Improved Pregnancy Outcomes and Endometrial Receptivity by Thawed Frozen Embryo Transfer in Mildly Stimulated Cycles with Letrozole Combined with Estrogen in Women with Unresponsive Thin Endometrium Compared to Standard Endometrial Preparation with Estrogen Alone: A Retrospective Study.

Context: Infertile women undergoing frozen embryo transfer (FET) cycles may not show optimal endometrial growth with estrogens alone. Aim: To evaluate clinical effect of mild stimulation with letrozole and estrogens on endometrial growth in comparison to standard endometrial preparation with oral and topical estrogens in infertile women with unresponsive thin endometrium undergoing FET. Settings and design: Retrospective observational case-control study. Material and methods: Forty women unresponsive to first AC-FET cycle were given mild stimulation with letrozole and estrogens as second LE-FET cycle for endometrial preparation (LE-FET study group) and compared with 40 historical controls who had received two cycles of AC-FET (AC-FET control group). Responses were assessed by optimal endometrial thickness (≥ 7 mm) and clinical pregnancy. Statistical analysis: Descriptive statistics were elaborated by mean ± SD and percentages. Results were expressed by mean ± SD, unpaired t test for difference in endometrial thickness, chi square and Fisher exact test to compare the difference in pregnancy among both groups. Results: Mean endometrial thickness was significantly increased in LE-FET study group (6.68 ± 2.09 mm) versus AC-FET control group (5.35 ± 1.90 mm). Higher clinical pregnancy rate was noted in study group as compared to control group (35% versus 12.5%). Conclusion: This study suggests that letrozole with estradiol (LE-FET) compared to estradiol alone (AC-FET) for second cycle significantly increased endometrial thickness and improved clinical pregnancy rates in women with unresponsive thin endometrium after first AC-FET cycle with estradiol alone. Addition of letrozole to estrogen upfront for FET cycles may enhance endometrial receptivity and might improve pregnancy outcomes.

Changing epidemiology of SARS-CoV in the context of COVID-19 pandemic.

SARS-CoV-2 is a new form of ß-coronavirus that has been recently discovered and is responsible for COVID 19 pandemic. The earliest infection can be traced back to Wuhan, China. From there it has spread all over the world. Keeping in view the above perspective, an attempt is made in order to find out the epidemiological pattern of COVID 19 pandemic, if any, in different geo-climatological regions of the world in terms of case incidence and mortality. This study is also an endeavor to review and analyze the gradual changes of the genetic makeup of SARS-CoV from evolutionary and epidemiological perspectives. The raw data of COVID-19 cases and death incidences were collected from the World Health Organization (WHO) website from the time period: 1st April to 6th April, 2020. The data that are utilized here for general and Case fatality rate (CFR) based analysis. Western pacific region, European region and Americas have the greatest number of infected cases (P < 0.001); whereas deaths have been found to be significantly higher in Europe (P < 0.001). Total number of confirmed cases and deaths in south-east Asia are comparatively lower (P < 0.001). Case fatality rate (CFR) has also found significant for European region. SARS-CoV-2 is considered to be a strain of SARS-CoV that has a high rate of pathogenicity and transmissibility. Result indicated that the European region has been affected mostly for both cases and death incidences. The novel mutations in SARS-CoV-2 possibly increase the virus infectivity. Genetic heterogeneity of this virus within the human population might originate as the representatives of naturally selected virus quasispecies. In this context, the presence of the asymptomatic individuals could be a significant concern for SARS-CoV-2 epidemiology. Further studies are required to understand its genetic evolution and epidemiological significance.

PRESENCE OF BOTH DENGUE SPECIFIC NS1 ANTIGEN AND IMMUNOGLOBULIN M ANTIBODY IN CONFIRMED DENGUE PATIENTS IN KOLKATA

@#The object of this study was to identify patients with diagnosed dengue infection, who were positive for both dengue-specific NS1 antigen and IgM antibody.

Tyrosinase inhibitor-loaded microsponge drug delivery system: new approach for hyperpigmentation disorders.

OBJECTIVE: The aim of the present investigation was to design and formulate appropriate form of glabridin, using microsponge drug delivery system. METHOD: Microsponges were prepared by emulsion solvent evaporation method and characterized by drug loading, infrared spectroscopy and scanning electron microscopy. In vitro diffusion studies of gel formulation were performed using ethanol: phosphate buffer (1:1) mixture as receptor medium. Animal studies were carried out using brownish guinea pigs with UV-induced pigmentation model. RESULTS: Prepared microsponges were predominantly yellowish, free-flowing and spherical in shape. The infrared spectra revealed the absence of drug polymer interaction. Scanning electron microscopy (SEM) and porosity studies confirmed spherical and porous nature. In vitro release studies data depicted highest correlation with Higuchi treatment. Animal studies also supported the better depigmenting activity as compared to plain gel. CONCLUSION: Glabridin microsponge-loaded gel could be efficacious in treating various hyperpigmentation disorders.

