We report a tale of two cases of neonates, where India Ink preparation of cerebrospinal fluid (CSF) received in glass vials was suggestive of Cryptococcus; however, the absence of correlation and report of cases sequentially from the same unit raised alarm bells. Repeat smears using routine in-house and newly prepared stains were prepared and repeat sampling was collected in a strictly sterile specimen container, which resulted in negative microscopy for repeat samples (uncentrifuged and centrifuged). The source of contamination was found to be non-sterile glass vials used for CSF collection. This report underlines the requisite to maintain an aseptic chain from sample collection to processing, to prevent reporting of pseudo-infections that can upset diagnostic accuracy, especially when the aseptic chain has not been maintained.

Cryptococcosis is a fungal infection that may arise in immunocompromised or immunocompetent individuals. This case report seeks to demonstrate the difficulty in diagnosing and treating cryptococcosis based on clinical presentation and radiographic features as together, they mimic other pathological conditions. A 56-year-old female with cirrhosis presented with persistent abdominal pain, dyspnea, vomiting, and diarrhea and was diagnosed with pulmonary cryptococcosis after an initial diagnosis of bacterial pneumonia. With no improvement following antibiotic therapy for suspected bacterial pneumonia, additional imaging was performed with a confirmatory lung biopsy for pulmonary cryptococcosis. The patient initiated antifungal therapy with the anticipation of completing approximately 12 months with follow-up imaging to evaluate improvement. After the patient experienced adverse effects of antifungal therapy and did not achieve significant improvement or recovery in her condition, it was apparent that cryptococcal pneumonia presents both diagnostic and management challenges that must be further explored.

The polysaccharide capsule of Cryptococcus neoformans is the primary virulence factor and one of the most commonly studied aspects of this pathogenic yeast. Capsule size varies widely between strains, has the ability to grow rapidly when introduced to stressful or low-nutrient conditions, and has been positively correlated with strain virulence. For these reasons, the size of the capsule is of great interest to C. neoformans researchers. Inducing the growth of the C. neoformans capsule is used during phenotypic testing to help understand the effects of different treatments on the yeast or size differences between strains. Here, we describe one of the standard methods of capsule induction and detail two accepted methods of staining: (i) India ink, a negative stain, used in conjunction with conventional light microscopy and (ii) co-staining with fluorescent dyes of both the cell wall and capsule followed by confocal microscopy. Finally, we outline how to measure capsule diameter manually and offer a protocol for automated diameter measurement of India ink-stained samples using computational image analysis.

Although rarely used in laparoscopic surgery, computed tomography (CT)-guided marking is useful for targeting small lesions. The present study describes the performance of laparoscopic resection with preoperative CT-guided marking for lateral lymph node recurrence of rectal cancer. A 48-year-old man underwent laparoscopic low anterior resection with D3 lymph node dissection for rectal cancer (postoperative diagnosis, stage IIIb). Postoperative adjuvant chemotherapy was then administered. Solitary lymph node recurrence was observed in the left lateral region after 6 months. Systemic chemotherapy reduced the size of the metastatic lymph nodes; however, 3.5 years after the surgery, the diameter increased by 10 mm, and laparoscopic resection was thus planned. Because the target lesion was small and located deep in the pelvis, preoperative CT-guided marking was performed with India ink injection and embolization microcoil implantation. The markers were clearly identified and the lesion was successfully resected with adequate margins. The approach described in the present study is thus considered potentially useful for the detection of small lesions.

OBJECTIVE: The present study aimed to investigate the accuracy of endoscopic ultrasonography (EUS) combined with Indian ink in locating target vessels of gastric varices (GVs) compared with conventional endoscopic techniques. Additionally, the characteristics of GVs under conventional endoscopy were also explored.
METHODS: All 50 cirrhotic patients with GVs between August 2021 and December 2022 were included in the study. Firstly, conventional endoscopy was employed to identify GVs and to record the expected injection sites. Subsequently, EUS was used to locate the perforated vessel and the injection site was them marked with India ink followed by injection with cyanoacrylate (CYA). Finally, conventional endoscopy was used to examine GVs, to identify the marker points of Indian ink and to compare whether the injection points under conventional endoscopy were consistent with those marked with Indian ink. Furthermore, patients with consistent and inconsistent distribution of endoscopic markers and injection sites were divided into two groups.
RESULTS: EUS could detect the perforating vessels in real time and intuitively. The distribution of markers using EUS was significantly different compared with the injection points obtained by conventional endoscopy (P < 0.001). Therefore, 20 cases were allocated to the consistent group and 30 cases to the non-consistent group. 16 patients who showed red wale signs were obtained in the consistent group and 11 patients in the non-consistent group (P = 0.048). The diameter of the largest GVs was 13.5 (10-15) mm in the consistent group compared with 10 (7.5-10) mm in the non-consistent group (P = 0.006).
CONCLUSIONS: EUS could provide the exact location of GVs, thus more accurately describing the endoscopic characteristics of the GVs. Furthermore, the red wale signs and diameter of the largest GVs obtained using conventional endoscopy were helpful in determining the location of target GVs.

