCA) is caused by a group of genetically heterogeneous inherited defects that result in the loss of pigmentation in the eyes, skin and hair. Mutations in the TYR, OCA2, TYRP1 and SLC45A2 genes have been shown to cause isolated OCA. No comprehensive analysis has been conducted to study the spectrum of OCA alleles prevailing in Pakistani albino populations. METHODS: We enrolled 40 large Pakistani families and screened them for OCA genes and a candidate gene, SLC24A5. Protein function effects were evaluated using in silico prediction algorithms and ex vivo studies in human melanocytes. The effects of splice-site mutations were determined using an exon-trapping assay. RESULTS: Screening of the TYR gene revealed four known (p.Arg299His, p.Pro406Leu, p.Gly419Arg, p.Arg278*) and three novel mutations (p.Pro21Leu, p.Cys35Arg, p.Tyr411His) in ten families. Ex vivo studies revealed the retention of an EGFP-tagged mutant (p.Pro21Leu, p.Cys35Arg or p.Tyr411His) tyrosinase in the endoplasmic reticulum (ER) at 37°C, but a significant fraction of p.Cys35Arg and p.Tyr411His left the ER in cells grown at a permissive temperature (31°C). Three novel (p.Asp486Tyr, p.Leu527Arg, c.1045-15 T > G) and two known mutations (p.Pro743Leu, p.Ala787Thr) of OCA2 were found in fourteen families. Exon-trapping assays with a construct containing a novel c.1045-15 T > G mutation revealed an error in splicing. No mutation in TYRP1, SLC45A2, and SLC24A5 was found in the remaining 16 families. Clinical evaluation of the families segregating either TYR or OCA2 mutations showed nystagmus, photophobia, and loss of pigmentation in the skin or hair follicles. Most of the affected individuals had grayish-blue colored eyes. CONCLUSIONS: Our results show that ten and fourteen families harbored mutations in the TYR and OCA2 genes, respectively. Our findings, along with the results of previous studies, indicate that the p.Cys35Arg, p.Arg278* and p.Gly419Arg alleles of TYR and the p.Asp486Tyr and c.1045-15 T > G alleles of OCA2 are the most common causes of OCA in Pakistani families. To the best of our knowledge, this study represents the first documentation of OCA2 alleles in the Pakistani population. A significant proportion of our cohort did not have mutations in known OCA genes. Overall, our study contributes to the development of genetic testing protocols and genetic counseling for OCA in Pakistani families.

Survivability of probiotics encapsulated in alginate gel microbeads using a novel impinging aerosols method.

Encapsulation of probiotic bacteria in cross-linked alginate beads is of major interest for improving the survivability in harsh acid and bile environment and also in food matrices. Alginate micro beads (10-40 µm) containing the probiotics Lactobacillus rhamnosus GG and Lactobacillus acidophilus NCFM were produced by a novel technique based on dual aerosols of alginate solution and CaCl(2) cross linking solution. Extruded macro beads (approximately 2mm diameter) produced by the conventional method and micro beads produced by novel aerosols technique offered comparable protection to L. rhamnosus in high acid and bile environment. Chitosan coating of micro beads resulted in a significant increase in survival time of L. rhamnosus from 40 to 120 min in acid condition and the reduction in cell numbers was confined to 0.94 log over this time. Alginate macro beads are more effective than micro beads in protecting L. acidophilus against high acid and bile. Chitosan coating of micro beads resulted in similar protection to L. acidophilus in macro beads in acid and extended the survival time from 90 to at least 120 min. Viability of this organism in micro beads was 3.5 log after 120 min. The continuous processing capability and scale-up potential of the dual aerosol technique offers potential for an efficient encapsulation of probiotics in very small alginate micro beads below sensorial detection limits while still being able to confer effective protection in acid and bile environment.