Designing of the mucoadhesive intravaginal spermicidal films.

A mucoadhesive drug delivery system for local delivery of metronidazole through vaginal route was formulated. Films were prepared by solvent evaporation method using various compositions of Carbopol, hydroxypropylmethylcellulose, chitosan, Polyox and propylene glycol. The films were evaluated for their weight, thickness, surface pH, folding endurance, mechanical, drug content uniformity, in vitro drug release, swelling, gelling and mucoadhesion property. All the films possess satisfactory film characteristics. Films made of Polyox found to possess acceptable, physicochemical, mechanical, gelling and mucoadhesion property for delivery of metronidazole.

Development and characterization of mucoadhesive buccal patches of salbutamol sulphate.

Mucoadhesive patch releasing the drug in the oral cavity at predetermined rate may present distinct advantages over traditional dosage forms such as tablets, gels and solutions. The present study was concerned with the preparation and evaluation of mucoadhesive buccal patches for the controlled systemic delivery of Salbutamol sulphate to avoid first pass hepatic metabolism. The developed patches were evaluated for the physicochemical, mechanical and drug release characteristics. The patches showed desired mechanical and physicochemical properties to withstand environment of oral cavity. The in-vitro release study showed that patches could deliver drug to the oral mucosa for a period of 7 h. the patches exhibited adequate stability when tested under accelerated conditions.

Update on nonoxynol-9 as vaginal spermicide.

Nonoxynol-9 (N-9) is a typical surfactant. For more than 30 years that very property of N-9 has been successfully exploited for its spermicidal action. It is available as an over-the-counter, locally acting vaginal spermicide. The suitability of N-9 as a spermicide is elaborated in this article. The reasons why N-9 may fail as a contraceptive are discussed. In spite of many drawbacks, which are mentioned in the article, N-9 is still often resorted to as a locally acting contraceptive. The review ends with suggestions to alter the molecular structure of N-9 and to adjust the dosages.

Sintering of wax for controlling release from pellets.

The purpose of the present study was to investigate incorporation of hydrophobic (ie, waxy) material into pellets using a thermal sintering technique and to evaluate the pellets in vitro for controlled release. Pellets prepared by extrusion-spheronization technology were formulated with a water-soluble drug, microcrystalline cellulose, and carnauba wax. Powdered carnauba wax (4%-20%) prepared by grinding or by emulsification was studied with an attempt to retard the drug release. The inclusion of ground or emulsified carnauba wax did not sustain the release of theophylline for more than 3 hours. Matrix pellets of theophylline prepared with various concentrations of carnauba wax were sintered thermally at various times and temperatures. In vitro drug release profiles indicated an increase in drug release retardation with increasing carnauba wax concentration. Pellets prepared with ground wax showed a higher standard deviation than did those prepared with emulsified wax. There was incomplete release at the end of 12 hours for pellets prepared with 20% ground or emulsified wax. The sintering temperature and duration were optimized to allow for a sustained release lasting at least 12 hours. The optimized temperature and duration were found to be 100 degrees C and 140 seconds, respectively. The sintered pellets had a higher hydrophobicity than did the unsintered pellets. Scanning electron micrographs indicated that the carnauba wax moved internally, thereby increasing the surface area of wax within the pellets.

Differential detection of B. pertussis from B. parapertussis using a polymerase chain reaction (PCR) in presence of SYBR green1 and amplicon melting analysis.

Nucleic acid (DNA) from control stock strains of B. pertussis and B. parapertussis (B. pertussis strain # 9797 and B. parapertussis strain # 15234 from ATCC) was amplified by polymerase chain reaction (PCR) targeting pertussis toxin (PT) promotor region, in presence of SYBR green1 a dye that fluoresces on binding specifically to double stranded DNA; and fluorescent melting profile of amplicon (amplified DNA) was studied. Amplicon of B. pertussis and B. parapertussis generated distinctly different melting bands with melting temperature (Tm) at 89.8 and 91.7 degrees C, respectively. Melting profile and Tm of each randomly selected isolates of B. pertussis and B. parapertussis was identical to that of respective control strains. Distinct difference in Tm between B. pertussis and B. parapertussis specific amplicons allowed differential detection of the two Bordetella species based on a single PCR product. The amplified product of serial diluted control stock of bacteria was analyzed by both agarose gel electrophoresis and melting profile analysis. The analytical sensitivity of detection (1-10 CFU equivalent in the tested volume) by melting profile and Tm analysis was in agreement with that obtained by agarose gel analysis.