Cryptococcal meningitis represents a severe opportunistic fungal infection primarily observed in individuals with compromised immune systems. It frequently manifests in symptoms like headaches, vomiting, cranial nerve complications, and cognitive alterations. However, it\'s worth noting that up to 15% of cases may exhibit no discernible central nervous system-related symptoms. A 70-year-old male, previously diagnosed with pulmonary tuberculosis and undergoing treatment with anti-tubercular medications, was admitted due to changes in consciousness, sporadic low-grade fever, and cognitive impairment. An in-depth investigation revealed his HIV-negative and non-diabetic status, as well as his preserved immune competence. A plain CT head showed a communicating hydrocephalus and a lumbar puncture was positive for Cryptococcus neoformans. Treatment commenced with an induction regimen encompassing amphotericin and fluconazole, concurrently maintaining the anti-tubercular treatment course. The patient\'s condition displayed improvement, leading to a transition to a maintenance dosage of fluconazole. This case highlighted an extraordinary occurrence of Cryptococcal meningitis in an HIV-negative patient with no history of immunosuppressant use. Notably, Cryptococcal infection should be regarded as a primary consideration in patients afflicted by pulmonary tuberculosis who subsequently present with altered consciousness. The timely identification and proper management of such instances can substantially mitigate the risks of mortality and morbidity associated with this condition.

While upper tract access through the insensate conduit following urinary diversion takes less time and incurs fewer costs than percutaneous kidney access does for the treatment of ureter and kidney pathology, endoscopic ureteroenteric anastomoses (UEA) identification can be difficult. We injected India Ink into the bowel mucosa near the UEA during ileal conduit diversion (IC) to determine the safety and feasibility of ink tattooing. Patients undergoing IC were prospectively randomized to receive ink or normal saline (NS) injections. The injections were placed 1 cm from UEA in a triangular configuration, and loopogram exams and looposcopy were performed to identify reflux (UR), UEA, the tattooing site and strictures in 10 and 11 patients randomized with respect to ink and NS injections, respectively. Ink patients were older (72 vs. 61 years old, p = 0.04) and had a higher Charlson Comorbidity Index (5 vs. 2, p = 0.01). Looposcopy was performed in three ink and four NS patients. Visualization of UEA was achieved in 100% of the ink and 75% of the NS patients (p = 0.26). The ink ureteroenteric anastomotic stricture (UEAS) rate was higher (N = 3 vs. N = 1) and six patients vs. one patients underwent surgery, respectively, for UEAS (p = 0.31). The study was halted early due to safety concerns. Our pilot study demonstrates that ink can be well visualized following injection near UEA during IC. However, the ink cohort had more UEAS than previously cited in the literature and our prior institutional UEAS rate of 6%. While this study sample is small, the higher incidence of UEAS after ink injection led us to question the utility and safety of ink injection following IC.

OBJECTIVE: The purpose of this study is to evaluate the effect of preoperative endoscopic tattooing using India ink (ETI) on the number of retrieved lymph nodes (LNs) dissected during laparoscopic surgery for stage I right-sided colon cancer (RCC).
METHODS: This single-center, retrospective study included stage I RCC patients who underwent laparoscopic surgery between January 2010 and December 2021. The clinicopathological background and number of LNs retrieved were compared between patients managed with and without ETI. A multiple linear regression analysis was used to examine the effect of independent variables on the LN yield.
RESULTS: A total of 169 patients were enrolled. Of these, 89 patients (52.7%) were classified into the ETI group, and 80 (47.3%) were classified into the no-ETI group. There were no significant differences in age, sex, body mass index, or tumor progression between the two groups. A univariate analysis showed that the number of LNs retrieved was significantly higher in female (26 vs. 24, p = 0.026), with tumor localization in the ascending or transverse colon (20 in the cecum, 26 in the ascending colon, 27 in the transverse colon, p < 0.001), and with preoperative ETI (28 vs. 21, p < 0.001). In a multivariate linear regression analysis, female sex (p = 0.0011), D3 lymphadenectomy (p = 0.046), and preoperative ETI (p = 0.012) were independently associated with the LN yield.
CONCLUSIONS: In laparoscopic surgery for stage I RCC, preoperative ETI increased the number of LNs retrieved and allowed for appropriate staging.

Why was India ink used in this case of gingival cancer?Why did the skin change color in this case?What are the benefits and limitations of laser treatment?What is the ideal marking method?

UNASSIGNED: Cryptococcal meningitis is a leading cause of mortality in advanced HIV disease. A positive cerebrospinal fluid cryptococcal antigen (CrAg) test defines cryptococcal meningitis. Herein, we present a patient with serum and cerebrospinal fluid CrAg negative cryptococcal meningitis, despite a positive cerebrospinal fluid India ink examination and quantitative culture.
UNASSIGNED: A 56-year-old HIV-positive Ugandan woman, with an undetectable HIV RNA viral load and CD4+ T-cell count of 766 cells per microlitre presented with signs and symptoms consistent with cryptococcal meningitis. Her serum and cerebrospinal fluid CrAg tests were negative despite having a positive cerebrospinal fluid India ink and quantitative culture. On day 1, she was commenced on intravenous amphotericin B deoxycholate (1mg/kg) for 3 days (considering 10 CFU growth of Cryptococcus spp) in combination with oral flucytosine (100mg/kg) for 7 days and then fluconazole 1200mg once daily for the next 11 days. By day 7, she was symptom free and quantitative cerebrospinal fluid culture was negative for Cryptococcus spp. She was discharged on day 9. At 10 weeks (day +40) and 18 weeks (day +72), she was well and adherent to her antiretroviral therapy and on maintenance phase of cryptococcal meningitis on fluconazole at a dose of 400mg once daily.
UNASSIGNED: This report alerts clinicians managing patients with HIV-associated cryptococcal meningitis to four uncommon clinical scenarios; first, the possibility of negative serum and cerebrospinal fluid CrAg lateral flow assay results in the context of low cerebrospinal fluid fungal burden in a symptomatic patient. Second, possible occurrence of cryptococcal meningitis in a patient with high CD4 T-cell lymphocyte counts. Third, an early seroconversion of cryptococcal antigenaemia following effective fluconazole therapy. Fourth, an early symptomatic relapse of cryptococcal meningitis albeit negative serum CrAg.