Rapid detection of Bordetella pertussis by real-time PCR using SYBR green I and a LightCycler instrument.

A polymerase chain reaction (PCR) assay in real-time for detection of B. pertussis using SYBR green I as the reporter fluorophore and LightCycler instrument (a thermocycler coupled to a fluorescence detection device) was established and evaluated. The amplified amplicon using series diluted control prototype strain (ATCC strain #9797) of B. pertussis was analyzed for the fluorescent melting profile, and melting temperature (Tm) was determined. When examined, amplicons using a representative set of clinical isolates of B. pertussis were found to have the same Tm value (86 +/- 0.5 degrees C, the specificity parameter of detection) as the control prototype strain as expected. Amplified product was also analyzed and detected by agarose gel electrophoresis. The detection limit by fluorescent profile and Tm analysis was 10-fold better than that detected by agarose gel analysis.

A flexible technology for modified release of drugs: multi layered tablets.

Modified release dosage forms offer definite advantages over conventional release formulation of the same drug. Hydrophilic polymers are mainly used for preparation of matrix type controlled delivery systems. The system usually provides nonlinear release profile. The multi-layered matrix system overcomes inherent disadvantages of non-linearity associated with diffusion controlled matrix devices by providing additional release surface with time to compensate for the decreasing release rate. This technology also demonstrates a wide flexibility for various applications. In this article, we review system design, various constructions and formulation parameters of modified release dosage forms.

Clinics in diagnostic imaging (85). Mandible osteoradionecrosis complicated by infection.

Mandibular osteoradionecrosis is a rare but well-known complication that may be seen in patients with head and neck tumours following radiation therapy. A 42-year-old man presented with painful soft tissue swelling and a discharging sinus over the right mandible. Radiographs showed osteolytic destruction. Computed tomography confirmed bony destruction and fragmentation, as well as signs of soft tissue infection. The diagnosis of mandible osteoradionecrosis complicated by infection was confirmed by biopsy and surgically. The clinical and imaging features of osteoradionecrosis are highlighted.

Detection and discrimination of B pertussis and B holmesii by real-time PCR targeting IS481 using a beacon probe and probe-target melting analysis.

A beacon probe was designed to detect one of the two documented single nucleotide changes in IS481 target allele of Bordetella holmesii genome as compared to Bordetella pertussis. PCR amplified product targeting a region of IS481 in presence of the probe was subjected to a post-PCR hybridization and melting cycle. Hybrid of the probe with B. pertussis specific target had a different thermal stability than that with allele having the single nucleotide change in B. holmesii. The melting of B. pertussis-probe hybrid occurred in a single phase; while that of B. holmesii-probe hybrid was biphasic-one for allele identical to that in B. pertussis and the other for that with a single nucleotide change in B. holmesii genome, with a difference in melting temperature (T(m)) of 6.5 degrees C. The characteristic melting profile and T(m) analysis was the basis for discriminatory detection of B. pertussis from B. holmesii. The method was applied in a representative set of clinical isolates of B. pertussis and B. holmesii and the result was in agreement with conventional culture method.

Computed tomographic appearance of colorectal hepatic metastases.

INTRODUCTION: This article reviews the various computed tomography (CT) appearances of hepatic metastases from colorectal primaries and assesses the frequency of occurrence of the various patterns. MATERIALS AND METHODS: This is a retrospective study of the CT appearances of histologically proven colorectal hepatic metastases in a group of 52 patients who had undergone surgical hepatic resection between January 1994 and December 2001. A total of 74 hepatic metastatic lesions were reviewed. All lesions were examined in the portal venous phase. RESULTS: A discernible rim was seen in 54 lesions (73%). Thick rim was present in 36 lesions (48.6%) and thin rim in 18 lesions (24.3%). Enhancement of the rim was present in 62 cases (83.8%). Increased central attenuation was seen in 38 lesions (51.4%). Of these, the centre was heterogeneous in 76.3% and scar-like in 23.7%. A non-enhancing rim was seen in 12 lesions (16.2%) which appeared as lesions with "bevelled edge". Thick enhancing rim with non-enhancing centre was the most common combination in 15 lesions (20.3%). CONCLUSION: An enhancing rim could be seen in 83.8% of lesions. Increased central attenuation was present in 51.4% of the lesions. Familiarity with the various CT appearances may facilitate identification and diagnosis of colorectal liver metastases.

Drug use among the urban poor in Kolkata: behaviour and environment correlates of low HIV infection.

BACKGROUND: HIV infection in injecting drug users (IDUs) has worked as a driving force for further spread of the virus in other population groups. Major metropolitan cities such as Mumbai, Kolkota, Chennai and Delhi have seen a diffusion of injecting drug use within the last decade. The prevalence of HIV infection among injectors ranges from 2% to 30%. Identifying effective interventional elements that have kept the prevalence of HIV low for the past 7 years among IDUs of Kolkata is thus of public health importance. METHODS: A purposive sample of opioid/opiate users was studied. Primary and secondary data on drug users, law-enforcement environment, records at drug treatment centres, jail admission data related to the 'Narcotic Drug and Psychotropic Substance Act' and interventions in other risk groups were collected. Laboratory tests for HIV, hepatitis B surface antigen and syphilis were done on consenting IDUs (n=129) and non-IDUs (n=120). For univariate and multivariate analysis, IDUs were taken as cases and non-IDUs as controls. RESULT: Of the IDUs, 2% were positive for HIV. No non-IDU was HIV-positive. Significantly more non-IDUs (10% v. 4%, p=0.05) were positive for syphilis. Sharing injection equipment within the past 6 months was reported by 71% of IDUs; sharing partners were stable and ranged from 1 to 3. More IDUs compared to non-IDUs reported being in touch with intervention programmes. The police has been tolerant to needle-syringe exchange and oral sublingual buprenorphine substitution conducted in Kolkata. Unlike in the early 1990s, non-IDUs did not switch to injecting during non-availability of brown sugar in the latter half of the 1990s and instead sought tratment. The availability of high quality heroin (>20%-50%) was low and the proportion of moderate quality heroin (>10%-20%) went up during these times due to increased police seizures. No intervention exists in jails despite the fact that a large number of drug users spend time in jail. CONCLUSION: Stable and few injection equipment-sharing partners of IDUs, launching of early targeted interventions among IDUs and sex workers in the city, police tolerance to harm reduction activities and preference of non-IDUs for detoxification during heroin draught periods have kept HIV prevalence at a low level among drug users of Kolkata for the past 7 years. immediate launching of interventions for drug users in jails seems necessary. Similar multi-pronged strategies with targeted and environmental intervention could work in other settings as well.

A modified rapid method of nucleic acid isolation from suspension of matured virus: applied in restriction analysis of DNA from an adenovirus prototype strain and a patient isolate.

This report describes a method for the isolation of nucleic acid from a suspension of matured virus. Nucleic acid (DNA) was isolated from a prototype strain of adenovirus type 7 and a clinical isolate of adenovirus type 7. Instead of the usual method of ultracentifugation, a filtration method was applied to concentrate the virus rapidly and nucleic acid was then isolated by a standard phenol/chloroform/isoamyl-alcohol extraction procedure. The DNA was found to be sufficiently purified to generate a reproducible restriction endonuclease digestion pattern. The clinical isolate of adenovirus type 7 revealed loss of restriction site for the endonuclease HindIII when compared with the prototype strain.

Bordetella pertussis detection by spectrofluorometry using polymerase chain reaction (PCR) and a molecular beacon probe.

Bordetella pertussis was detected by spectrofluorometry following PCR incorporating a molecular beacon probe in the reaction. A DNA fragment from the tandem repeat sequence region (IS 481) of the genome of B. pertussis was amplified in presence of the probe complementary to an internal segment of the amplified DNA fragment. Fluorescein (FAM) and DABCYL were used as the fluorophore and quencher in the probe. The probe was characterized for its signal to noise ratio by homogeneous solution hybridization with a complementary oligonucleotide. Measurement of fluorescent signal at the emission maxima of FAM, immediately after a PCR was used to detect the B. pertussis target, with no additional steps. Presence of B. pertussis in a sample was also examined by agarose gel electrophoresis of the PCR product. A serial diluted stock of B. pertussis (ATCC strain #9797) and fourteen clinical isolates of B. pertussis were examined. The sensitivity of detection by fluorescent measurement was found to be at least in the range of 0.01-0.1 CFU per 10 microl of the sample and was equal to or better than that detected by agarose gel analysis.

The Naval Health Research Center Respiratory Disease Laboratory.

Concern about emerging and reemerging respiratory pathogens prompted the development of a respiratory disease reference laboratory at the Naval Health Research Center. Professionals working in this laboratory have instituted population-based surveillance for pathogens that affect military trainees and responded to threats of increased respiratory disease among high-risk military groups. Capabilities of this laboratory that are unique within the Department of Defense include adenovirus testing by viral shell culture and microneutralization serotyping, influenza culture and hemagglutination inhibition serotyping, and other special testing for Streptococcus pneumoniae, Streptococcus pyogenes, Mycoplasma pneumonia, and Chlamydia pneumoniae. Projected capabilities of this laboratory include more advanced testing for these pathogens and testing for other emerging pathogens, including Bordetella pertussis, Legionella pneumoniae, and Haemophilus influenzae type B. Such capabilities make the laboratory a valuable resource for military public health.